Chapter 1&2 Introduction & Centrifugation
Transcript of Chapter 1&2 Introduction & Centrifugation
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ASSESSMENTS :
5 QUIZZES 10%
2 ASSIGNMENTS 20%
MID SEM EXAM 30%
FINAL EXAM 40%
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CHAPTER 1 INTRODUCTION
CHAPTER 2 CENTRIFUGATION
CHAPTER 3 CHROMATOGRAPHY
CHAPTER 4 ELECTROCHEMISTRY
CHAPTER 5 MICROSCOPE
CHAPTER 6 SPECTROMETRYCHAPTER 7 ELECTROPHORESIS
CHAPTER 8 HEMATOLOGY ANALYSIS
CHAPTER 9 IMMUNOCHEMISTRY ANALYSIS
CHAPTER 10 FORENSIC ANALYSIS
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CHAPTER 1 : INTRODUCTION
BIOMEDICAL INSTRUMENTATION
Devices that can be used to make measurements of biologic or
medical quantities and give quantitative results
BASIC BIOMEDICAL INSTRUMENT
PHYSIOLOGIC SENSOR PROCESSOR DISPLAY OBSERVER
SYSTEM STORAGE
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FUNDAMENTAL RULES OF BIOMEDICALINSTRUMENTATION
Minimum disturbance to physiologic
Sensor must be at physiologic variable value
Maintain simplicity
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IMPORTANT INSTRUMENTATION TERMS
SENSITIVITY - Change in output as a function of achange in input
STABILITY
Consistency in output for a constantinput
SPECIFICITY Ability to distinguish desired variable
from other competing variables
ACCURACY the closeness of the instrumentoutput to the true value of the measured quantity
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PRECISION The smallest change in a variables thatcan be correctly measured
RESOLUTION The degree of fineness with which ameasurement can be made
REPRODUCIBILITY Same output for the same input
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TRANSFER CHARACTERISTIC
SENSITIVITY- change in output as a function of a changein input
Instrument
Output
Variable Measured
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STABILITY Consistency in output for a constantinput
Variable
Time
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SPECIFICITY Ability to distinguish desired variablefrom other competing variables
Instrument
Output
Variable Measured
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ACCURACY the closeness of the instrument output tothe true value of the measured quantity
Instrument
Output
Variable Measured
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ACCURACY
The accuracy of the instruments can be measured by usingthis calculation:
Percentage of true value = measured value true value x 100true value
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ACCURACY vs PRECISION
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POSSIBLE TYPES OF TRANSDUCERS
To convert the input signal to a form which can beconveniently processed
Mechanical Thermal
Electrical Acoustic
Mechanical Hydraulic
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TYPES OF SENSOR
PHYSICAL-THERMOMETER
BIOANALYTICAL
-GLUCOSE SENSOR
CHEMICAL-OXYGEN
ELECTRODE
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SINGLE CONVERSION SENSOR
PHYSIOLOGIC SENSOR ELECTRICAL
VARIABLE SIGNAL
PARTIAL CLARK ELECTRICAL
PRESSURE ELECTRODE CURRENT
OF OXYGEN
BODY THERMISTOR ELECTRICAL TEMPERATURERESISTANCE
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MULTIPLE CONVERSION SENSOR
PHYSIOLOGIC INTERMEDIATE SENSOR ELECTRICAL
SENSOR VARIABLE SIGNAL
BLOOD DIAPHRAGM DISPLACEMENT ELECTRICAL
PRESSURE DISPLACEMENT SENSOR SIGNAL
SERUM HYDROGEN H2O2 SENSOR ELECTRICAL
GLUCOSE PEROXIDE SIGNAL
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EXAMPLES OF FAMILIAR BIOMEDICALINSTRUMENTATION
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STANDARDS OF MEASUREMENTS
-Defined as the physical representation of the unit ofmeasurement (S.I. unit)
-For example : kilogram was originally defined as the
mass of a cubic decimetre of water at its temperature ofmaximum density at 4 C
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CALIBRATION
-Is the act or result of quantitative comparison between
a known standard and the output of the measuring
system measuring the same quantity
-Calibration curve is a references standards withknown values to cover the range of interest
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ERROR
-Defined as the differences between the measured and thetrue value
ERROR
systematic/cumulative Random error
error - arises from our
- can in principle be ability to makediscovered and corrected physical measurement
- always present and
cannot be corrected
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Systematic/cumulative Error
Instrument Environmental Zero error
Error Error
Loading
Error
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Instrument error
-Poor design of instrument-Can be avoided by selecting a suitable instrument
for a given application and calibrating instrument
using a suitable standard
Environmental error
-The condition of environment such as windy, rainy,hot .
-Can be avoided by doing the measurement in close
room.
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CHAPTER 2 : CENTRIFUGATION
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CENTRIFUGATION
-Device for separating particles from a solutionaccording to their size, shape, density,viscosity ofthe medium and rotor speed-Centrifuge have been used since middle 1800-Hand-driven (only after 1912, it is electricallydriven)
THEORY OF CENTRIFUGATION
-Sedimentation of a molecule is influenced by :a) properties of the molecule (size,shape,density)b) properties of the solvent (density, temperature)c) interactions between solute molecules & solvent
material
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FORCES IN CENTRIFUGATION
As the rotor spins in a centrifuge, centrifugal force isapplied to each molecule in the sample :
CF = Mw2r
M = mass (particle weight/molecular weight)
w = angular velocity
r = distance from the axis of rotation
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-The larger the molecule or the faster thecentrifugation or the longer the axis of rotation,
the greater the centrifugal force and the rate ofsedimentation
-Two forces act to counteract the centrifugal force are
bouyant force and frictional force
-A sedimenting molecule moves faster and faster in a
centrifugal field until :
Centrifugal force = bouyant force + frictional force
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TYPES OF CENTRIFUGES
Desktop clinical centrifuges
-Simplest & least expensive
-Max speed of most desktop centrifuges is below 3000 rpm
-Most operate at all temperature-Typical application : rapid sedimentation of blood samples
Highspeed centrifuges
-Operating up to speeds of 20,000 to
25,000 rpm
-Typical application : collect larger organelles
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Ultracentrifuges
-Typical application : separation of the major
lipoprotein fractions from plasma
-4 principles :
a)Drive and speed control : flexible drive shaft system,an overspeed system
b)Temperature control : infrared temp. sensor
c)Vacuum system
d)Rotor : swinging bucket rotor & fixed
angle rotor
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TYPES OF CENTRIFUGATION
-Analytical-Preparative
ANALYTICAL
-measuring physical properties of sedimenting particlesuch as molecular weight
PREPARATIVE-To isolate specific particles which can be reused
-Three types : rate zonal, differential, isopycnic
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Analytical
Uses small sample (less than 1 ml)
Built in optical system to analyze progress of molecules during
centrifugation Uses relatively pure sample
Used to precisely determine sedimentation coefficient and MWof molecules
Preparative
Larger sample size can be used
No optical read out collect fractions and analyze them after therun
Less pure sample can be used Can be used to estimate sedimentation coefficient and MW
Generally used to separate organelles and molecules. Mostcentrifugation work done using preparative ultracentrifuge
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Density gradient centrifugation
1) Rate zonal centrifugation
-Separates molecules based on molecular weight-Used to separate various types of macromolecules suchas different types of RNA, DNA
2) Isopycnic centrifugation-separate molecules based on density
-used to separate cell organelles
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Differential centrifugation
-Start with a uniform mixture of sample
-Centrifuge to obtain pellet of heavier molecules andsupernatant
-Take supernatant & recentrifuge to pellet the next
largest particle-Commonly used for separation of organelles
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CLASSIFICATION OF CENTRIFUGATION BASED ONRATE/SPEED
Ultracentrifugation - x> 20,000 rpm
Super speed ultracentrifugation 10,000
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ROTOR
-Two types of rotor : swinging-bucket rotor andfixed-angle rotor
SWINGING-BUCKET ROTOR
Consist of a rotor from which
hang three to six free moving
bucket that hang vertically when
the rotor is at rest
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ASSIGNMENT 1
TITLE : INTRODUCTION TO LABORATORY
INSTRUMENTS
Font : 11, Arial, 1.5 spacing
Pages : Maximum 10
Cover : Green (Name, ID Num., Lecturers name,Group, Dateline)
Dateline : 06 JULY 2011
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SAMPLE
INSTRUMENTATIONSGS 1102
ASSIGNMENT 1
INTRODUCTION TO LABORATORY INSTRUMENTS
NAME :MATRIC NO :LECTURER :GROUP :SUBMISSION DATE :