Distributed Sensor Network
description
Transcript of Distributed Sensor Network
![Page 1: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/1.jpg)
Distributed Sensor Network
Graham Wiley
Leonid Sukharnikov
![Page 2: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/2.jpg)
Arabinose Sensor
HSL Amplifier
Levels of Abstraction - System
![Page 3: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/3.jpg)
Arabinose Promoter
HSL Sender/
Reciever
HSL Sender
CFP YFP
Levels of Abstraction - Devices
![Page 4: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/4.jpg)
Levels of Abstraction - Parts
![Page 5: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/5.jpg)
Expected Output
Expanding YFP ring surrounding CFP circle shows signal propogation
Media plate w/ bacterial lawn
![Page 6: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/6.jpg)
Arabinose
Mechanism
![Page 7: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/7.jpg)
HSL
Mechanism
![Page 8: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/8.jpg)
HSLHSL
Mechanism
![Page 9: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/9.jpg)
Construction
21
3
4 parts
Final
![Page 10: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/10.jpg)
Construction Results
21
34 parts
Final
![Page 11: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/11.jpg)
FUTURE WORK
• Check construct via sequencing• Reconstruct if necessary
![Page 12: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/12.jpg)
Simone Macmil
Durga P Sarvepalli
4 November 2006
Detecting small molecule signaling using phosphorylation dependent
mechanism in E.coli
![Page 13: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/13.jpg)
SMALL MOLECULES
• Smallest part of pure chemical substance that retains
its structure and properties.
• Play an important role in multiple signaling
mechanisms
![Page 14: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/14.jpg)
APPLICATIONS OF SMALL MOLECULES IN BIOLOGY
• Small molecule drugs
• Biopolymers
• Synthetic peptides
• Primers
![Page 15: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/15.jpg)
NEED FOR SMALL MOLECULE DETECTION
• Understand molecule – protein interactions
• Effect of molecules on the viability of cells
• Genetic changes caused by molecules used in molecular
therapy
• Drug discovery
![Page 16: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/16.jpg)
CURRENTLY AVAILABLE TECHNIQUES
• Analytical: NMR, Western blot, Spectrophotometry, Chromatography, ELISA
• Small molecule – protein interactions : Nanowire sensors ( Wang et al , PNAS
2005;102;3208-3212)
• Disadvantages: whole cells cannot be used
• Using micro channels whole cells can be used and less amount of reagents
required
• Micro channels can also be customized according to the design of the
experiment and allows studies to be conducted under flow
![Page 17: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/17.jpg)
Glass chrome Mask
PhotoresistSubstrate
Exposure to UV Light
Pattern template
PDMS with channel
PDMS
FABRICATION OF MICROFLUIDIC CHANNEL
PDMS - Poly dimethyl siloxane
![Page 18: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/18.jpg)
MICROFLUIDIC CHANNEL DIMENSIONS
Channel width : 200µm depth : 50 µm
![Page 19: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/19.jpg)
DEVICE
Induction
OmpRLacI
Aspartate detector
Tar- EnvZ receptor EYFP
J04500 C0082 B0015 I6110
![Page 20: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/20.jpg)
METHODS
Transformation
Parts from the registry
Restriction enzyme digestion
Sequential ligation to create construct
Sequence transformants
Ligate into pUC
Grow selected transformants in micro channels
![Page 21: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/21.jpg)
RESULTSAspartate detection
Microfluidic channel
Cells grown in Minimal Media lacking aspartate and induced with IPTG
Cells fluoresce after 30 min in the presence of LB
![Page 22: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/22.jpg)
Movement of E.coli in the channel
RESULTS
![Page 23: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/23.jpg)
FUTURE WORK
• Improvise microchannels
- Study flow of bacteria under various conditions - chemotaxis, sensitivity of bacteria in the gut to pH, temperature.
• Detect low concentration of small molecules formed during bio-industrial production.
-Design appropriate sensors for detection (small peptides and sugars)
![Page 24: Distributed Sensor Network](https://reader035.fdocuments.net/reader035/viewer/2022062803/56814740550346895db47ca5/html5/thumbnails/24.jpg)
ACKNOWLEDGEMENTS
Dr. Bruce Roe
Dr. Matthias Nollert
Dr. David Schmidtke
Dr. Fares Nazar
Dr. Randall Hewes
Doug White
Sandra Bryant