Medical Lab Instrument

UNIT-3
BIO-CHEMICAL INSTRUMENT
Medical Laboratory
Instrument
SUJITH.M, LECTURER/EEE, VIDYAA VIKAS ENGG COLLEGE Ph.No.
9600286256
Equipment
Spectrophotometer
Is optical device that measure light absorption at various
wavelengths for a given liquid sample.
Blood cell analyzer
Is a device to measures the number of red and white
blood cells per scaled volume.
The aperture impedance and flow cytometery
sujith.m,lecturer/eee,9600286256
Equipment
Glassware, centrifuges, suction devices
Colorimeter
Is an optical devise that measures the color concentration
of a substance in solution
Flame photometer
Is an optical electronic devise that measures the color
intensity of substance that have been aspirated into a
flame (sodium and potassium)
Equipment
Ph/ blood gas analyzer
Is a device which measure blood Ph, Po2, Pco2
Chromatograph and Autoanalyzer
Is a electromechanical device used to separate, identify,
and measure the concentration of substances in a liquid
medium.
Computer based record and operation system
Electrophoresis
Electrophoresis is the motion of dispersed particle
relative to a fluid under the influence of a spatially
uniform electric field
This electro-kinetic phenomenon was observed for

the first time in 1807 by Reuss
There are several types of electrophoresis, but
the concepts are similar.
The machine has an anode (positive charge)
and a cathode (negative charge).
Negative ions move toward the anode, and
positive-charged ions move towards the
cathode.
The rate and distance traveled by these
molecules help scientists classify and study
different bimolecular.
What is Gel Electrophoresis?
Electro = flow of electricity,
phoresis, from the Greek = to carry across
A gel is a colloid, a suspension of tiny particles in a
medium, occurring in a solid form, like gelatin

Gel electrophoresis refers to the separation of charged
particles located in a gel when an electric current is

applied
Charged particles can include DNA, amino acids, peptides,
etcsujith.m,lecturer/eee,9600286256
Principles of Gel Electrophoresis
Electrophoresis is a technique used
to separate and sometimes purify
macromolecules - especially
proteins and nucleic acids - that
differ in size, charge or
conformation
 When charged molecules are
placed in an electric field, they
migrate toward either the positive or
negative pole according to their
charge. In contrast to proteins,
which can have either a net positive
or net negative charge, nucleic acids
have a consistent negative charge
imparted by their phosphate
backbone, and migrate toward the
anode.
Definition
Separation of DNA fragments according
to size, based on movement through a gel
medium when an electric field is applied.
DNA negatively charged
DNA cut up using restriction endonucleases
Make up the gel which the DNA will be put
into
Dye added to the DNA
Buffer solution added to the tank
DNA samples loaded into wells
Electrical current applied to the chamber
DNA is stained using ethidium bromide
Colorimeter
The colorimeter is a filter photometer that measures
the color concentration of a substance in solution, this
is accomplished electronically by detecting the color
light intensity passing through a sample containing the
reaction products of the original substance and reagent
Eg: urine test
Basic colorimeter analysis
 Involves the precise measurement of light intensity.
Transmittance is defined as:
T= (I1/I0) x 100%
I2= T I1
I2=T2 I0
I0 is initial light intensity

I1 is first attenuated light intensity
I2 is second attenuated light intensity
T is the transmittance in percent
Absorbance is defined as:
A= log(I0/I1)=log 1/T
Beer’s law
A= aCL
L is cuvette path length
C is concentration of absorbing substance
a is absorbtivity (~ substance)
Concentration of unknown solution
Cμ= Cs Aμ/As
Cμ is unknown concentration
Cs standard concentration
Aμ is unknown absorbance
As is standard absorbance
Calibration
1. Ground the amplifier input V1 and adjust
potentiometer R4 and adjust potentiometer R4 for
0V ± 5mV at the amplifier output.
2. Remove the ground and place reference
concentration in cuvettes 1 and 2
3. Adjust potentiometer (R1) for 0V ± 10mv at the
amplifier output.
Calibration
4. Leave the reference concentration in cuvette 1 and
replace cuvette 2 with a cuvette containing the
sample.
5. Read the unbalanced voltage on the meter in
percentage of transmittance or absorbance units.
Calibration
Example: if V1 = +1 mV with reference cuvettes
and V1 = +25 mV with reference and sample
cuvette, R2 = 2 kΩ, R3 = 1
kΩ
Calculate the voltage read on the meter display for
both conditions of V1.
Solution: Av = 1+ R2/R3 = 1+ 2/1 = 3
Av = Vm/V1
Condition 1: Vm = Av * V1 = 3 * 1mV = 3mV
Condition 2: Vm = Av * V1 = 3 * 25 mV = 75 mV
Maintenance
Calibration
Adjustment
Replacement of burned-out lamp and photo detector
Electronic problems are rare
Medical Laboratory Instrumentation
Spectrophotometer
Definition, Purpose, Use
Spectrophotometer measures light absorption by a
liquid substance at various wavelengths.
The Components of unknown material can be
determined, or the concentration of a number of
known substances can be measured.
Eg: blood cell resistivity
Operation, Calibration
A monochromator uses a diffraction grating or prism to
disperse the light from the lamp (slit S1).
The light is broken into its spectral components as it
arises from slit S2 and falls on the sample in the cuvette.
Narrower slits give rise to shorter wavelenghts.
The angle of the diffraction grating determines light
wavelength if all other parameter are fixed.
The mirror reduces equipment size
Light output, Photodetector sensitivity, and sample
substance absorption change with wavelengths
measurement
Schematic
Maintenance
Calibration
Adjustment
Replacement of light source bulbs
Replacement of the Photodetector
Mechanically rotating assemblies (mirrors,…)
Electronics faulty
Flame photometer
It is measure the color intensity of a flame
that is supported by oxygen and a specific
substances
The basic schematic shows that a reference
gas containing a lithium salt causes a red light
to shine on the reference photodetector
through the optical filter
A yellow or violet light from sample sodium
or potassium falls on the sample photodetector
It is similar to colorimeter
sujith.m,lecturer/eee,9600286256
Continuous calibration can be accomplished by
inspiration of air( oxygen to support
combustion) and lithium.
The output read in units of sodium or
potassium concentration
Maintenance
Adjustment of bulbs & photo detectors
Aspiration device and flame chambers
occasionally cleaning
Electronics failure usually frequent
Auto analyzer
Purpose of Autoanalyzer
The autoanalyzer is sequentially measures
blood chemistry and displays this on a graphic readout
This is accomplished by
– Mixing
– reagent
– Reaction
– Colorimetric measurement in continuous stream
Elements of Autoanalyzer
Sampler
Proportioning pump and manifold
 Dialyzer
 Heating bath
 Colorimeter
 Record
Sampler
Aspirates samples, standards, and wash solutions to the
autoanalyzer system
Proportioning pump
Introduces samples with reagents to effect the proper chemical
color reaction to be read by the colorimeter.
 Pumps fluids at precise flow rates to other modules, as proper
color development depends on reaction time and temperature
Dialyzer
 Separates interfacing substances from the sample material by
permitting selective passage of sample components through a
semipermeable membrane
Heating bath
Heats fluids continuously to exact temperature (37 degree).
Colorimeter
 Monitors the changes in optical density of the fluid stream
flowing through a tubular flow cell.
 Color intensities proportional to substance concentrations
 Colorimeter convert the color intensity to equivalent
electrical voltages
Recorder
Converts the electrical signal from the colorimeter into a
graphic display on moving chart
Maintenance
 Calibration and adjustment
Mechanical
– Tubing
– Moving pump parts
 Electrical
– Switches
– Motors
Electronic failures are few Problems
 Identification of samples
 Sterilization for sample and glassware and
equipment parts

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UNIT-3

This electro-kinetic phenomenon was observed for

Electro = flow of electricity,

phoresis, from the Greek = to carry across

A gel is a colloid, a suspension of tiny particles in a

medium, occurring in a solid form, like gelatin

particles located in a gel when an electric current is

Eg: urine test

I0 is initial light intensity

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