Design and Construction of a Lighting System to Illuminate a Photobioreactor A thesis presented to

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Design and Construction of a Lighting System to Illuminate a Photobioreactor A thesis presented to the faculty of the Russ College of Engineering and Technology of Ohio University In partial fulfillment of the requirements for the degree Masters of Science Kyle J. Sink November 2011 © 2011 Kyle J. Sink. All Right Reserved.

Transcript of Design and Construction of a Lighting System to Illuminate a Photobioreactor A thesis presented to

Page 1: Design and Construction of a Lighting System to Illuminate a Photobioreactor A thesis presented to

  

Design and Construction of a Lighting System to Illuminate a 

Photobioreactor 

 

 

 

 

A thesis presented to  

the faculty of  

the Russ College of Engineering and Technology 

of Ohio University 

 

In partial fulfillment  

of the requirements for the degree 

Masters of Science 

 

 

 

Kyle J. Sink 

November 2011 

© 2011 Kyle J. Sink. All Right Reserved.   

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This thesis titled

Design and Construction of a Lighting System to Illuminate a

Photobioreactor

by

KYLE J. SINK

has been approved for

the Department of Mechanical Engineering

and the Russ College of Engineering and Technology by

Greg Kremer

Associate Professor of Mechanical Engineering

Dennis Irwin

Dean, Russ College of Engineering and Technology

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ABSTRACT

SINK, KYLE J., M.S., November 2011, Mechanical Engineering

Design and Construction of a Lighting System to Illuminate a Photobioreactor (103 pp.)

Director of Thesis: Greg Kremer

An internal lighting system for a photobioreactor was designed and constructed

to enhance the growth of green algae. The lighting system is based on a patent for a

solar collector to harness the sun as the primary light source. Fiber optic cables were

fed into the reactor and connected to specially designed light plates that distributed the

light throughout the reactor. The reactor contained Scenedesmus dimorphus, a

microscopic green alga, in a growth media solution. By internally illuminating the

reactor, the energy input cost can be reduced due to the elimination of wasted light.

A 30 liter laboratory scale bubble column reactor was designed and built to test

the objectives laid out. The reactor was designed to be easily assembled and

disassembled, cleaned, and maintained. It was also designed so that the lighting parts

could be exchanged for different configurations if future work necessitates changes.

To design the optimal light plate, the optical simulation software TracePro was

utilized. Information available in literature regarding the absorption and scattering

coefficients of microscopic green algae was analyzed and discussed and the phenomena

of photosynthesis was investigated. A criterion for the selection of the light plate was a

minimum of 20 µEinsteins m-2 s-1 at a distance of 55 cm from the top of the reactor with

a chlorophyll a concentration of 20 mg/L.

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The reactor was inoculated with S. dimorphus and sparged with 1% CO2. The

light intensity was tested using an underwater light sensor in an attempt to validate the

simulation model. Tests also included measurements of pH, chlorophyll a, and turbidity

taken three times daily to study the growth potential of the reactor. The results of the

tests show that the algae grows steadily in the reactor indicating that the proposed

design could be an effective method for growing algae as a petroleum substitute and for

the biologic sequestration of CO2.

Approved:

Greg Kremer

Associate Professor of Mechanical Engineering

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TABLE OF CONTENTS

Page

ABSTRACT ......................................................................................................................................... 3

LIST OF FIGURES ............................................................................................................................... 7

LIST OF TABLES ................................................................................................................................. 9

1 INTRODUCTION ........................................................................................................................... 10

1.1 Energy Demands and Costs Predictions ............................................................................... 10

1.2 Biomass ................................................................................................................................ 12

1.3 Biofuels ................................................................................................................................ 13

1.4 Algae .................................................................................................................................... 14

1.5 Photobioreactors ................................................................................................................. 16

1.6 Project significance .............................................................................................................. 19

1.7 Objectives ............................................................................................................................ 19

2 LITERATURE REVIEW ................................................................................................................... 21

2.1 Carbon Sequestration .......................................................................................................... 21

2.2 Photobioreactors ................................................................................................................. 22

2.3 Bubble Column Reactor ....................................................................................................... 26

2.3.1 Gas Velocity ................................................................................................................... 28

2.3.2 Gas Holdup .................................................................................................................... 28

2.4 Light and dark cycles ............................................................................................................ 29

2.5 Compensation Point............................................................................................................. 34

2.6 Light Transmittance ............................................................................................................. 35

3 DESIGN APPROACH ..................................................................................................................... 38

3.1 Design Parameters and Constraints ..................................................................................... 38

3.1.1 Fiber Optics ................................................................................................................... 38

3.1.2 Gas Sparger ................................................................................................................... 40

3.2 Algae Specimen .................................................................................................................... 40

3.3 Optical Simulations .............................................................................................................. 41

3.3.1 Simulation Software ...................................................................................................... 42

3.3.2 Simulation material properties ..................................................................................... 43

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3.3.3 Light plate design and testing ....................................................................................... 50

3.3.4 Benefits of light plates .................................................................................................. 53

3.5 Illuminator Cooling ............................................................................................................... 64

3.6 Reflective Coating ................................................................................................................ 64

3.7 Test Plan ............................................................................................................................... 67

3.7.1 Analytical Procedures for fluid properties .................................................................... 67

3.7.2 Light Measurements ..................................................................................................... 69

4 RESULTS AND DISCUSSION .......................................................................................................... 73

4.1 Simulation Results ................................................................................................................ 73

4.2 Reactor results ..................................................................................................................... 78

4.3 Scattering due to bubbles .................................................................................................... 90

4.4 Circulation ............................................................................................................................ 91

5 CONCLUSIONS ............................................................................................................................. 94

5.1 Working System ................................................................................................................... 94

5.3 Light Plate Design ................................................................................................................. 95

6 RECOMMENDATION AND FUTURE WORK .................................................................................. 97

6.1 Recommendations ............................................................................................................... 97

6.1.1 Lighting system ............................................................................................................. 97

6.1.2 Reactor components ................................................................................................... 100

6.2 Future Work ....................................................................................................................... 102

WORK CITED ................................................................................................................................. 105

APPENDIX A: Growth material ..................................................................................................... 110

APPENDIX B: Additional plots ...................................................................................................... 111

APPENDIX C: Cost ......................................................................................................................... 113

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LIST OF FIGURES

FIGURE 1. CO2 EMISSIONS OVER TIME FROM COAL FROM 1980 TO 2008 (U.S. ENERGY INFORMATION

ADMINISTRATION, 2010) ............................................................................................................ 11

FIGURE 2. CONVERSION PROCESS FOR BIOMASS TO BIOFUEL (MCKENDRY, 2002) ........................................ 14

FIGURE 3. MICROSCOPIC GREEN ALGAE (WILSON, 2009), LEFT, AND GIANT ALGAE IN NEW ZEALAND (ORIZARSKA,

2007) ...................................................................................................................................... 15

FIGURE 4. ABANDONED HORIZONTAL TUBE FACILITY (MIRON A. S., 1998) .................................................. 17

FIGURE 5. VARIOUS BIOREACTOR SETUPS, THE LOOP REACTOR LABELED HERE IS THE AIRLIFT REFERRED TO IN THIS

WORK (CHISTI, 1983) ................................................................................................................. 18

FIGURE 6. GEOLOGICAL AND BIOLOGICAL SEQUESTRATION METHODS (PLUNGING INTO CARBON SEQUESTRATION

RESEARCH, 2000) ...................................................................................................................... 22

FIGURE 7. NEAR HORIZONTAL TUBE REACTOR (CARVALHO, MEIRELES, & MALCATA, 2006) ........................... 25

FIGURE 8. POSSIBLE FLOWS IN A BUBBLE COLUMN (KANTARCI, BORAK, & ULGEN, 2005) .............................. 27

FIGURE 9. COLUMN DIAMETER AND GAS VELOCITY EFFECTS ON BUBBLE FLOW (KANTARCI, BORAK, & ULGEN,

2005) ...................................................................................................................................... 28

FIGURE 10. COMPARISON OF GROWTH RATES AND CHLOROPHYLL A CONCENTRATIONS WITH RESPECT TO

DIFFERENT LIGHT CYCLE PERIODS (CARR & WHITTON, 1982) ............................................................. 30

FIGURE 11. CELL GROWTH OVER TIME WHEN EXPOSED TO CONSTANT LIGHT FOR APHANOTHECE MICROSCOPICA

(JACOB-LOPES, 2009) ................................................................................................................. 31

FIGURE 12. CELL CONCENTRATIONS WITH DIFFERENT DARK:LIGHT CYCLES (JACOB-LOPES, 2009) .................... 32

FIGURE 13. CO2 ABSORPTION AS COMPARED TO A 24 HOUR PERIOD OF LIGHT (JACOB-LOPES, 2009) .............. 33

FIGURE 14. LIGHT CURVE COMPARING LIGHT INTENSITY TO PHOTOSYNTHESIS RATES (GRAHAM & WILCOX, 2000)

............................................................................................................................................... 35

FIGURE 15. LIGHT BEING REFLECTED AT MEDIA INTERFACE ......................................................................... 35

FIGURE 16. LIGHT REFRACTING (WEISSTEIN, 2007) ................................................................................. 36

FIGURE 17. FIBER OPTIC ILLUMINATOR ................................................................................................... 39

FIGURE 18. FIBER OPTIC POLISHER......................................................................................................... 39

FIGURE 19. SCENEDESMUS DIMORPHUS IN SOLUTION ............................................................................... 41

FIGURE 20. LIGHT ATTENUATION CALCULATIONS PROCESS (BASS, 2010) ..................................................... 45

FIGURE 21. SCATTERING COEFFICIENT CURVES BASED ON DATA FROM TABLE 5 ............................................. 48

FIGURE 22. SHORT (LEFT) AND LONG (RIGHT) VERSIONS OF THE LIGHT PLATE (UNITS IN INCHES) ...................... 51

FIGURE 23. LIGHT PATHS THROUGH THE UNIQUE PLATE DESIGN ................................................................. 53

FIGURE 24. 35° LIGHT PLATE TO BE INSTALLED IN BCR ............................................................................. 56

FIGURE 25. BOTTOM PLATE HOLDER ...................................................................................................... 56

FIGURE 26. TOP PLATE HOLDER ............................................................................................................ 57

FIGURE 27. PARTIALLY CONSTRUCTED COMPONENTS OF BCR .................................................................... 58

FIGURE 28. SLOT MECHANISM FOR LOWER DISK ...................................................................................... 59

FIGURE 29. SLOT MECHANISM FOR UPPER DISK ....................................................................................... 59

FIGURE 30. GAS DELIVERY SYSTEM ........................................................................................................ 60

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FIGURE 31. GAS DELIVERY SYSTEM (LOOKING FROM UNDER THE REACTOR) .................................................. 61

FIGURE 32. INTERNAL LIGHT COMPONENTS PRIOR TO ASSEMBLY WITH COLUMN ........................................... 62

FIGURE 33. LIGHTING COMPONENTS INSTALLED IN COLUMN ...................................................................... 63

FIGURE 34: COOLING SYSTEM FOR ILLUMINATOR ..................................................................................... 64

FIGURE 35. BCR COVERED IN ALUMINUM FOIL ........................................................................................ 65

FIGURE 36. LIGHT PLATES COVERED IN ALUMINUM FOIL EXCEPT OVER THE ANGLED FACES .............................. 66

FIGURE 37. LIGHT SENSOR FIXTURE INCLUDING DEPTH ROD ....................................................................... 69

FIGURE 38. SENSOR ORIENTATION INSIDE REACTOR FOR ACCURATE LIGHT MEASUREMENTS ............................ 70

FIGURE 39. DEPTH ROD WITH MARKINGS SHOWN .................................................................................... 71

FIGURE 40. DISTANCES BETWEEN VARIOUS COMPONENTS OF THE REACTOR TO UNDERSTAND PLACEMENT OF

SENSOR USING DEPTH ROD MARKING. THE SENSOR IN THIS CASE IS AT A DEPTH OF 24 CM. ILLUSTRATION IS

NOT SCALE. ................................................................................................................................ 71

FIGURE 41. RESULTS OF LIGHT SIMULATIONS FOR TWO DIFFERENT CHLOROPHYLL A CONCENTRATIONS

THROUGHOUT THE REACTOR. (A) 35 PLATE, (B) 35 LONG PLATE, (C) 45 PLATE, (D) 45 LONG PLATE ......... 75

FIGURE 42. RESULTS OF LIGHT SIMULATIONS FOR TWO DIFFERENT CHLOROPHYLL A CONCENTRATIONS

THROUGHOUT THE REACTOR. (A) 55 PLATE, (B) 55 LONG PLATE, (C) 60 PLATE, (D) 60 LONG PLATE ......... 76

FIGURE 43. LIGHT INTENSITY DISTRIBUTION (LEFT) AND COLOR MAP (RIGHT) OF THE 35° LONG PLATE AT A DEPTH

OF 425 MM ............................................................................................................................... 77

FIGURE 44. LI-192 UNDERWATER LIGHT SENSOR (LEFT) AND LI-190 LIGHT SENSOR (RIGHT) .......................... 83

FIGURE 45. COMPARISON OF CALIBRATED AND UNCALIBRATED LIGHT SENSOR AT A DISTANCE OF 1.5" FROM LIGHT

SOURCE IN AIR ............................................................................................................................ 84

FIGURE 46. COMPARISON OF CALIBRATED AND UNCALIBRATED LIGHT SENSOR AT A DISTANCE OF 4" FROM LIGHT

SOURCE IN AIR ............................................................................................................................ 84

FIGURE 47. MELTED FIBER OPTIC CABLE AFTER PROLONGED USE WITH THE ILLUMINATOR ............................... 86

FIGURE 48. ABSORPTION COEFFICIENTS FOR WATER CONTAINING PHYTOPLANKTON FOR CHLOROPHYLL A

CONCENTRATION FROM 0.2 AND 18.4 MG M-3 (PRIEUR & SATHYENDRANATH, 1981) ......................... 87

FIGURE 49. CHLOROPHYLL A AND TURBIDITY CHANGES OVER COURSE OF EXPERIMENT. INOCULATION OCCURRED

AT 7 P.M. ON DAY 0 WITH THE FIRST MEASUREMENTS TAKEN ON DAY 1. ............................................. 88

FIGURE 50. DIFFERENCE IN LIGHT PENETRATION WITH RESPECT TO DISTANCE BASED ON BUBBLING. THE POINTS

ARE THE DAY AND TIME OF THE MEASUREMENT, I.E. 1-10 WAS TAKEN AT 10:00 ON THE DAY ONE OF

TESTING. ................................................................................................................................... 91

FIGURE 51. PROGRESSION OF MIXING VIA A DYE TEST ............................................................................... 93

FIGURE 52. LI-193 SPHERICAL LIGHT SENSOR (UNDERWATER PAR MEASUREMENT, 2011) .......................... 98

FIGURE 53. METAL HALIDE FIBER OPTIC ILLUMINATOR (WIEDAMARK, 2011) ............................................... 99

FIGURE 54. LIGHT LOSS OF FROM COUPLER AT INTERFACE WITH LIGHT PLATE .............................................. 101

FIGURE 55. ALTERNATIVE METHOD OF INTERFACING FIBERS WITH LIGHT PLATES BY EMBEDDING FIBERS INTO THE

PLATES .................................................................................................................................... 101

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LIST OF TABLES

TABLE 1. TYPICAL SOURCES OF BIOMASS FOR ENERGY PRODUCTION (STAM, 2009) ....................................... 13

TABLE 2. COMPARISON OF OPEN AND CLOSED ALGAE GROWTH SYSTEMS (UNITED NATIONS, 2009) ................ 23

TABLE 3. COMPARISON OF SEVERAL REACTOR TYPES (CARVALHO, MEIRELES, & MALCATA, 2006) .................. 24

TABLE 4. CHLOROPHYLL A SPECIFIC ABSORPTION COEFFICIENTS WITH RESPECT TO WAVELENGTH (BASS, 2010) .. 46

TABLE 5. SCATTERING COEFFICIENTS (BW) FOR WATER AND SEA WATER (BASS, 2010)................................... 47

TABLE 6. SCATTERING COEFFICIENT WITH RESPECT TO WAVELENGTH FOR PURE SEA WATER ............................. 48

TABLE 7. SCATTERING COEFFICIENTS WITH RESPECT TO WAVELENGTH FOR PURE WATER ................................. 49

TABLE 8. TWO VARIABLE ANALYSIS OF LIGHT INTENSITY USING BEER-LAMBERT'S LAW .................................... 50

TABLE 9. RESULTS FROM 8/16/2011 .................................................................................................... 79

TABLE 10. RESULTS FROM 8/17/2011 .................................................................................................. 79

TABLE 11. RESULTS FROM 8/18/2011 .................................................................................................. 80

TABLE 12. RESULTS FROM 8/19/2011 .................................................................................................. 80

TABLE 13. RESULTS FROM 8/20/2011 .................................................................................................. 81

TABLE 14. RESULTS FROM 8/21/2011 .................................................................................................. 81

TABLE 15. RESULTS FROM 8/22/2011 .................................................................................................. 82

TABLE 16. DIFFERENCE IN MEASUREMENTS WHEN SAMPLES ARE TAKEN FROM THE TOP AND BOTTOM OF THE

REACTOR ................................................................................................................................... 89

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1 INTRODUCTION

1.1 Energy Demands and Costs Predictions

Power generation is a necessary element of today’s society. However, society is

no longer as accepting of the methods used to generate the energy we crave. The older

methods are becoming too expensive and are seen as too great a burden on the

environment. In the United States in 2009 alone energy consumption was 94.87

quadrillion Btu [1]. Although due to economic recession this value is down from 2008

and 2007 when consumption was 99.38 and 101.55 quadrillion Btu, respectively, the EIA

predicts that the energy demand by the United States will steadily increase at an

average rate of .2% a year until 2030 under current trends. While this trend will present

challenges by itself, on the global scale developing countries are quickly taking a larger

share of the global energy supply.

Asia is expected to see the biggest increase, with China leading the way. It is

projected that there will be a 3% yearly increase in energy consumption by Asia alone,

potentially topping 240 quadrillion Btu in 2030 up from just 47.4 quadrillion Btu in 1990.

A great majority of this energy is expected to come from coal. There will be similar

increases in South America where energy demands could rise from 149.9 quadrillion Btu

in 1990 to 400 quadrillion Btu in 2030, again with the biggest increase coming from coal.

This increase in the consumption of coal will lead to an increase in CO2 output.

According to the EIA Carbon Dioxide Emissions report the worldwide output of CO2 was

12,897.9 million tons in 2008, compared to 6,572.5 million tons in 1980. Figure 1 shows

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the emission trends for the last 30 years. This huge spike in emissions is expected to

keep growing due to the industrialization of developing countries where they lack

emissions standards that are in place in the developed world.

This rapidly increasing consumption raises two concerns. The first is that coal,

and all fossil fuels, are nonrenewable. This means that there is a limited supply and that

this supply will become harder to reach and therefore increase in price as the end draws

nearer. The other concern is that CO2 is a greenhouse gas. Greenhouse gases retain

heat that would normally bounce off the surface of the earth and into space. While no

proven effects of this have been observed to be caused by human emissions, it is best to

prevent putting excessive amounts of anything into the air. A method of sequestration

must be developed which is a focus of this work.

Figure 1. CO2 emissions over time from coal from 1980 to 2008 (U.S. Energy Information

Administration, 2010)

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1975 1985 1995 2005 2015

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1.2 Biomass

The Biomass Energy Center in the United Kingdom describes biomass as the

“biological material derived from living, or recently living organisms”, generally used

when talking about plant material (Biomass Energy Centre, 2008). Biomass is carbon

based, but does contain organic compounds and elements like oxygen and nitrogen as

well as some other heavy metals. Fossil fuels are concentrated forms of biomass; they

are just not considered “recent” biomass. For the context of this paper biomass will

refer to new matter on a human times scale while fossil fuels will refer to old matter

based on a geological time scale. Biomass is considered a carbon neutral fuel because

when plant material is burned it releases CO2 back into the atmosphere. This CO2 is then

used by plants for growth. This is called the carbon cycle. When biomass is burned and

new plant material is planted and allowed to grow the total carbon in the atmosphere is

relatively unchanged. It is theorized that fossil fuels require millions of years to form;

therefore when fossil fuels are burned, CO2 is released that has been stored for millions

of years and will add to the total carbon in the atmosphere in its current state.

Biomass comes in many forms, from waste material to crops grown specifically

for energy; Table 1 shows many traditional sources of biomass. However this creates a

problem of supply. Quick access to large quantities of plant biomass can be difficult as

only a specific amount is grown or collected. Once this has been used more must be

grown, and for traditional forms of biomass this could take a full season. It is therefore

vital that a new source be found that can be grown quickly and in large amounts. That

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source is believed to be algae; however the process of growing algae still has not been

perfected on an industrial scale.

Table 1. Typical sources of biomass for energy production (Stam, 2009)

1.3 Biofuels

Biomass can be converted to biofuels using natural chemical processes. The

process of anaerobic digestion uses bacteria to break down the biomass in a slurry into

a biogas similar to natural gas. Fermentation is generally done on cereal or sugar plants

and is used to produce ethanol, the process is the same used to make alcoholic drinks.

Composting is similar to anaerobic digestion except that it is typically a dry process.

Figure 2 outlines various processes and results from different conversions of biomass.

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Figure 2. Conversion Process for Biomass to Biofuel (McKendry, 2002)

These methods of producing a renewable source of fuel are very promising

except that they require tremendous amounts of crops that at the present are not

sustainable along with maintaining current food supplies. An example of the

unsustainability of terrestrial biomass, such as sugar or corn, for fuel is that to grow

enough biomass for 1 liter of fuel requires 10,000 L of water (Wijffels & Barbosa, 2010).

However, the use of algae is becoming a popular alternative to traditional biomass

supplies as algae can be grown quickly and with relatively little input. To grow

approximately 1 kg of algal mass requires only .75 L of water for photosynthesis (Wijffels

& Barbosa, 2010).

1.4 Algae

Algae is a broad term that represents a vast grouping of organisms that can

range in size from microscopic to enormous seaweeds. Nearly all algae contain

Chlorophyll a and β-carotene for photosynthesis, and these types of algae are the main

focus of this work. Figure 3 show various types of algae.

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Figure 3. Microscopic green algae (Wilson, 2009), left, and giant algae in New Zealand

(Orizarska, 2007)

Algae produce a large amount of the oxygen in the atmosphere. Algae also serve

as the basis of the earth’s food chain because they are rich in hydrocarbons. Algae were

first looked at as a potential food source that was high in nutrients and quick to grow in

the 1950’s, though they are currently used to make products ranging from cosmetics to

pharmaceuticals to dyes. Some of these products cannot be made any other way.

Research for algae based biofuels intensified in the 1970’s with the energy crisis, along

with other plans for renewable energy (United Nations, 2009).

Along with photosynthesis algae also perform respiration, like other living

creatures, in which oxygen is needed; carbon dioxide is only consumed for

photosynthesis. Respiration occurs constantly, however, it is most noticeable when

photosynthesis is not occurring, such as during dark cycles. During dark cycles, algae

will use some of the growth to sustain itself so there will be a slight loss during these

times. During a full night up to 25% of growth from the previous day can be used,

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depending on temperature, for respiration. Constant lighting has been used to counter

this; however using dark/light ratios to control loss can also be used. Photosynthesis

utilizes less than 50% of the available light spectrum, and it does so very inefficiently.

Algae converts only 11% of light energy to chemical (United Nations, 2009).

1.5 Photobioreactors

Processes that involve microorganisms require reconfiguration of the multiphase

reactor into a bioreactor. A bioreactor must be capable of delivering nutrients to and

removing waste generated by the living organisms. Processes involving bioreactors

include production of penicillin to various enzymes to food products like vinegar, yogurt,

cheese, and beer (Chisti, 1983). These processes do not require light and therefore are

done in the dark. However, new applications of these systems make use of

photosynthetic organisms so the introduction of lighting systems is vital.

There are two types of reactors for growing algae, pond systems and closed

photobioreactors. Both of these systems have distinct advantages and disadvantages.

The major drawbacks to the open systems are the land usage, difficulty in diffusion of

nutrients and CO2, and controlling contamination. Putting nutrients into the slurry can

be a very slow and inefficient process.

Internally lit bioreactors can be made in the form of horizontal tubes or vertical

columns. The horizontal tube systems have been introduced first at a production scale;

however it has proven very difficult to remove excess oxygen from the system.

Nutrients and CO2 are put into the system at one point and waste is removed at the end

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of the tubes. This means that nutrients are usually depleted long before they reach the

end of the tube. The excess oxygen from respiration displaces the CO2 and causes the

algae to starve because it cannot perform photosynthesis. This effect was increased

because mixing is difficult in long horizontal tubes. Tubular systems also require a lot of

land, are difficult and expensive to build and maintain, and have never worked as

theoretically planned, for example the Photobioreactor Ltd facility in Spain was

abandoned because it did not produce as planned. Figure 4 shows a picture of the

tubular structure that was abandoned.

Figure 4. Abandoned horizontal tube facility (Miron A. S., 1998)

The bioreactors that have shown to be the best for reactions requiring light have

been the various vertical column reactors. Typically these kinds of reactors have very

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high gas hold up values and with the bubble column specifically there is a significant

advantage with respect to the light/dark cycles than the horizontal tube reactors (Miron

A. S., 1999). Figure 5 shows some of the variations available with the bubble column.

The bubble column is superior because there is a constant flow of nutrients and waste

material. CO2 is pumped into the bottom of the reactor and allowed to bubble to the

top, as it does so it is consumed by the algae. Also, as O2 is produced it will rise to the

top and will exit the system in an exhaust opening. This bubbling also generates a

mixing current in the slurry which is vital to the algae survival and growth as it

continually brings the algae cells individually into contact with CO2.

Figure 5. Various bioreactor setups, the loop reactor labeled here is the airlift referred

to in this work (Chisti, 1983)

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1.6 Project significance

The Institute for Sustainable Energy and the Environment (ISEE) has been

studying ways to more effectively use non-renewable energy sources and developing

new renewable sources to take the center role of meeting our energy demands. ISEE

has built and tested an airlift style photobioreactor and has need of a bubble column

reactor to do side by side comparisons for algal growth efficiencies. With the bubble

column reactor, future work can continue in the area of carbon sequestration and

petroleum substitutes. This work will also add to the literature information about the

use of lighting in a bubble column reactor, which at the present is greatly lacking.

1.7 Objectives

The main objective of this thesis was to design and construct a lighting system

that will efficiently introduce light into a bioreactor to create conditions under which

algae can grow. To achieve this, a secondary objective was to create a working bubble

column style reactor. This reactor will allow more in depth study of light conditions and

other growth parameters. It is not the objective of this work to achieve a specific

production level for algae growth. Past work was studied on maximizing light diffusion

and penetration through aqueous solutions. A literature review was done to

understand bioreactor design, culturing algae, and the use of internal lighting and how it

can be applied in this situation. The bubble column is comparable in volume to the

airlift system already constructed so that future work can be done on a side by side

comparison as to which design is the most efficient.

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To achieve these objectives, optical simulations were performed to determine

the most efficient light guide design. The light guides were constructed based on the

results and tested in a reactor similar to the simulation model. The light guides were

designed to be capable of producing diffuse light at an intensity value above the

compensation point for green algae. While no specific growth rate will be attempted,

the reactor must show that algae growth is supported.

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2 LITERATURE REVIEW

2.1 Carbon Sequestration

From Figure 1 in the Introduction the trends of CO2 output can be clearly seen to

be increasing. CO2 has been reported to encourage global climate change, and

therefore increasingly stringent restrictions have been placed on burning fossil fuels.

One method to adhere to these restrictions is to sequester, or isolate and store, the CO2

using a means that will not contribute to an increase in atmospheric levels.

Sequestration can take one of two forms; geological or biological. Geological

sequestration involves pumping CO2 into used natural gas pockets or some other empty

area underground. Biological sequestration is using photosynthetic organisms to

remove CO2 from the air. Figure 6 shows both methods. Biological sequestration does

not permanently remove CO2 from the air but instead is intended to “trap” it in a cycle

of continual absorption for photosynthetic use, release by combustion, and then again

absorbed (Kumar, 2010).

However, new research has shown that a more efficient method of biological

sequestration is using algae. Algae sequester CO2 in three different ways: from the air,

from industrial exhaust, or in solid carbonate states (Wang, 2008). Algae naturally take

CO2 from air like all photosynthetic organisms; however air is typically only made up of

0.03-0.06% CO2, which place a limit on algae growth. Industrial flue gas contains

approximately 15% CO2, and can be utilized at little to no additional cost. This source

could be fed directly to the algae or it can be chemically changed to a carbonate and fed

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to algae. A total of 1.8 tons of CO2 are required to grow one ton of algal biomass

(Wijffels & Barbosa, 2010).

Figure 6. Geological and biological sequestration methods (Plunging into Carbon

Sequestration Research, 2000)

2.2 Photobioreactors

The first reactors for growing algae were open ponds systems, both natural and

manmade. Algae from the ocean have been harvested for food when it washes on

shore. These systems relied on solar energy for photosynthesis and for small scale

needs can be practical. However there are some stark disadvantages to this type of

system. In a report from the United Nations open ponds and closed photobioreactors

were compared.

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Table 2. Comparison of open and closed algae growth systems (United Nations, 2009)

Parameter of Issue Open ponds and raceways Photobioreactor (PBR)

Required space High For PBR itself, low

Water loss Very high, may also cause salt

precipitation

Low

CO2-loss High, depending on pond depth Low

Oxygen concentration Usually low enough because of

continuous spontaneous

outgassing

Build-up in closed system requires gas exchange

device (O2 must be removed to prevent inhibition

of photosynthesis and photo-oxidative damage)

Temperature Highly variable, some control

possible by pond depth

Cooling often required (by spraying water on PBR

or immersing tubes in cooling baths.

Shear Usually low (gentle mixing) Usually high (fast and turbulent flows required for

good mixing, pumping through gas exchange

devices

Cleaning No issue Required (wall growth and dirt reduce light

intensity), but causes abrasion, limiting PBR life-

time.

Contamination risk High (limiting the number of

species can be grown)

Low (medium to low)

Biomass quality Variable Reproducible

Biomass concentration Low, between 0.1 and 0.5 g/l High, generally between 0.5 and 8 g/l

Production flexibility Only a few species possible,

difficult to switch

High, switching possible

Process control and

reproducibility

Limited (flow speed, mixing,

temperature only by pond depth)

Possible within certain tolerances

Weather dependence High (light intensity, temperature,

rainfall

Medium (light intensity, cooling required)

Start-up 6-8 weeks 2-4 weeks

Capital costs High ($100,000 per hectare) Very high ($250,000 to $1,000,000 per hectare,

PBR and supporting systems)

Operating costs Low (stirring mechanism, CO2

addition

Higher (CO2 addition, oxygen removal, cooling,

cleaning, maintenance)

Harvesting cost High, species dependent Lower due to high biomass concentration and

better control over species and conditions

Current commercial

applications

5,000 (8 to 10,000) t of algal

biomass per year

Limited to processes for high assed value

compounds or algae used in food and cosmetics

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Photobioreactors (PBRs) differ from open pond systems in that the necessary

conditions for growth, temperature, pH, nutrient supply, etc., can be strictly controlled

and regulated. PBR’s currently rely on solar energy radiating from the exterior of the

reactor for photosynthesis, but this will be required to change as scale up occurs. As

volumes increase external lighting will no longer be capable of supplying energy to all

algae cells as light is absorbed quickly upon entering an algae slurry.

Most PBR’s can be broken down into three categories: tubular, flat plate, and

fermenter reactors (Carvalho, Meireles, & Malcata, 2006). Tubular and flat plate

designs are used most frequently because natural sunlight is used, which has very low

costs associated with it. Fermenters require some kind of internal illumination. Carvalho

provided a comparison of several reactor designs in Table 3. Potential productivity is

also given for various algae species. From looking at this table the fermenter type is

clearly not preferred as it has a poor light harvesting efficiency, difficult scale up, and

very low productivity.

Table 3. Comparison of several reactor types (Carvalho, Meireles, & Malcata, 2006)

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Tubular reactors can be vertical, horizontal, or helical, with most being the first

two. Vertical tubular reactors are usually in the form of airlift reactors or bubble

columns, which is the focus of this work and will be discussed in detail later. Horizontal

tubular reactors pump the algae mixture through a long series of tubes. Some success

has been achieved with this method, however, a major problem of heat generation

arises due to the pumps. Figure 7 shows a generic design for a horizontal tube system.

Figure 7. Near horizontal tube reactor (Carvalho, Meireles, & Malcata, 2006)

Flat plate reactors are narrow panels designed to efficiently use as much

external light as possible. There are several shapes that have been developed and can

be pumped or bubbled. Fermenters have low area-to-volume ratios and therefore low

sunlight harvesting efficiencies. Internally lit fermenters have been developed on a

bench scale, however until an effective method of scale to production is found,

fermenters will not be used for photosynthetic processes.

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2.3 Bubble Column Reactor

Bubble column reactors (BCR) are generally a transparent cylindrical tube with a gas

sparger at the bottom. Gas is pumped into the column through the sparger and is

bubbled up through the system which contains a liquid-solid mixture, called a slurry.

This bubbling provides necessary nutrients to the system as well as mixes the system.

BCR’s have several advantages over other reactor types (Kantarci, Borak, & Ulgen,

2005):

• Excellent heat and mass transfer characteristics

• Low maintenance and operating costs

• Few moving parts

• Active system catalyst addition and removal

• High durability

Bubble columns are not currently being utilized as PBR’s with external lighting

because the column diameter cannot exceed 0.2 m without a severe loss of light

intensity (Miron A. S., 2000). If a suitable scale up could be found to incorporate

internal lighting, the bubble column could be a major alternative to common reactors

today.

There are several flows that are possible with the bubble column; different flows are

useful in different situations. These flows are shown in Figure 8. The homogeneous

flows occur at low gas velocities, typically less than 5 cm/s. A homogeneous flow

consists of uniform bubble sizes that rise at a uniform rate and generate only gentle

mixing. Heterogeneous flows usually occur when gas velocity exceeds 5 cm/s and is

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made up of big and small bubbles. The increase of energy creates more chaotic mixing

and circulation. It has been observed however that the gas-liquid mass transfer is lower

in a heterogeneous flow than homogenous (Kantarci, Borak, & Ulgen, 2005). Turbulent

flow can also introduce additional shear stresses, which may be unacceptable for

organisms, as compared to homogeneous flows. In bubble columns the only sources of

stress are from the bubble movement, which is dependent on gas velocity, and any

lighting structures (Perez, 2006). Flow patterns depend on column diameter and gas

velocity and can be estimated using Figure 9.

The most important parameters to bubble column design are gas holdup, bubble

characteristics, and mass transfer. Gas holdup is the fractional percentage of gas in the

liquid and will be discussed more below. Bubble characteristics are determined by

liquid and gas properties, sparger pore size, superficial gas velocity, and diameter of the

reactor.

Figure 8. Possible flows in a bubble column (Kantarci, Borak, & Ulgen, 2005)

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Figure 9. Column diameter and gas velocity effects on bubble flow (Kantarci, Borak, &

Ulgen, 2005)

2.3.1 Gas Velocity

Superficial gas velocity is the average velocity of gas being bubbled into the

bubble column through a sparger, which is given by Equation 1.

�� � ��

Where Ug = Superficial gas velocity

Q = Volumetric flow rate

A = Cross sectional area

(1)

2.3.2 Gas Holdup

One of the most important design criteria for the bubble column is the gas

holdup value. Gas holdup is defined as the volume fraction of gas that is taken up by

gas bubbles and is given by Equation 2 (Miron A. S., 1999).

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ε � 3.317 × 10�� �P�V���.��

Where � � ���ℎ�� !"

#$%& � '"()*+*)"�,(-*."!/

(2)

The specific power input is the power input divided by the volume of liquid in the

reactor. The power input can be adjusted by changing the rate at which gas is bubbled

into the system. The specific power input can also be defined by equation 3.

P�V� �ρ�gU�

(3)

Where ρL is the density of the liquid phase, g is the gravitational constant, and UG

is the superficial gas velocity as defined earlier. Gas holdup has been shown to increase

as gas velocity and pressure increase and decrease when liquid velocity increases. Gas

holdup is also affected by the column height when the height to diameter ratio of the

column is less than 5:1. When the ratio is larger than this the height effect on holdup is

negligible (Kantarci, Borak, & Ulgen, 2005).

2.4 Light and dark cycles

Photosynthesis is a chemical process that occurs in organisms containing

chlorophyll: terrestrial plants, algae, and some bacteria. These organisms can harness

the energy in a photon to create sugars that they feed on. While light is required for

photosynthetic organisms to survive, a dark cycle is also needed. This dark cycle is

commonly called the Calvin cycle and is when the cell actually uses the photon that it

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captures during the light cycle. During the Calvin cycle the cell goes through the process

of respiration like every creature, in which it consumes oxygen and part of the biomass

to replace chemicals and repair cell structure damaged during photosynthesis.

The Calvin cycle will continue until light is available again so the longer the cell is

in the dark the more mass loss will occur. To counter this many have tried to provide

constant light, however research has shown that constant light does not provide a clear

advantage to intermittent light (Carr & Whitton, 1982; Jacob-Lopes, 2009). Figure 10

shows the relationship between light periods and the average growth rate. For this

experiment the total period is 24 hours so on the right is 24 hours of light and the left is

24 hours of dark. As the process approaches 24 hours of light the effects of the

additional light become less important. This experiment was done with Oscillatoria

agardihii.

Figure 10. Comparison of growth rates and chlorophyll a concentrations with respect to

different light cycle periods (Carr & Whitton, 1982)

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A similar experiment was done by using Aphanothece microscopica with varying

dark/light ratios. Figure 11 shows the cell concentration over time using constant

lighting conditions.

Figure 11. Cell growth over time when exposed to constant light for Aphanothece

microscopica (Jacob-Lopes, 2009)

With constant light the alga reaches a steady state of approximately 5000 mg/L

after about 6 days. This can be compared to other dark/light ratios from the same

experiment shown in Figure 12.

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Figure 12. Cell concentrations with different dark:light cycles (Jacob-Lopes, 2009)

From this it can be seen that when a ratio of 2:22 is used a greater concentration

of algae can be attained than given constant light and a 4:20 ratio still gives an

acceptable concentration curve. Any larger dark aspect and the concentration begins to

diminish quickly with the exception of 12:12, which could be because it is a more natural

and balanced ratio. Both of these experiments give similar information curves for two

different strains of photosynthetic algae therefore these results can give an indication of

how many strains will behave in similar situations.

The other important aspect of growing the algae is not just the biomass

generated but also the CO2 that is removed. Jacob-Lopes also studied how different

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dark/light cycles affected the CO2 absorption. This is for the A. microscopica strain;

however these results can be compared to other photosynthetic organisms because the

underlying process is the same. Figure 13 shows a histogram of the CO2 activity.

Figure 13. CO2 absorption as compared to a 24 hour period of light (Jacob-Lopes, 2009)

This data set is with respect to a 24 hour period of light. As it can be seen the

longer the light is applied the more CO2 will be absorbed. From this it seems that

constant light should be used, however, if the algae is exposed to light constantly it

greatly increases the effects of photoinhibition. This data set has been given as a ratio

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of hours but this same ratio can be applied to seconds. For example 6:18 can be 6 hours

of dark and 18 hours of light and 6 seconds of dark and 18 seconds of light.

2.5 Compensation Point

The compensation point is the light level at which net photosynthesis is equal to

the rate of respiration. “For light level greater than the compensation point, the alga

will make a net gain in photosynthesis over losses due to respiration” (Graham &

Wilcox, 2000, p. 564). The compensation point is shown in Figure 14 at the intersection

point of the gross photosynthetic rate, Pm, and the zero net-O2 exchange line. Figure 14

shows these curves for Chlamydomonas reinhardtii, a green algae. The curves can be

defined by several different equations, two of which are:

3 � 34[1 − exp �:;34�] (4)

3 � 34 tanh(:;34) (5)

Where Pm is the maximum rate of gross photosynthesis, E is the light intensity,

and α is the slope of the initial increase in gross photosynthesis (Jassby & Platt, 1976).

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Figure 14. Light curve comparing light intensity to photosynthesis rates (Graham &

Wilcox, 2000)

2.6 Light Transmittance

When light passes through two different transparent media it will do two things;

part of it will reflect and the rest will refract. Reflection occurs when a beam of light hits

a surface and instead of moving through the surface moves back at the same angle. This

is illustrated in Figure 15.

Figure 15. Light being reflected at media interface

Compensation

point

Zero net-O2

exchange line

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Refraction occurs when the light beam passes through the interface but the

angle it leaves the first media is not the same as its entrance into the second, as shown

in Figure 16. These concepts can be described also by common applications, such as a

mirror for reflection and a lens for refraction. The amount of light that will pass reflect

or refract is determined by the indexes of refraction and the angle of incidence.

Figure 16. Light refracting (Weisstein, 2007)

The indices of refraction are based on how fast light travels through a substance.

The higher the index of refraction the slower light travels through that substance and

when light changes speed it bends back to the normal of the surface. The angle of

refraction can be found using Snell’s Law:

nC sin θC � nG sin θG

Where n1 = index of refraction of exiting media

n2 = index of refraction of entering media

θ1 = angle of incidence

θ2 = angle of refraction

(4)

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If the angle of incidence is changed the amount of light that is reflected or

refracted can be changed, even to the point of total internal reflection, wherein no light

is refracted.

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3 DESIGN APPROACH

3.1 Design Parameters and Constraints

There are several design considerations that must be addressed when designing

a bubble column reactor, the most significant being the availability of light throughout

the reactor. Because algae absorb light and also block light, light only penetrates

approximately 0.2 meters through fluid containing algae. Therefore, if the radius of an

externally lit bioreactor is larger than 0.2 meters there will be a dark zone in the center

as well as dimly lit areas near the center. This is unacceptable for sustainable algae

growth. The reactor can be larger if some kind of internal lighting is utilized, which is

the focus of this work.

The second design consideration is the introduction of CO2 to the system. For

this system in this work, CO2 was sparged into the system in the form of bubbles. To

increase the mass transfer properties based on the gas holdup value of the system,

small bubbles are necessary, because of the longer residence time and greater surface

area.

3.1.1 Fiber Optics

The choice of fiber optics for the light delivery system was based on the long

term goal of using a solar collector to provide natural light. A fiber optic illuminator was

used to simulate the properties of natural light, Figure 17, because the spectrum of the

halogen light bulb is similar to that of solar radiation (Davidson, 1999). The fiber optics

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are 3mm in diameter and are cut to necessary lengths. The ends of the fiber optics are

ground using a glass grinding wheel, Figure 18.

Figure 17. Fiber optic illuminator

Figure 18. Fiber optic polisher

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3.1.2 Gas Sparger

The sparging of gas into the system provides steady mixing of the solution and

introduces CO2. If the gas is sparged too quickly into the system the flow regime may

become heterogeneous, or turbulent, and the algae cells could potentially be harmed or

killed due to increased shear stresses. However, if the gas is sparged too slowly poor

mixing could result as well as suboptimal levels of nutrients (Miron A. S., Bubble-Column

and Airlift Photobioreactors for algal Culture, 2000). Homogeneous flow regime was

designed based on Figure 9 from section 2.3.

The main tube that makes up the column was 8” (0.20 m) in diameter to match

the dimensions of an airlift reactor previously built by the research group. Therefore to

remain in the homogeneous range the superficial gas velocity must be less than 2 in/s (5

cm/s). A sparger from Diffused Gas Technologies was purchased to serve as the

diffuser. This diffuser is a ceramic dome 7 ½” in diameter with a volumetric flow rate

between 0.5 and 2 CFM. This corresponds to gas velocity range of 0.0815 to 0.3262

in/s.

3.2 Algae Specimen

The algae strain used for this work was Scenedesmus dimorphus instead of

Chlorella vulgaris as it was unavailable. S. dimorphus is a fresh water green algae from

the regions of Australia and Saudi Arabia. Figure 19 shows S. dimorphus in solution

taken from the reactor during testing, including its relative size. The change in algae

specimen has a negligible effect on this work. Though the size of C. vulgaris is less than

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half of S. dimorphus, the chlorophyll a is the property being tracked and that is not

based on size of the organism. Also, the data used for the fluid properties was based on

the averages of many different green algae so the data is as valid for S. dimorphus as it is

for C. vulgaris. The scattering affect is also minimal as both species are microscopic and

the majority of scattering are due to macroscopic bubbles as will be shown in section

4.3.

Figure 19. Scenedesmus dimorphus in solution

3.3 Optical Simulations

The most important aspect of the design of the internally lit bubble column is the

lighting system. Software simulations were conducted to determine how light interacts

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with water containing live algae. By using software simulations to understand what is

happening in the reactor, time and money can be saved as many configurations can be

tested without the need to build each. Simulations, however, require a greater

knowledge of the processes and properties than building and testing a light scheme.

Therefore, it is important to determine the most accurate data sets to use for future

work. By comparing light intensities from the real world system to that of the

simulation model, it would be possible to assess the validity of the algae characteristics

used in the simulations.

3.3.1 Simulation Software

To determine the effectiveness of the light plates the optical simulation software

TracePro from Lambda Research was used (Lamdba Research). This program allows for

solid models to be imported from programs such as SolideEdge, and has very basic

modeling functions included in the package. Once a model is entered, the user then

defines the properties of the material and the light source and runs a simulation. The

results of the simulation are based on the scattering and diffraction of light for specific

input geometry, materials and fluid characteristics. The equations of the light models

are based on ray tracing done with the Monte Carlo Method. A Monte Carlo Method

involves random number sequences used to solve otherwise unsolvable sets of

differential equations.

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3.3.2 Simulation material properties

The most important components that need to be defined for the simulation are

the fiber optic cables, the light plates and the algae solution. TracePro includes an

acrylic material in the database so for the fiber optics and the light plates the material

was chosen as such. The algae solution was not included in the database and therefore

was created as part of the modeling work. Optical properties of light are broken into

two categories: apparent optical properties (AOPs) and inherent optical properties

(IOPs). IOPs are dependent on the media whereas AOPs are dependent on the light field

within the media.

Because photosynthesis does not require a specific orientation, AOPs are of

much less significance for this work than IOPs. The most important IOPs are the

absorption coefficient and the volume scattering function (Bass, 2010). Therefore it is

possible to create a simulation version of algae suspension and model photosynthetic

active radiation (PAR) attenuation, Figure 20, if these two properties can be fully

described. For use in TracePro the absorption coefficients, a(λ), and scattering

coefficients, b(λ), are needed in units of mm-1.

Scattering due to bubbles could not be inputted directly into the simulations

because the movement of the bubbles could not be simulated. Only steady state

systems could be worked on. Also, at the time of the simulations the characteristics of

the bubbles were not known because the sparger had not been selected yet. The

scattering due to bubbles does play a role in the dispersion of the light throughout the

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reactor but not in the removal of light from the system. This means that the affect is

not as significant because scattered light is still useful light, as opposed to absorbed light

that is used. The design of the light guide can proceed at this point without bubble

scattering considered because the scattering is consistent regardless of the design.

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Figure 20. Light attenuation calculations process (Bass, 2010)

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The absorption and scattering coefficients are given in the Handbook of Optics

through a review of past work by Andrea Morel that categorizes the coefficients based

on wavelength. Both coefficient sets are given in chlorophyll a specific values. This

allows the values to be used to determine the coefficients for any value of chlorophyll a

concentration. The chlorophyll a specific absorption coefficient was determined by

averaging 14 different phytoplankton from three different studies, Table 4.

Table 4. Chlorophyll a specific absorption coefficients with respect to wavelength (Bass,

2010)

λ (nm) Ac (m2 mg

-1) Λ Ac λ Ac λ Ac

400 .0394 475 .0347 550 .0098 625 .0061

405 .0395 480 .0333 555 .0090 630 .0063

410 .0403 485 .0322 560 .0084 635 .0064

415 .0417 490 .0312 565 .0078 640 .0064

420 .0429 495 .0297 570 .0073 645 .0066

425 .0439 500 .0274 575 .0068 650 .0071

430 .0448 505 .0246 580 .0064 655 .0084

435 .0452 510 .0216 585 .0061 660 .0106

440 .0448 515 .0190 590 .0058 665 .0136

445 .0436 520 .0168 595 .0055 670 .0161

450 .0419 525 .0151 600 .0053 675 .0170

455 .0405 530 .0137 605 .0053 680 .0154

460 .0392 535 .0125 610 .0054 685 .0118

465 .0379 540 .0115 615 .0057 690 .0077

470 .0363 545 .0106 620 .0059 695 .0046

700 .0027

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To determine the scattering coefficient, b, use Equation 5 (Bass, 2010)

H(I) � HJ(I) + 0.30 ∗ �550.NI � ∗ O�.PG (5)

Where C is the chlorophyll a concentration of the algae solution in mg/m3 and bw(λ)

values can be found in Table 18.

Table 5. Scattering coefficients (bw) for water and sea water (Bass, 2010)

λ

(nm)

Pure Water

(10-4

m)

Pure Sea

Water

(10-4

m)

λ

(nm)

Pure Water

(10-4

m)

Pure Sea

Water

(10-4

m)

350 103.5 134.5 475 27.6 35.9

375 76.8 99.8 500 22.2 28.8

400 58.1 75.5 525 17.9 23.3

425 44.7 58.1 550 14.9 19.3

450 34.9 45.4 575 12.5 16.2

475 27.6 35.9 600 10.9 14.1

To find the scattering coefficients for the wavelength range available for the absorption

coefficient the values in Table 2 are plotted, Figure 21 and the equation of the curves

found and used to find bw(λ) for Equation 5.

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Figure 21. Scattering coefficient curves based on data from Table 5

Table 6. Scattering coefficient with respect to wavelength for pure sea water

λ (nm) bw (m-1

) λ bw λ bw λ bw

400 .0781 475 .0398 550 .0203 625 .0103

405 .0747 480 .0370 555 .0194 630 .0099

410 .0714 485 .0364 560 .0185 635 .0094

415 .0683 490 .0348 565 .0177 640 .0090

420 .0653 495 .0332 570 .0169 645 .0086

425 .0624 500 .0318 575 .0162 650 .0082

430 .0597 505 .0304 580 .0155 655 .0079

435 .0570 510 .0290 585 .0148 660 .0075

440 .0545 515 .0278 590 .0141 665 .0072

445 .0521 520 .0265 595 .0135 670 .0069

450 .0498 525 .0254 600 .0129 675 .0066

455 .0476 530 .0243 605 .0123 680 .0063

460 .0455 535 .0232 610 .0118 685 .0060

465 .0435 540 .0222 615 .0113 690 .0057

470 .0416 545 .0212 620 .0108 695 .0055

700 .0053

y = 2.186e-0.009x

R² = 0.9907

y = 2.8598e-0.009x

R² = 0.991

0

0.02

0.04

0.06

0.08

0.1

0.12

0.14

0.16

300 400 500 600

Sca

tte

rin

g C

oe

ffic

ien

t (m

^-1

)

Wavelength (nm)

Pure Water

Pure Sea Water

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Table 7. Scattering coefficients with respect to wavelength for pure water

λ (nm) bw (m-1

) λ bw λ bw λ bw

400 0.0597 475 0.0304 550 0.0155 625 0.0079

405 0.0571 480 0.0290 555 0.0148 630 0.0075

410 0.0546 485 0.0278 560 0.0142 635 0.0072

415 0.0522 490 0.0266 565 0.0135 640 0.0069

420 0.0499 495 0.0254 570 0.129 645 0.0066

425 0.0477 500 0.0243 575 0.0124 650 0.0063

430 0.0456 505 0.0232 580 0.0118 655 0.0060

435 0.0436 510 0.0222 585 0.0113 660 0.0058

440 0.0417 515 0.0212 590 0.0108 665 0.0055

445 0.0398 520 0.0203 595 0.0103 670 0.0053

450 0.0381 525 0.0194 600 0.0099 675 0.0050

455 0.0364 530 0.0185 605 0.0094 680 0.0048

460 0.0347 535 0.0177 610 0.0090 685 0.0046

465 0.0333 540 0.0169 615 0.0086 690 0.0044

470 0.0318 545 0.0162 620 0.0082 695 0.0042

700 0.0040

A simplified version of the calculations done by TracePro is the Beer-Lambert law shown

in Equation 6.

Q � Q�(�R(S)TU Where (I) � �(I) + H(I)

(6)

Comparing a single ray analysis to a multi-ray analysis is not useful, however, as

an understanding of how light attenuates through a medium is important. An example

of the calculations shows this very important phenomena. For example, assume an

initial intensity of 200 µEinsteins at a wavelength of 425 nm. A data table, Table 8, can

be created to show a variety of results based on the two most important criteria,

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distance from the light source and concentration of chlorophyll a. For this example

(I) � 0.00243 and C and l are varied for different situations.

Table 8. Two variable analysis of light intensity using Beer-Lambert's Law

Initial

intensity Distance from light source (mm)

200 10 20 30 40 50

Co

nce

ntr

ati

on

of

Ch

loro

ph

yll a

5 177.14 156.90 138.97 123.09 109.02

10 156.90 123.09 96.57 75.76 59.43

25 109.02 59.43 32.40 17.66 9.63

50 59.43 17.66 5.25 1.56 0.46

75 32.40 5.25 0.85 0.14 0.02

100 17.66 1.56 0.14 0.01 0.00

3.3.3 Light plate design and testing

The light plates used for this work were developed based on a patent for

backlighting LCD televisions (Ciupke & al, 1993). The design concept, Figure 22, takes

advantage of Snell’s law in that the angled faces only allow light interacting with it from

a certain angle to pass through. A key design variable that needed to be determined

through simulation was the angled faces are 35, 45, 55, and 60° based on the angle of

incidence. The common dimension among all the plates is the top edge at 2” and the

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length of the plate various with the change in angle. Also, “long” versions of the plates

were tested to see if this change made a significant difference.

Figure 22. Short (left) and long (right) versions of the light plate (units in inches)

The bioreactor was modeled in Solidedge and imported to TracePro as a .SAT

file. Once in TracePro a light source and an observation disk were added to the model.

The light source was made to mimic the illuminator used in this work. The light consists

of 350,000 rays ranging from 400 to 700 nm. The observation plate is a transparent

object that is included in the model to act as a light sensor. The size of the observation

plate can be changed to encompass the cross section of the column and can be moved

vertically through the column. The light incident upon the surface of the observation

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plate can be selected for analysis as this would represent the light intensity in the

column.

The acrylic light pipes act as a modified light pipe to transport the light but also

to “spread” the light around. The light plate design utilizes Snell’s law by allowing the

light to leave the plate only through the angled faces and then only when the light

interacts with the angled faces at a certain angle. The light rays will “bounce around”

inside the plate until encountering the angled faces as shown in Figure 23.

To be considered a successful design, a light plate must provide at least 20

µEinsteins m-2 s-1 at a distance of 550 mm from the top of the reactor with a chlorophyll

a concentration of 50 mg/L. The light plate that exceeds this specification were

constructed and tested in a real world system.

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Figure 23. Light paths through the unique plate design

3.3.4 Benefits of light plates

There are several benefits that come from using rigid light guides and, more

specifically, the light plate design discussed here as compared to individual fiber optic

cables scattered throughout the reactor. Scattered fibers would provide very

concentrated light intensities at many places throughout the reactor. These intensities,

however, reach levels well above the saturation point for photosynthetic algae for the

space around the end of the fibers. Intensities of this level can cause harm to the algae

through the phenomenon of photohibiton, which from Figure 14, means an end to

photosynthesis caused by damage to the photon receptors which will shut down algae

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growth. Powles reported that light in excess of 300 µEinsteins/(m2s) will shut down

photosynthesis for several hours (Powles, 1984). Hincapie measured light intensities

near the tip of fiber optics to be as high as 600 µEinsteins/(m2s) (Hincapie E. , 2010) and

even with light scattering this intensity is still far above the saturation point. Therefore,

simply installing fibers throughout the reactor is not an efficient method of distributing

light.

The light plate design would take this high light intensity and distribute it over a

great area. The distribution area at the end of 5 fibers is approximately 0.07 in2,

whereas the plates have a distribution area of up to 41 in2. Most of the light passing

through the plate, however, will pass through the angled faces, with a surface area of

3.3 in2 which is nearly 50 times the distributive surface of 5 individual fibers. However,

assuming the light was evenly distributed through the entire plate, the intensity per

square inch would be in the range of 36-73 µEinsteins/in2, depending on the intensity

entering the plate from the 5 fibers. These light levels throughout the reactor will be

much more tolerable to the algae than the burst of light energy delivered by the fibers.

The light plate also provides a path for the light to travel through, which is not

constantly absorbing it. Therefore, more light needed for photosynthesis, blue and red

wavelengths, can be delivered at more locations deeper inside the reactor. The light

distribution for any light guide can be observed with the use of photodiodes.

Photodiodes convert light energy to electrical energy which can be measured. By doing

this all of the light can be accounted for. This will be discussed more in Chapter 6.

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Another benefit is the rigid positioning of the plates that is not as easily achieved

with individual fibers. If not securely fashioned, individual fiber optics would tend to

move based on hydrological forces and this would result in an unpredictable light

distribution and could possibly cause the fibers to pull away from the illuminator. To

secure the fibers to prevent any movement and to ensure that light patterns could be

maintained, there would have to be a substantial structure in place. This structure

would simply take up volume in the reactor as well as introduce complex issues of

shading. The setup utilized for this work only required 1.3L and there were no issues

with shading as all of the support structure were positioned on the outside edge of the

reactor.

3.4 Reactor Design

Based on the simulation results, as discussed below, the layout of the reactor

consists of two layers of four light plates. The light plates follow the 35° design and are

approximately 15” long, as shown in Figure 24. The plates were modeled in Mastercam

X and machined using a CNC milling machine to ensure the accuracy needed for proper

light distribution. The top performing plate in the simulation measured more than 17”

in total length however the plate length had to be reduced to 15” to fit within the

maximum travel on the CNC mill in our machine shop.

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Figure 24. 35° light plate to be installed in BCR

The light plates are held in place using polyester disks, Figures 25 and 26, which

hang on the inside of the column. The bottom disk is designed to allow enough support

to hold the light plate while also allowing for minimum influence on fluid flow.

Figure 25. Bottom plate holder

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Figure 26. Top plate holder

The bottom disk was hung by securing four support rods, Figure 27, to the disk at

the countersink locations. The top circular plate rest on a shoulder machined onto the

support rod. The light plates on the bottom were fed fiber optics via a transport tube

that was dropped down directly on top of them. The support rods were screwed to the

blind flange at the top of the reactor to secure and square them. The transport tubes

pass through the blind flange and are supported by the stoppers located at the top of

the tube.

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Figure 27. Partially constructed components of BCR

The plates are held using a slot and bolt mechanism shown in. This design allows

for easy assembly and removal for cleaning and maintenance purposes. Figure 28

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shows the mechanism for the lower disk and Figure 29 shows the mechanism for the

upper disk. The only difference is the addition of the added step to further secure the

lower plates because the only points of contact for them are the two tabs and a small

slot on the lower disk.

Figure 28. Slot mechanism for lower disk

Figure 29. Slot mechanism for upper disk

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The diffuser chosen for this reactor is the EW-70025-20 model from Diffused Gas

Technologies, Figure 30. This model is a fine bubble air stone that can maintain the

required superficial gas velocity for a homogeneous bubble flow regime. The diffuser

connects through the bottom of the reactor, Figure 31, and provides bubbling to the

entire cross-section of the column.

Figure 30. Gas delivery system

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Figure 31. Gas delivery system (looking from under the reactor)

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Figure 32. Internal light components prior to assembly with column

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Figure 33. Lighting components installed in column

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3.5 Illuminator Cooling

To attempt to minimize the damage to the fiber optics done by the heat from the

illuminator a cooling system was developed utilizing aluminum fins and quartz. Two pieces of

quartz were embedded in the aluminum with an air gap, Figure 34. Also, the upper section of

the illuminator housing was pulled back to allow for forced convection cooling from a fan. The

heat buildup was substantially lower with this added cooling capabilities however, the heat is

still an issue.

Figure 34: Cooling system for illuminator

3.6 Reflective Coating

Through initial testing before inoculation it was observed that light levels

according to the sensor were considerably low compared to expected values from

optical simulations, in a range of 6-8 µEinsteins at the 40 cm mark. It was observed that

light was exiting the light plate through the sides and back of the plates and this is

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believed to be contributing to the low light levels. It was also observed that light was

leaving the column at a much greater rate than was expected from the optical

simulations. In attempts to understand the low light intensity values the reactor was

wrapped in aluminum foil to ensure light remained in the column, Figure 35.

Additionally, the light plates on the top level were also covered in aluminum foil to

direct the light in a way that recreates the simulation conditions, Figure 36.

Figure 35. BCR covered in aluminum foil

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By wrapping the column in aluminum foil, light is much less likely to escape out

of the column and is instead redirected back into the column. The result of this change

is that the average light intensity measured in the reactor at the 40 cm mark is in the

range of 11 to 14 µEisnsteins. The wrapping also acts as a barrier from the growth lights

over the Scenedesmus tanks in the same room so the light measured was light coming

through the light plates.

Figure 36. Light plates covered in aluminum foil except over the angled faces

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The use of aluminum foil on the light plates further increased the light intensity

levels to approximately 20 µmols. The concern with this however is that the aluminum

foil could not be used when the column was inoculated. This situation is discussed

further in Chapter 6.

3.7 Test Plan

To validate the simulation results, the light intensity was measured in a working

reactor along with other vital fluid characteristics. The reactor, 30 L total volume, was

inoculated with six liters of algae solution from an incubator with a chlorophyll a

concentration of 46.4 and turbidity of 16.3 on 8/15/2011, the remainder of the volume

was DI water. 15mL of growth media, the recipe is found in Appendix A, was also added

at this time. The reactor was bubbled with 1% CO2 at a pressure of approximately 1.5

psi with the lights on continuously. Fluid measurements were begun on 8/16/2011 and

continued through 8/22/2011. Testing involved two essential components, the fluid

properties and light intensity.

3.7.1 Analytical Procedures for fluid properties

To determine the characteristics of the algae solution several other tests were

performed on the solution: turbidity, pH, and nitrate and phosphate levels. Samples

were drawn from the top of the reactor via a turkey baster and emptied in a beaker.

Turbidity is the measure of water clarity based on how the material suspended in the

solution affects light passage (Lenntech, 2011). Turbidity can be caused by particulates

such as nitrates and phosphates, bubbles, and dead and living algae cells. A turbidity

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measurement can be used in conjunction with the chlorophyll a measurement to better

understand what the cells are doing as a chlorophyll a measurement does not give

details on the amount of cells. The turbidity of the system was measured using the

Hach 2100P turbidimeter (Hach, 2011)

The limiting reagents in algae growth are usually nitrate and phosphate.

Therefore these compounds must be monitored periodically. To measure these

compounds the Hach DR/890 colorimeter was used. The algae mass was filtered out of

the solution and the remainder was split into two vials. In one vial a compound for

nitrate measurement was added and a compound for phosphate was added to the

other. The vials are then put into the colorimeter for measurement. The colorimeter

uses light to detect mineral concentrations by emitting wavelengths of 420, 520, 560,

and 610 nm through the fluid and detected what light passes through (Hach, 2011). The

nitrate compound contains cadmium which “reduces nitrates present in the sample to

nitrite. The nitrite ion reacts in an acidic medium with sulfanilic acid to form an

intermediate diazonium salt which couples to gentisic acid to form an amber-colored

product (Hach Company, 2005).” and the phosphate compound contains

orthophosphate which “reacts with molybdate in an acid medium to produce a

Phosphomolybdate complex. Ascorbic acid then reduces the complex, giving an intense

molybdenum blue color (Hach Company, 2005).” These colors allow the colorimeter to

measure the concentration of ions in solution.

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The pH was measured using an Accumet pH meter. Because CO2 was bubbled

through the reactor the pH will tend to be more acidic. The algae will tend to keep the

solution basic through their growth processes, however until this point is reached the

pH was stabilized using 1 m NaOH added when the pH falls below 6.8. The remaining

sample after testing was sterilized with bleach and disposed of to prevent

contamination of the reactor.

3.7.2 Light Measurements

To measure the light penetration and intensity in the reactor, a rig was

developed to hold the LI-192 light sensor in the center of the system, Figure 37. The

sensor must be oriented upward, Figure 38, as preliminary tests showed that the sensor

does not measure light coming from behind the sensor.

Figure 37. Light sensor fixture including depth rod

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Figure 38. Sensor orientation inside reactor for accurate light measurements

The light sensor was moved through the reactor via the depth rod. The depth

rod was marked every 2 cm from 24 to 40 cm with respect to the top of the reactor,

Figure 39. When comparing this depth to the depth in the simulation 5 cm must be

added because of the way the simulation was constructed. Therefore, 40 cm

corresponds to 45 cm in simulation. Knowing the intensity at various depths, the light

penetration can be studied as factor of depth and distance from the light source. The

primary light source in the reactor is the surfaces of the light plates. So when the sensor

is lowered it is steadily moving away from the light source. Figure 40 shows distances in

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the upper region of the reactor so to understand where in the reactor the sensor is

when using the depth rod.

Figure 39. Depth rod with markings shown

Figure 40. Distances between various components of the reactor to understand

placement of sensor using depth rod marking. The sensor in this case is at a depth of 24

cm. Illustration is not scale.

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Each set of measurements included the average light intensity taken over ten

seconds with and without bubbling. The only values that could have been compared to

the simulation results are the intensities without bubbling. The simulations were done

without factoring in scattering due to bubbles. The intensities taken can give an

understanding of the impact of bubbling which can then be implemented into future

simulation models.

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4 RESULTS AND DISCUSSION

To determine the effectiveness of the light plate design, simulations were

conducted to study light penetration and attenuation through algae media. Then, the

light plate that performed the best according to the criteria was constructed and

implemented into the system. The system was tested to validate the input parameters

(such as the characteristics of the media) and the output of the simulation.

4.1 Simulation Results

Eight variations of the light plate were tested using the optical simulation

software TracePro. These variations include “short” and “long” versions of plates with

35°, 45°, 55° and 60° angles of incidence as discussed in Chapter 3. The results are

displayed in Figures 40 and 41. While only chlorophyll a concentrations of 50 and 100

mg/m3 are shown on the graph for visibility, concentration of 10 mg/mm3 was also

evaluated. The chlorophyll a concentration can be used as a relationship to the actual

biomass concentration as it describes the production levels of the algae. This is also a

vital measurement as the absorption of light by the chlorophyll a pigment is the factor

with the greatest influence on light penetration and propagation as shown in Tables 4-7.

These results show how well distributed the light from the light plates can be.

For these simulations the minimum light at the 550 mm mark must be greater than 20

µEinsteins/m2s using a chlorophyll a concentration to be a qualifying design. The only

light plate that achieved this was the long version of the 35° plate. This was not an

expected outcome as according to Snell’s law the critical angle for reflection is 60°. This

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is most likely due to the fact the light remained in the plate longer in the 35° than the

60°. Another benefit of the 35° long plate is that it produced the highest light intensity

for 100 mg/mm3 of chlorophyll a at 550 mm. While the difference is relatively small any

additional light distribution is desirable to decrease the amount of energy input needed.

The light distribution for the 35° “long” is also much more conducive to growing

algae as it provides dark areas for the algae. The other plates do not allow sufficient

opportunities to pass through the dark areas. Figures 41-42 shows the light distribution

for the 35° long plate system. This is an important design aspect because many times in

the light distribution there are very bright spots and mostly darkness and this warps the

average. Figure 43 shows a fairly even distribution of light with a few bright spots but

most of the spots are in the 1000-4000 lux range. For these simulations the unit lux was

chosen over W/m2 because it can be more easily compared to µEinsteins on a PAR basis

(Licor, 2010). The other important design criterion is the wavelength of light that

penetrates. Due to the nature of photosynthesis, ranges of blue and red wavelengths

are absorbed and therefore do not penetrate as deep as green light. Figure 43 also

shows the distribution of light based on the wavelength color. The most important

thing to look for in this type of plot is to see if there are blues and reds in that cross

section. The design of the light plate allows for these wavelengths that are absorbed

quickly to be constantly replenished.

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Figure 41. Results of light simulations for two different chlorophyll a concentrations throughout the reactor. (a) 35 plate, (b) 35 long

plate, (c) 45 plate, (d) 45 long plate

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Figure 42. Results of light simulations for two different chlorophyll a concentrations throughout the reactor. (a) 55 plate, (b) 55 long

plate, (c) 60 plate, (d) 60 long plate

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Figure 43. Light intensity distribution (Left) and color map (right) of the 35° long plate at a depth of 425 mm

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4.2 Reactor results

To validate the simulation work, light tests were performed on the functioning

reactor. Light measurements were taken at 10 a.m., 1 p.m., and 4p.m. along with

measurements of pH, turbidity, and chlorophyll a concentration. Because algae

properties can change relatively quickly, measurements taken several times a day and

averaged together is a more accurate way of examining the growth of the algae rather

than taking several measurements at one point in the day.

The results of the measurements are shown in Tables 9-15. The tables show

light intensities measured at steady state (no bubbling) and with homogeneous flow

(with bubbling). It is clear that the scattering due to bubbles does limit the light

penetration, but not at a severe enough rate to disqualify the light plate design. The

diffusive nature of the bubbles may serve to better distribute light to areas that

otherwise may not have received any. The reactor was inoculated on 8/15/2011 and

the first measurements were taken on 8/16/2011. It was decided on 8/17/2011 to take

light measurements with and without bubbling to understand the impact of bubbling.

The intensity was measured in each state without moving the sensor and by just

opening or closing the air regulator to ensure that the sensor location was consistent.

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Table 9. Results from 8/16/2011

Light intensity (µmol m-2 s-1) measured from the top of reactor

Time chlorophyll

a turbidity pH bubbling 40 38 36 34 32 30 28 26 24

µg/L yes/no cm cm cm cm cm cm cm cm cm

10:00 11.6 4.85 5.75 yes 2.61 3.13 3.11 3.46 3.16 3.63 3.72 4.01 4.04

no

1:00 11 5.42 6.8 yes

no 2.41 2.38 2.37 3.9 4.2 4.6 4.92 5.26 5.15

4:00 11.8 4.93 7.041 yes

no 8.78 8.74 9.14 9.45 9.96 9.55 9.78 9.16 9.27

Table 10. Results from 8/17/2011

Light intensity (µmol m-2 s-1) measured from the top of reactor

Time chlorophyll

a turbidity pH bubbling 40 38 36 34 32 30 28 26 24

µg/L yes/no cm cm cm cm cm cm cm cm cm

10:10 13.6 5.13 6.98 yes 5.6 5.75 6.64 7.48 7.59 8.3 8.23 7.51 7.93

no 7.02 7.07 7.81 8.6 8.61 9.14 8.74 8.03 8.13

12:50 13.1 4.97 7.066 yes 5.56 5.79 6.05 6.5 6.82 7.05 6.82 7.02 6.28

no 6.97 7.13 7.29 7.47 7.62 7.67 7.13 6.98 5.9

4:00 14.2 4.77 7.024 yes 6.51 6.67 6.79 7.76 8.38 7.31 8.65 8.75 9.3

no 8.23 8.29 8.25 9.12 9.72 9.93 9.6 10.15 8.97

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Table 11. Results from 8/18/2011

Light intensity (µmol m-2 s-1) measured from the top of reactor

Time chlorophyll

a turbidity pH bubbling 40 38 36 34 32 30 28 26 24

µg/L yes/no cm cm cm cm cm cm cm cm cm

10:30 15.1 4.71 7.004 yes 6.69 6.77 7.01 7.88 8.36 9.57 8.36 9.26 10.17

no 8.09 8.11 8.15 9.11 9.48 10.73 9.2 9.79 10.49

1:00 15.4 4.99 7.06 y 7.48 7.47 7.57 8.64 8.06 8.1 8.99 10.22 10.38

no 9.03 8.96 8.94 10.11 9.21 9.16 9.98 10.86 10.79

4:00 14.6 4.44 6.988 y 9.46 9.53 7.81 8.15 8.56 8.28 8.54 6.88 6.59

n 9.98 10.18 8.62 9.21 9.63 9.52 9.96 8.33 7.92

Table 12. Results from 8/19/2011

Light intensity (µmol m-2 s-1) measured from the top of reactor

Time chlorophyll

a turbidity pH bubbling 40 38 36 34 32 30 28 26 24

µg/L yes/no cm cm cm cm cm cm cm cm cm

10:05 16.6 5.48 6.971 yes 6.03 6.37 6.69 7.2 7.68 8.87 7.8 7.05 7.17

no 7.35 7.6 7.94 8.47 8.69 10.29 8.64 7.65 7.66

1:45 16.6 5.56 7.025 yes 6.74 6.64 6.77 7.71 7.64 7.3 6.94 7.08 6.92

no 7.91 7.92 7.88 8.97 8.6 8.05 7.6 7.79 7.18

4:00 17.3 5.71 6.997 yes 6.55 7.28 8.53 8.86 8.04 7.82 8.35 6.95 6.91

no 7.6 8.44 9.72 9.81 8.86 8.43 8.85 7.39 7.12

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Table 13. Results from 8/20/2011

Light intensity (µmol m-2 s-1) measured from the top of reactor

Time chlorophyll

a turbidity pH bubbling 40 38 36 34 32 30 28 26 24

µg/L yes/no cm cm cm cm cm cm cm cm cm

10:20 18.5 6.39 6.97 yes 5.72 6.12 6.95 7.19 6.98 7.08 7.79 7.7 6.37

no 6.66 7.19 7.99 8.18 7.87 7.83 8.17 8.09 6.45

1:05 20.1 7.27

7.024 yes 6.02 6.35 7.65 7.98 8.49 8.46 8.43 7.68 7.72

no 7.14 7.36 8.56 8.83 8.99 8.82 8.57 7.76 6.98

4:00 19.9 6.3 7.01 yes 5.47 5.92 6.5 6.97 8.21 9.18 7.76 7.87 8.12

no 6.47 6.94 7.47 7.95 9.08 9.92 8.3 8.46 8.55

Table 14. Results from 8/21/2011

Light intensity (µmol m-2 s-1) measured from the top of reactor

Time chlorophyll

a turbidity pH bubbling 40 38 36 34 32 30 28 26 24

µg/L yes/no cm cm cm cm cm cm cm cm cm

10:10 21.3 6.91 7.005 yes 5.44 5.81 6.89 7.15 7.77 7.85 7.41 7.26 7.02

no 6.42 6.83 7.88 8.01 8.47 8.5 7.76 7.39 7.28

1:05 21.3 6.78 7.016 yes 5.57 5.43 5.95 5.99 6.62 7.14 7.5 7.76 7.45

no 6.18 6.34 6.81 6.82 7.32 7.71 7.86 7.59 7.17

4:00 23.8 7.3 7.017 yes 5.24 5.6 5.44 7.17 7.38 7.71 7.24 7.94 7.32

no 6.2 6.5 6.31 8.1 8.15 8.37 7.67 8.27 7.35

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Table 15. Results from 8/22/2011

Light intensity (µmol m-2 s-1) measured from the top of reactor

Time chlorophyll

a turbidity pH bubbling 40 38 36 34 32 30 28 26 24

µg/L yes/no cm cm cm cm cm cm cm cm cm

10:10 22.8 8.54 7.008 yes 4.5 4.89 4.91 5.53 6.42 7.26 7.01 7.19 6.53

no 5.37 5.83 5.67 6.37 7.15 7.94 7.68 7.65 6.89

12:55 22 8.42 7.074 yes 4.59 5.03 5.98 6.23 6.6 7.17 6.23 7.49 6.9

no 5.33 5.93 6.9 7.11 7.4 7.84 6.9 7.92 7.22

4:10 20.8 7.81 7.175

yes 6.75 7.39 7.76 7.44 7.63 7.86 6.24 5.25 4.75

no 6.88 7.64 8.33 8.07 8.42 7.86 7.16 6.17 5.62

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There are several reasons that might explain why the actual test results did not

closely match the simulation results. The first possible source of discrepancy is that the

light sensor has not been calibrated in five years. Licor suggests that their sensors be

calibrated every two years. The sensor was compared to the LI-190 light sensor, Figure

44, in air and measured fairly close values as was seen in Figures 45 and 46; however,

this correlation cannot be applied to the sensor when it is underwater.

Figure 44. LI-192 underwater light sensor (Left) and LI-190 light sensor (Right)

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Figure 45. Comparison of calibrated and uncalibrated light sensor at a distance of 1.5"

from light source in air

Figure 46. Comparison of calibrated and uncalibrated light sensor at a distance of 4"

from light source in air

y = 1.8578e1.3711x

R² = 0.995

y = 1.7594e1.3595x

R² = 0.9932

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

0 1 2 3 4 5 6 7

Lig

ht

inte

nsi

ty (

mic

ro m

ols

)

illuminator marks

uncalibrated

calibrated

y = 0.3718e1.361x

R² = 0.9957

y = 0.3448e1.3675x

R² = 0.9962

0

200

400

600

800

1000

1200

1400

1600

0 1 2 3 4 5 6 7

Lig

ht

inte

nsi

ty (

mic

ro m

ols

)

Marks on illuminator

uncalibrated

calibrated

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The second source of discrepancy is the light patterns in the light plates. In

simulation, most light exited light plates through the angled faces with only negligible

amounts leaving the sides and back. However, in the real world system a great deal of

light leaves the sides and back of the light plates. This is not necessarily a detrimental

aspect of the design; it simply makes a comparison between the simulation model and

the real world system more difficult. Before the reactor was inoculated, the light plates

were wrapped in aluminum foil and the light intensities were measured. With the

aluminum foil acting as a reflector, the light intensity increased by as much as 100%.

Therefore, a reflective coating could bring the light intensity in the target area closer to

the goal intensity.

A third possible source of discrepancy is how the fiber optics connect to the

illuminator. The way the fiber optics pass through the coupler presents the possibility

that some fiber optics are shaded by the others. This means that not all the light is

reaching the reactor whereas in the simulation all of the fibers were given clear line of

sight of the bulb surface. Also, the temperature of the light bulb exceeds the melting

point of the acrylic fiber optics. This results in melting at the tip of the fiber optic, Figure

47. Melted fiber optic tips drastically decrease the amount of light that can be

transmitted through them. This melting occurred after additional heat sinks and cooling

were implemented. These issues are discussed in greater detail in Chapter 6.

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Figure 47. Melted fiber optic cable after prolonged use with the illuminator

A fourth issue could be the data that was used to construct the simulation media

was not appropriate for the strain of algae used or the chlorophyll a concentrations that

were handled in simulation. The absorption coefficient for chlorophyll a could be higher

than was used in the simulations, like the data shown in Figure 48; however I was

unable to run simulations with this new data. If the absorption coefficients are too low

then the penetration will appear higher than it is in the real world. It could also be that

the absorption of light by chlorophyll b played had a higher impact than anticipated.

Both the original and new set of data are just starting points and are not useful in final

analysis and therefore a thorough investigation of the absorption coefficients for a

single strain of algae should be done. This testing will be detailed in Chapter 6. Even

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with the higher absorption coefficient, it is possible to reach the 20 µEinsteins m-2 s-1

criteria established in section 3.5.3 by applying some of the recommendation that will

be presented in Chapter 6.

Figure 48. Absorption coefficients for water containing phytoplankton for chlorophyll a

concentration from 0.2 and 18.4 mg m-3 (Prieur & Sathyendranath, 1981)

Even though the light intensity was lower than expected, the algae grew at a

constant rate. The pH was maintained at approximately 7 with 1% CO2, the

temperature was regulated via the room air handlers, and the nutrient levels were

Wavelenth (nm)

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maintained. This means that the only limiting factor could have been the lighting. The

intensity values were above the compensation point for green algae, which is between 2

and 6 µEinsteins m-2 s-1 (Graham & Wilcox, 2000). Figure 49 shows the changes in

chlorophyll a and turbidity over the time of the experiment.

Figure 49. Chlorophyll a and turbidity changes over course of experiment. Inoculation

occurred at 7 p.m. on Day 0 with the first measurements taken on Day 1.

The chlorophyll a concentration increased due to the changes in the lighting. At

lower light intensities the algae will create more chlorophyll a pigment, this can be seen

in the first three days. Then as the number of algae cells increase the amount of

pigment continues to increase. The turbidity however, is a much more accurate

representation of the cell activitiy than just chlorophyll a. The first three days showed a

y = 1.881x + 9.6857

R² = 0.9729

y = -0.1767x + 5.2656

R² = 0.9547

y = 0.8363x + 2.2727

R² = 0.9604

0

5

10

15

20

25

0 1 2 3 4 5 6 7 8

Ch

loro

ph

yll

a c

on

cen

tra

tio

ns

(µg

/L)

an

d

turb

idit

y m

ea

sure

me

nts

Testing day

average…

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slight decrease in the turbidity of the water which would imply a slight decrease in the

algae concentration. This is expected as the algae adjusted to the new environment.

After the initial adjustment time the algae began to grow at a steady constant rate. The

reator had been run indefinitely it is possible that the chlorophyll a concentration would

have reached and surpassed the 50 µg/L used in the simulations. This proves that the

lighting system does provide lighting in both an intensity and distribution that promotes

algae growth.

The chlorophyll a and turbidity measurements may not be the most accurate

representation of the reactor. A test was performed on different growth tanks also

growing S. dimorphus to determing if taking samples from the top of a reactor resulted

in different values than samples taken from the bottom of a mixed reactor. The results

of these tests are shown in Table 16.

Table 16. Difference in measurements when samples are taken from the top and

bottom of the reactor

Location of sample

Sample #

type of

measurement Bottom Top

1 chlorophyll a 93.3 µg/L 59.1

turbidity 113 38.7

2 chlorophyll a 37 15.1

turbidity 39 20.9

3 chlorophyll a 110.1 52.3

turbidity 250 43.5

4 chlorophyll a 53.5 13.2

turbidity 138 22.5

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While no direct comparison can be made from Table 16, it can be seen that there

is a large difference in values depending on where the sample is taken from even in a

mixed reactor. Knowing that the change is considerable, this makes the growth values

taken from the bubble column reactor much more comparable to other reactors.

4.3 Scattering due to bubbles

An important phenomenon in the study of photobioreactors is the effect of

bubbling on light penetration. To further understand this light intensity was measured

at each depth with and without bubbling. The percent difference of these conditions

changes linearly with respect to depth based on Figure 50. The bubbles do reduce the

penetration depth of the light; however it is important to compare with the steady state

conditions. The same flux that is present in the steady state is within the system with

bubbling, the light is simply directed elsewhere. The main difference in this system from

externally lit systems with regard to bubble scattering is that the light in this reactor is

sourced on the inside, so the light remains in the reactor. With an external lighting

source, light is reflected back out of the reactor and therefore is wasted.

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Figure 50. Difference in light penetration with respect to distance based on bubbling.

The points are the day and time of the measurement, i.e. 1-10 was taken at 10:00 on

the day one of testing.

Because the plot values change linearly during the time that algae was growing

nonlinearly, it can be seen that the bubbles are the major source of light scattering, not

the algae cells. There was a small amount of variability over the week of testing that

could be attributed to scattering due to algae. This short test was not enough to prove

the exact impact of the bubbles, but was important to identify a future area of study.

4.4 Circulation

Circulation was evaluated using a dye test. Dye was injected into the system and

was videotaped while mixing. The dye test was done with all components installed

including the light sensor. However, light was not applied to the system for this test.

The dye is a red fluorescent tablet from Bright Dyes that was dropped from the top of

y = 0.6647x - 10.629

R² = 0.9666

-15

-10

-5

0

5

10

15

20

25

30

23 25 27 29 31 33 35 37 39 41

Pe

rce

nt

dif

fere

nce

Distance from top of reactor (cm)

17-10 17-1 17-4 18-10 18-1

19-10 19-1 19-4 20-10 20-1

20-4 21-10 21-1 21-4 22-10

22-1 22-4 average

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the reactor and allowed to sink (Bright Dyes, 2011). The video shows that mixing

beginning along the path of the dye tablet but then moves to the center of the reactor

and radiates outward. The entire reactor is mixed in approximately 46 seconds as

shown in Figure 51.

The circulation of the fluid was important for two main reasons. The first was

the exchange of matter from between the algae and the water. If the cells were not

moved they would not have access to the nutrients in the system as well as the CO2.

The second reason was to move the algae in and out of lighted areas. The reactor was

run with the illuminator on constantly. Photosynthesis however requires time when the

light intensity is below the compensation point that was discussed in section 2.5. The

moving fluid carried the cells in and out of contact with the light. During the test, eddy

currents could be seen that created a twisting motion through the solution. These eddy

currents appeared to be random in their appearance and direction.

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Pre-test condition

Dye tablet added to

reactor

Time: 0 seconds

Bubble reach the

top of reactor

Time: 10 seconds

Bottom half of

reactor is mixed

Time: 22 seconds

Reactor is fully mixed

Time: 46 seconds

Figure 51. Progression of mixing via a dye test

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5 CONCLUSIONS

The objective of this work was to develop a functioning bubble column

bioreactor with an internal lighting system in which algae could grow. To achieve these

objectives, optical simulations were conducted and a reactor was built based on the

results of the simulations. The design specifications for the simulations was a minimum

of 20 μEinsteins m-2 s-1 at a distance of 55 cm with a chlorophyll a concentration of 50

mg/L to create an environment in which algae can grow.

5.1 Working System

The proposed flow regime for the reactor was homogeneous based on the

literature review and Figure 9. To achieve this, an air pressure of 1.5 psi was used with a

mixture of shop air and 1% CO2. The result was a homogeneous flow with small bubble

size. The homogeneous flow was also evident from the dye test discussed in section

4.4. The small bubbles increased the gas holdup of the system and the mixing

capabilities with clearly evident eddy currents that appeared random in occurrence,

position, and direction of flow. The circulation resulting from this fluid flow is consistent

with reactors found in literature and is suitable for sustaining algae growth. There was

also only negligible algae adherence to the component surfaces.

The algae growth, given in terms of chlorophyll a and more importantly turbidity,

Figure 49, shows that when the testing ended the algae was still in a growth phase. The

numbers appear low when compared to the airlift reactor or to the other incubators;

however, as was discussed in section 4.2 this could be due to the location of the sample.

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5.3 Light Plate Design

The specification for the light plate design during simulation was that an array of

4 plates must provide at least 20 μEinsteins m-2 s-1 at a distance of 55 cm from the top of

the reactor filled with a solution with a chlorophyll a concentration of 50mg/L. The only

light plate arrangement that met this criterion was the 35° “long”, shown to give an

intensity of approximately 25 μEinsteins m-2 s-1 in Figure 41 in section 4.1. The light

intensity curves followed an expected pattern that decreased quickly and leveled off the

deeper the sensor was located. In the simulation the principal method of light

dispersion from the light plate was through the angled faces toward the middle of the

reactor, Figure 23 in section 3.3.3. This was the expected light path based purely on

Snell’s law and a simplistic view of the interaction between light, acrylic, and the growth

media.

The results from the real world reactor testing were given in Tables 9-15 in

section 4.2. These values were lower than expected from the simulation results with

the highest intensities being around 10 µEinsteins m-2 s-1 at a chlorophyll a

concentration of 15.1 mg/L. I was unable to test a chlorophyll a concentration of 50

mg/L as the solution never got to this level as the highest chlorophyll a concentration

was attained 23.8 mg/L. I was therefore, unable to meet the objective of replicating the

simulation results in the real world system. The light sensor used in this experiment cast

some doubt on the measurements taken due to the type of sensor and the number of

years since it was calibrated. Also, the simulation results were very rigid, as no light left

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the light plates through the sides; all of it exited through the angled faces. In the real

world system, light did leave through the sides and the back. Therefore, there was not

necessarily less light in the system, it simply did not match the simulation model as the

light was more diffuse and filled the reactor rather than just lighting the middle. There

are other possible reasons for this behavior and these were addressed in section 4.2.

Nevertheless, the light intensity measured in the reactor never fell below the

compensation point for green algae as discussed in section 2.5

As was described in Chapter 4, light was controlled to be the limiting reagent.

However, because the algae grew under the presented conditions, the lighting system

performed successfully regardless of the inability to replicate the simulation results.

The reactor met all of the requirements for sustaining algae growth by means of proper

mixing and CO2 availability. Also, knowledge and experience gained from this work

regarding internal lighting systems will add important information to the scarce amount

of data available in this field.

This project achieved all of the stated objectives and ISEE now has a working

internally lit bubble column reactor to compliment the internally lit airlift reactor for

future studies.

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6 RECOMMENDATION AND FUTURE WORK

6.1 Recommendations

The following are modifications or additions to the reactor that should be

considered for any future work. Following these recommendations the reactor could

function more in line with the simulation results. The recommendations include

changes to the lighting system and component materials that have been identified

during this work.

6.1.1 Lighting system

The first and most important recommendation for future work on this project

would be the purchase of an LI-193 Spherical underwater sensor from Licor, Figure 52.

This is the only sensor of its kind available on the market except for very large light

sensors that are used in ocean research. The LI-193 allows for a better collection of light

in algae solutions as it can sense light from all directions (Underwater PAR

Measurement, 2011). The LI-192 can detect light incident from the hemisphere above

the sensor surface, however this is a severe limitation in setup as it has to be located in

the exact same position and orientation each time. Any disturbance will change the

value read. This limitation has also been noted by a group of German optical scientists

when they compared various light sensors (Hartig and Heinrich, 1999). By using the LI-

193, the orientation of the sensor is negligible especially when exposed to diffuse light.

The LI-193 could be implemented in the same way in the reactor that the LI-192, with

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the bulb up as this has been shown in this work to be a stable method of holding the

sensor.

Figure 52. LI-193 Spherical Light Sensor (Underwater PAR Measurement, 2011)

Also, the illuminator needs to be modified or a better light source needs to be

found The light opening on the illuminator is too small to allow all the fiber optics the

appropriate lighting area. The heat from the bulb also degrades the fiber optics and

prohibits light transmission. An improvement that still uses the full visible range of the

light spectrum, to simulate natural sun light, is a metal halide. The metal halide has

several very important advantages to the halogen. A 150W halogen has an efficiency of

18-20 lumens/Watt, whereas a 150W metal halide bulb has an efficiency of 65-115

lumens/Watt. The metal halide also illuminates with a color temperature more light the

sun (Venture Lighting International, 2011) which Hincapie discussed as an important

parameter on algae growth (Hincapie E. , 2010). A halide bulb also operates cooler than

a halogen bulb (Edmund Optics, 2011). The cost of implementing such a system would

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be similar to the halogen illuminator presently in service; however, the bulbs are more

expensive. Figure 53 shows a fiber optic halide illuminator that is recommended for

future work with this reactor. It is a 150W illuminator that comes with an on/off

remote.

Figure 53. Metal halide fiber optic illuminator (Wiedamark, 2011)

A more detailed analysis of the potential of the light plates needs to be

performed as well, especially comparing to individual fibers. This needs to be done to

account for all the light leaving the plates and in what distribution. A method of

determining the light distribution for the optics and light plate comparison would be to

construct a box filled with photodiodes on the inside walls. Photodiodes convert light

energy to electric energy in the form of current or voltage which could be read with a

simply multimeter, or set up the entire array to be measured using a visual

representation on a computer screen. This method would be much more efficient than

using the simulation software TracePro again.

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6.1.2 Reactor components

Another modification to this system should also be to find a method of applying

a reflective surface to the light plates. As stated in Chapter 3, light was leaving surfaces

unexpectedly when compared to the optical simulations. A reflective coating was not

believed to be necessary after initial testing outside of the reactor, however, when the

aluminum foil covering was applied to the plates the amount of light incident on the

sensor increased nearly 125%. A possible coating would be a dielectric mirror coating

from Evaporated Coatings Inc. It is the author’s recommendation that a reflective

surface be applied to the surface closest to the reactor wall and not to the side of the

plate. By coating the back surface of the plate, the system will mimic the simulation

results to a greater degree. From observation of the functioning system, it is believed

that leaving the sides exposed is beneficial to the overall function of the system by

providing more and larger light zones.

Another change to the components should be a solid color fiber coupler to

attach to the light plates. The clear plastic was used for the prototype so that the

interface of the fibers and the light plates could be observed. However, this allowed a

substantial amount of light to escape before entering the light plates, Figure 54. A lot of

the light at the interface of the fibers and plate did not enter the plate but instead

reflected through the coupler. By using a solid white plastic such as a polyester or

acrylic instead of the clear acrylic, nearly all the light can be focused into the light plates.

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Figure 54. Light loss of from coupler at interface with light plate

Another solution to this problem could be to make the light plates slightly wider

and drill holes into the top of the plates and insert the fiber optics directly into the

plate, Figure 55. With this setup more light will enter the light plates since the tips are

embedded in the plate itself. The light that would have reflected out will have a high

chance of being refracted into the plate, either through the base of the hole or the walls

of the hole, and working down through the plate. With the modifications given here it is

believed that the reactor will be more capable of achieving the objectives laid out in

section 1.7. Again, a reflective coating could also be applied near the top of the plate on

all sides to contain more light in the plate.

Figure 55. Alternative method of interfacing fibers with light plates by embedding fibers

into the plates

Fiber optics

Light plate

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The first change should be the implementation of the white coupler. Drilling

holes into acrylic can difficult and the inside of the holes must be clear of any marks.

The glazing process used on the exterior of the light plates will probably not work on the

holes as the heat will not reach the bottom of the holes before damage is done to the

top surface of the plate. Therefore, Figure 55 should only be attempted if the solid

white coupler does not provide enough light control.

These changes to the fiber coupler component will also affect the light

distribution inside the light plate. The method used for this work may have produced

bright and dim spots in the light plates corresponding to fiber location instead of the

even distribution that was expected. By making these modifications to the coupler, the

light will be distributed much more evenly and should eliminate, or significantly

decrease the effect or, bright spots.

6.2 Future Work

A long term testing phase is recommended now that the system has proven it

can function properly. This work was involved in the design and construction only and

therefore was not concerned with long term growth. But for this system to be a

potential bioreactor for algae, the long term capabilities must be known. Growth rates

should be studied using varying CO2 content and light cycles over periods of weeks and

months. For long term testing, the reactor also needs to be modified so that the reactor

can drain from the bottom. The prototype was developed without continual growth

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considered and for long term growth studies the reactor will need to be harvested

during testing based on turbidity measurements.

Developing a controller and sensor to change the light and CO2 levels in real time

would be very helpful to prevent over or under saturation. This could be done by

attaching CO2, pH, and light sensors to MATLAB and use SimuLink and a feedback loop

attached to an injection system and the lighting system. Using an active response

system would provide optimal growth conditions at all times which would be important

in a full scale up.

A method of determining the benefit of the light plate design, or any light guide,

is to use photodiodes to measure light intensities and distributions. Photodiodes (PDs)

are the main components in commercial light sensors and are available through most

electric vendors. A box made of clear acrylic should be constructed with dimensions of

6”x3”x18” with PDs positioned around the outside of the box. The box will have to be

filled with water or even an alga solution as the light patterns are different for wet and

dry conditions. If possible, bubbling should also be done but this could be difficult in a

small volume to create the same bubble characteristics that would occur in a larger

reactor. The light guides can be inserted into the box and light passed through it. The

PDs can be run to a computer and a visual representation of the light intensity in the

box can be shown on a screen. The more PDs used, the greater accuracy the images will

have. Therefore, the entire surface of the box should be covered with PDs. This will

show the benefits of the light plates over single fibers in terms of optimal light usage.

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Long term testing should also include better understanding of the absorption of

chlorophyll a. Sample should be taken with a known chlorophyll a concentration and

the light attenuation should be measured and this data set should be used in

simulations. To do this a tool such as the spectronic 20D from Thermo Scientific

(Thermo Scientific) is needed. This device measures absorptance of a sample and thus,

knowing the chlorophyll a concentration, a complete set of absorption coefficients can

be determined. This data set would be much more accurate for the system in use and

could be used for improvements to the light system design.

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APPENDIX A: Growth material

The growth media for the experiment was a 6:1 ratio of Botanicare Pro Grow and Botanicare

Cal-Mag.

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APPENDIX B: Additional plots

5.5

5.7

5.9

6.1

6.3

6.5

6.7

6.9

7.1

7.3

0 5 10 15 20 25

pH

Test number

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5

6

7

8

9

10

11

12

22 24 26 28 30 32 34 36 38 40 42

Lig

ht

inte

nsi

ty (

µE

inst

ein

s m

-2s-1

)

distance from the top of reator(cm)

13.6

13.1

14.2

15.1

15.4

14.6

16.6

16.6

17.3

18.5

20.1

19.9

21.3

21.3

23.8

22.8

22

20.8

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APPENDIX C: Cost

Total cost of project

Item quantity units

unit

cost total cost

$ $

8" clear pvc 3.5 feet $44.50 $155.75

hardware

3/4"-10 bolts 8 unit $9.85 $78.80

3/4"-10 nuts 8 unit $0.46 $3.68

miscellaneous

hardware

$20.00

plastic

components

acrylic

12"x12"x1/4" 2 unit $14.60 $29.20

acrylic

24"x24"x1/4" 2 unit $46.20 $92.40

polyester

12"x12"1/4" 2 unit $8.03 $16.06

polyester tube

ID 3/8" OD

1/2" 6 feet $0.90 $5.40

polyester rod

1/2" 7 feet $1.95 $13.65

Flanges

blind flange 2 unit $136.00 $272.00

open flange 2 unit $123.00 $246.00

gasket 1 unit $11.00 $11.00

Sparger 1 unit $50.00 $50.00

man hours

CNC machinist 6 hours $20.00 $120.00

general shop 4 hours $10.00 $40.00

total $1,153.94

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