AUTOANALYZER

BIOSENSORS
• Is a chemical sensor having optical device or transducer and a
biological recognition element.
• The concentration of the analyte is recognized by an enzyme based
biosensor or an affinity based biosensor.
• They help in estimation of urea,glucose,creatinine.
• Implantable subcutaneous glucose sensors are also being used.
• Intravascular sensors that relrases nitric oxide to prevent thrombosis.
AUTOANALYZER
• it is the ultra modern ,highly sophisticated equipment
in which Large number of samples can be analyzed
within a short span.
• Automation is mechanization of various duties like
labelling, separating, pipetting, mixing, incubating,
calculating printing and preparing QC charts.
• Autoanalyzer will take care of the increased work load
in the laboratories by automation of machines.
 TYPES OF ANALYSER
SEMIAUTOANALYSER FULLY AUTOMATED ANALYSER
• In semiautomated analyser some of • In fully automated analyser all the
the steps like pipetting of steps for analysis are done
sample,reagent mixing,and incubation automatically.
are manually done.while
measurement ,calculation and • More accurate.
printing of results are automatically
done. • More cost .
• Less accurate. • Require more space.
• Less cost. • Less time is needed.
• Less space.
• More time is needed.
• Analyse only one parameter at a time.
Semiautoanalyzer
BECKMAN COULTER
PARTS OF AUTOANALYZER
1. Breaks & fuses
2. Power supply unit
3. Water supply/drain unit
4. Mixing unit
5. Sample transfer unit
6. Syringe unit
7. Transfer unit
8. Wash nozzle unit
UNIT OPERATION IN AN ANALYTICAL PROCESS
1. Specimen identification
2. Specimen delivery
3. Specimen processing
4. Sample introduction and internal transport
5. Sample loading and aspiration
6. Reagent handling and storage
7. Reagent delivery
8. Chemical reaction phase
9. Measurement approaches
10. Signal processing , data handling, and process control.
10.Measurement approaches
• Photometry/spectrophotometry
• Reflectance photometry
• Fluorometry
• Turbidimetry and nephelometry
• Chemiluminiscence and bioluminescence
• Electrochemical
11.Signal processing ,data handling and process control: done
automatically by the computer software and results are transfer online.
TYPES OF AUTOANALYZER
• CONTINOUS FLOW ANALZYER
• DISCRETE AUTOANALYZER :BATCH ANALYZER AND RANDOM ACCESS
ANALYZER
• CENTRIFUGAL ANALYZER
• DRY CHEMISTRY ANALYZER
CONTINOUS FLOW ANALYZER
• It is the first autoanalyzer discovered. In these systems sample and
reagent are passed sequentially through the same analytical pathway
• Advantage: possible to test large number of specimens for a
particular test accurately and precisely
• Disadvantage: the machine does not allow test selection: all test must
be performed even if not requested.
• Chances of carry over will be there i.e., transfer of a quantity of
analyte or reagent from one specimen reaction into a subsequent one
present.
• High cost of maintanence.
Discrete autoanalyzer
This type of autoanalyzer are sample oriented, have the capacity of
analysing simultaneously many parameters in a single sample,
They are the most versatile analyzers used in laboratory. They are of
two types:
1. Batch analyzer performs only one type of test at a time and
provision for only one type of reagent at a time, hence they are
parameter oriented
2. Random access analyzer are the most common autoanalyzer used
in laboratory ,they complete all tests on one sample before
proceeding to the next sample
CENTRIFUGAL ANALYZER
• In this, analysis is based on centrifugal force to add simultaneously
reagents with sample and displace reaction mixture in hollows equally
distributed on outer border of a rotor

• Detection by spectrophotometer

• Advantage :

rapid batch analysis of sample at a time.

Low maintenance with low sample and reagent consumption and

Low cost per test

• Disadvantage : only one test type can be performed each time.

DRY CHEMISTRY ANALYZER
• Here all reagents necessary for the reaction are embedded on a
plastic matrix in their dry state.
• Reaction is initiated by the addition of the serum over the matrix and
the color of reaction is measured by reflectance spectrophotometry.
• This method is very useful for emergency sample of critical care and
point of care tests eg: glucometer used for diabetic patients for blood
glucose analysis
ADVANTAGES OF AUTOANALYZER
1. Large numbers of samples can be tested in a short time
2. Decreases turn over time(TAT)
3. Variety of tests can be performed by using various methods such as
end point and kinetic reaction.
4. Methods performed on autoanalyzer are accurate, precise, sensitive
and specific
5. It performs more work with less technical staff.
6. Quality control programs both Internal and external can be
implemented efficiently and effectively by using autoanalyzers.
7. It is very safe from biohazardous material as there is minimum
contact with the specimen and reagents.
 ADVANCEMENTS
• ROBOTIC AUTOMATION is advance form of fully automated analyser
uses automated guided vehicles called mobile robots to transfer
sample within or outside lab
.
COLORIMETER
 Introduction
• Colorimeter is a light sensitive instrument that measures how much
light is absorbed by a colored compound.
• Colorimeter is the laboratory instrument which works on the
principle discussed below:
• PRINCIPLE:
• Coloured solutions have the property of absorbing light of definite
wave length. When a beam of monochromatic light is passed
through a coloured solution ,the coloured
substance{chromophore}absorbs some amount of light and the rest
is transmitted.
• The absorption of light is related to the colour intensity which is
proportional to the concentration of the substance that is
responsible for producing the colour.
• The quantum of light that is absorbed by the coloured solution is
determined by the optical instrument called COLORIMETER or
PHOTOMETER.
• LAWS OF COLORIMETRY:
• The two fundamental laws which express quantitatively the extent of
light absorption are:
 Beer’s law

• According to this law ,the amount of light transmitted through

a coloured solution decreases exponentially with increase in
concentration of the coloured substance i.e,absorbing
molecule.
• It indicates that the amount of light absorbed is directly
proportional to the concentration of coloured particles in the
solution.
 Lambert’s law

• According to this law,the transmitted light decreases exponentially

with an increase in the thickness of the layer of solution through
which the light passes.it indicates that the amount of light absorbed
in the coloured solution is directly proportional to depth/distance
through which light passes in the solution.
• In colorimeter, the length/depth/distance through which light passes
is kept constant by using test tubes or cuvettes of the same
diameter for both test and standard so that only variable is
concentration.
• Transmittance of light [t]:
• The proportion of light that passes through is called transmittance.
• It is usually expressed as a percentage of maximum light transmitted when
the coloured solute is absent.
• T = I/Io
• %T= I/Io x 100
• I = transmitted light
• I0 = incident light
• The relation between the amount of light transmitted and the concentration
of solute is inversely proportional and logarithmically
• i=io ect
• i=intensity of transmitted light
• io =intensity of incident light
• e=molar extinction coefficient
• c=concentration of absorbing substance
• t= thickness of medium
• Absorbance or Optical density:
• It is the amount of light absorbed by coloured solute, so directly
proportional to the concentration of solute.

• The relation between transmittance and the absorbance is given by

the following formula :
• A = 2– log% T
• A = ecl
• Where e= molar absorptivity
• C= concentration of the analyte
• l = path length or thickness of the medium
 COMPONENTS OF COLORIMETER

1.Source of light: light source for measurement of visible

portion of light spectrum is electric[tungsten] lamp for
wavelength of 320 to 700 nm.
Other lamps may be used like deuterium for UV light
2.Slit: It is adjustable and allows only a beam of light to
pass through it.it will exclude unwanted stray light.
3.Filter or Monochromator:filters are used for selecting
light of single wave length i.e, monochromatic light. Filters
will absorb light of unwanted wave lengthand allow only
required monochromatic light to pass through.The wave
length which is maximally absorbed for a particular colour is
the required wave length.Filters are made of glass or dyed
gelatin.
Colour of the Wave length in nm Filter/complementa
solution ry colour
violet 350-430 Yellow blue
blue 430-475 yellow
Green blue 475-495 orange
Blue green 495-505 red
Green 505-555 purple
Yellow green 555-575 violet
yellow 575-600 blue
orange 600-650 Green blue
red 650-700 Blue green
4.Cuvette or sample holder: They are special type of tubes made of
optical glass to hold the coloured sample for the measurement in the
colorimeter.
a) The cuvette used in a particular colorimeter should be optically
matched i.e, glass cuvette are used to view visible light range.
Quartz or silica for UV range
b) For accurate and precise reading cuvette must be transparent,
clean and devoid of any scratches and there should be no bubble
adhering to inner surface of filled cuvette.
c) The cuvette may be square or rectangle or test tube like but the
optical path is always 1cm
• 5.detector or photo cell:it is used to convert the light
energy in to electrical energy.Photo cell is a metal plate
which is coated with layer of photo sensitive element such
as selenium or cadmium.This is positive electrode.This
light sensitive material is covered with a semi transparent
thin layer of gold/copper. when light strikes the selenium
layer the electrons are liberated which pass into the
transparent layer making it electronegative.The potential
difference [electric current]thus generated is directly
proportional to the light intensity striking the photo cell.
6.Read out device[measuring device]:electrical
energy from a detector is displayed by read out
device. the current generated is generally calibrated
to read directly as transmittance or absorbance or
both.
PROCEDURE
• In clinical lab the serum sample and the reagent are mixed
thoroughly and incubated at 370 degrees to develop colour
optically after which the OD is determined and the
concentration of the substance is calculated.this is called END
POINT ANALYSIS .
• The serum and the reagent are incubated and readings are
taken at 2 and 3 minutes exactly and the difference in OD
between the 2 values is used to calculayte the concentration in
test samples.this is KINETIC ANALYSIS.here optimum colour
is not developed but it is quicker and hence is often used in
auto analyser.
• Let A1 be the absorbance of test solution ,C1 its concentration
• Let A2 be the absorbance of standard solution and C2 be its concentration

• A1∝ C1 A1 =K C1,

• A2∝ C2 A2 =K C2

• A1÷A2 =C1÷C2
• If this value of concentration of test solution present in x ml of sample taken then 100 ml
of sample contains
• C1=[ A1÷A2 ]×[C2÷x ] ×100
 Applications of colorimeter

• Used in hospitals and laboratories for Estimation of

biochemical compounds in Blood, Plasma, Serum, CSF,
Urine and other body fluids
• The following biochemical compounds can be estimated
colorimetrically
Glucose, Urea, Creatinine, Lipids, proteins and bilirubin etc.
• Used in food, paint and textile industry
• To test water quality by screening for Cl, Fl,CN etc
ADVANTAGES DISADVANTAGES
1. Inexpensive 1. It cannot be used for colourless
compounds
2. Easily transportable
2. Doesn’t work in UV and IR region
3. Very well applicable for quantitative
estimation 3. Can’t set specific wavelength as we
have to set a range
4. Similar colours from interfering
substances can produce error in
results
 Spectrophotometer
• It is an instrument which has all the basic components of a colorimeter with more
sophistication
• It works on the principle of beer –Lambert’s law
• It measures analyte concentration in colored and colorless solution
• Wavelengths of UV & IR are also utilized here Unlike in colorimeter where only
visible region is utilized
• Here light is separated into continuous spectrum of wave length and passed through
solution using prism or diffraction grating unlike in colorimeter where filters are used
and wavelength of one color are grouped together
• Cuvettes are made of silica or Quartz as glass absorbs UV wave length
 Applications

1. Qualitative & Quantitative determinations such as enzyme

assays, molecular weight determination
2. In analytical chemistry quantitative determination of different
analytes, Metal Ions, Biological Macro molecules
3. Spectroscopic analysis is commonly studied in solutions but
solids, gases may also be studied
4. Useful in pharmaceutical , food industry , paints industry
ADVANTAGES DIS ADVANTAGES
1. Can be used for colourless 1. More expensive
compounds 2. Not transportable
2. Works in UV,VISIBLE and IR regions
3. Specific wave length can be set
4. More accurate
Thank you
QUALITY CONTROL
 QUALITY CONTROL
• Quality control is an integral part of a clinical laboratory.
• The purpose of QC is the reliability of each measurement performed
on a sample
• Quality is defined as conformance to satisfying the needs and
expectation of the customers
• If the errors are promptly detected corrective action can be taken.
• Quality control is of two types:
1.External Quality control
2.Internal Quality control
EXTERNAL QUALITY CONTROL INTERNAL QUALITY CONTROL

External quality control is a method Internal quality control to monitor and

that allows for comparison of a ensure the reliability of test results
laboratory’s testing to a source outside produced by the laboratory.
the laboratory.
External quality control
scheme(EQAS) compares the
performance of different laboratories.
Done once or twice in month. Done on a daily basis.
This is necessary for maintaining long It is necessary for maintaining accuracy
term accuracy of analytical method. & precision of analytical method on
daily basis.
TRUE VALUE AND MEASURED VALUE

TRUE VALUE MEASURED VALUE

Refers to the actual concentration of Refers to the value of the

the analyte being measured. concentration of the analyte that you
get of experiment.
 ACCURACY AND PRECISION
ACCURACY PRECISION
Refers to the closeness of a result to the Refers to the repeatability and
true value. reproducibility of a test.

It indicates how well a measurement It indicates how well a series of

agrees with an accepted value. measurements agree with each
 SPECIFICITY AND SENSITIVITY
• Specificity
SPECIFICITY
of a reaction denotes • Sensitivity
SENSITIVITY
of an assay is the
that only one substance will fraction of those with a disease
answerofthat
Specificity particular
a reaction test.
denotes that only that assay
Sensitivity correctly
of an predicts.
assay is the fraction of
• Is
one substance will answer
the ability that particular
of an assay to • It iswith
those theaability
disease of
thata assay
methodcorrectly
to
test.determine the concentration of predicts.
detect the analyte without giving
the target analyte without false negatives.
influence from potentially
Is the ability of an
interfering assay to determine
substances the
or factors It is the ability of a method to detect the
concentration of thematrix.
in the sample target analyte without analyte without giving false negatives.
influence from potentially interfering
substances or factors in the sample matrix.
TOTAL QUALITY MANAGEMENT
• CALIBRATOR is the material of known qualitative and/or quantitative
characteristics used to calibrate a measurement procedure or to compare the
response obtained with the response of a test specimen.

• CONTROL MATERIAL is a device, solution or pooled specimen to be used

in the QC process.

• REFERENCE MATERIAL is a substance having homogenous and well

established values for the calibration of an apparatus, an assay method, or
assigning values to material.
 LABORATORY ERROR

• A laboratory error is a defect occurring at any part of the laboratory

cycle from ordering tests to reporting results and appropriately
interpreting and reacting on these.
 SOURCES OF ERRORS
1.PREANALYTICAL ERRORS:
• errors that occur from the time of specimen collection to the time right
before it is processed.
• Incorrect identification of patient
• Improper preparation of patient
• Improper collection of blood sample
• Incorrect labelling
• Incorrect specimen storage
• Transport error
• Incorrect specimen container
• Incorrect test request
• 2.ANALYTICAL ERRORS:
• errors that occur during the actual testing of the specimen.
• Instrument maintenance
• Calibration and request errors
• Improper measurement of specimen or reagent
• Wrong reagent preparation
• Control is out of range
• Incorrect analytical procedure
3.POSTANALYTICAL ERRORS:
• errors of transcription, validation, delays in the reporting or incorrect
reference values.
• Transcription errors in reporting
• Diluting errors-incorrect dilution or dilution factors
• Lack of training
• Report not sent
• Report sent to the wrong location
• Wrong date entry
• Wrong report
 RANDOM AND SYSTEMATIC ERRORS
RANDOM ERRORS SYSTEMATIC ERRORS
Errors that arise from limitations of Errors caused by a defect in the
physical measurements. analytical method or by an
improperly functioning
instrument
indeterminate errors. determinate errors.
 QUALITY CONTROL CHARTS
• These are used to compare the observed control value with control limits.

• They provide a visual display that can be quickly reviewed.

• Control charts help to detect accuracy problems(shift in mean) and precision

problems(shit in SD).

• The values will indicate if the analytical run is in control(acceptable) or out of

control(unacceptable).
 LEVEY-JENNINGS CHARTS
• Levey Jennings charts are a graphical method for displaying control results and
evaluating whether a procedure is in control or out of control.
• On Levey Jennings chart, the solid line in the middle represent mean .
• On x axis -days of month and on y axis the values of repeat control measurement
is shown.
• SD intervals are represented by dash lines.
WESTGARD RULES
• For internal quality control we use the Westgard rules which are plotted on
Levy-Jennings charts.
• They are used to identify the type of errors.
• Westgard rules:
• 1-2s rule
• 2-2s rule
• 1-3s rule
• 4-1s rule
• R-4s rule
• 10x rule
 ADVANTAGES OF WESTGARDS
MULTI RULE SYSTEM
• The main advantage by multi rule system is distinguishing between
random and systematic error.
• They also indicate the direction of a investigation of a systematic error
should take.
THANK YOU!