ASSESSMENTS :

5 QUIZZES 2 ASSIGNMENTS MID SEM EXAM FINAL EXAM

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CHAPTER 1 CHAPTER 2 CHAPTER 3 CHAPTER 4 CHAPTER 5 CHAPTER 6 CHAPTER 7 CHAPTER 8 CHAPTER 9 CHAPTER 10

INTRODUCTION CENTRIFUGATION CHROMATOGRAPHY ELECTROCHEMISTRY MICROSCOPE SPECTROMETRY ELECTROPHORESIS HEMATOLOGY ANALYSIS IMMUNOCHEMISTRY ANALYSIS FORENSIC ANALYSIS

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 SYSTEM SENSOR PROCESSOR DISPLAY STORAGE OBSERVER

FUNDAMENTAL RULES OF BIOMEDICAL INSTRUMENTATION

Minimum disturbance to physiologic Sensor must be at physiologic variable value Maintain simplicity

IMPORTANT INSTRUMENTATION TERMS SENSITIVITY - Change in output as a function of a change in input STABILITY Consistency in output for a constant input SPECIFICITY Ability to distinguish desired variable from other competing variables ACCURACY the closeness of the instrument output to the true value of the measured quantity

PRECISION The smallest change in a variables that can be correctly measured RESOLUTION The degree of fineness with which a measurement can be made
REPRODUCIBILITY Same output for the same input

TRANSFER CHARACTERISTIC
SENSITIVITY- change in output as a function of a change in input

Instrument Output

Variable Measured

STABILITY Consistency in output for a constant input

Variable

Time

SPECIFICITY Ability to distinguish desired variable from other competing variables

Instrument Output

Variable Measured

ACCURACY the closeness of the instrument output to the true value of the measured quantity Instrument Output

Variable Measured

ACCURACY
The accuracy of the instruments can be measured by using this calculation:
Percentage of true value = measured value true value x 100 true value

ACCURACY vs PRECISION

POSSIBLE TYPES OF TRANSDUCERS To convert the input signal to a form which can be conveniently processed Mechanical Electrical Mechanical Thermal Acoustic Hydraulic

TYPES OF SENSOR

PHYSICAL -THERMOMETER

CHEMICAL -OXYGEN ELECTRODE

BIOANALYTICAL -GLUCOSE SENSOR

SINGLE CONVERSION SENSOR PHYSIOLOGIC VARIABLE SENSOR ELECTRICAL SIGNAL

PARTIAL PRESSURE OF OXYGEN

CLARK ELECTRODE

ELECTRICAL CURRENT

BODY

THERMISTOR

ELECTRICAL TEMPERATURE RESISTANCE

MULTIPLE CONVERSION SENSOR

PHYSIOLOGIC SENSOR BLOOD PRESSURE

INTERMEDIATE VARIABLE DIAPHRAGM DISPLACEMENT

SENSOR

ELECTRICAL SIGNAL ELECTRICAL SIGNAL

DISPLACEMENT SENSOR

SERUM GLUCOSE

HYDROGEN PEROXIDE

H2O2 SENSOR

ELECTRICAL SIGNAL

EXAMPLES OF FAMILIAR BIOMEDICAL INSTRUMENTATION

STANDARDS OF MEASUREMENTS
-Defined as the physical representation of the unit of

measurement (S.I. unit) -For example : kilogram was originally defined as the mass of a cubic decimetre of water at its temperature of maximum density at 4 C

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 with

known values to cover the range of interest

ERROR
-Defined as the differences between the measured and the

true value

ERROR

systematic/cumulative error - can in principle be discovered and corrected

Random error - arises from our ability to make physical measurement - always present and cannot be corrected

Systematic/cumulative Error

Instrument Error

Environmental Error Loading Error

Zero error

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.

CHAPTER 2 : CENTRIFUGATION

CENTRIFUGATION -Device for separating particles from a solution according to their size, shape, density,viscosity of the medium and rotor speed -Centrifuge have been used since middle 1800 -Hand-driven (only after 1912, it is electrically driven) 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

FORCES IN CENTRIFUGATION As the rotor spins in a centrifuge, centrifugal force is applied to each molecule in the sample : CF = Mw2r

M = mass (particle weight/molecular weight) w = angular velocity r = distance from the axis of rotation

-The larger the molecule or the faster the centrifugation or the longer the axis of rotation, the greater the centrifugal force and the rate of sedimentation
-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

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

Ultracentrifuges -Typical application : separation of lipoprotein fractions from plasma

the major

-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

TYPES OF CENTRIFUGATION
-Analytical -Preparative

ANALYTICAL -measuring physical properties of sedimenting particle such as molecular weight PREPARATIVE -To isolate specific particles which can be reused -Three types : rate zonal, differential, isopycnic

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 MW of molecules
Preparative Larger sample size can be used No optical read out collect fractions and analyze them after the run Less pure sample can be used Can be used to estimate sedimentation coefficient and MW Generally used to separate organelles and molecules. Most centrifugation work done using preparative ultracentrifuge

Density gradient centrifugation 1) Rate zonal centrifugation -Separates molecules based on molecular weight -Used to separate various types of macromolecules such as different types of RNA, DNA 2) Isopycnic centrifugation -separate molecules based on density -used to separate cell organelles

 Differential centrifugation

-Start with a uniform mixture of sample -Centrifuge to obtain pellet of heavier molecules and supernatant -Take supernatant & recentrifuge to pellet the next largest particle -Commonly used for separation of organelles

CLASSIFICATION OF CENTRIFUGATION BASED ON RATE/SPEED Ultracentrifugation - x> 20,000 rpm Super speed ultracentrifugation 10,000 <x<20,000 rpm Low speed centrifugation x<10,000 rpm

ROTOR -Two types of rotor : swinging-bucket rotor and fixed-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

ASSIGNMENT 1
TITLE

: INTRODUCTION TO LABORATORY INSTRUMENTS : 11, Arial, 1.5 spacing : Maximum 10 : Green (Name, ID Num., Lecturers name, Group, Dateline) : 06 JULY 2011

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