Automation in clinical pathology

By Bhanupriya
• Automation is a process of replacement of
tasks normally performed by humans with
computerized methods
• Automation in clinical laboratory is a process
by which analytical instruments perform many
tests with least involvement of an analyst
 Advantages
• Increased efficiency and speed.
• Accuracy and precision.
• Multiple tests on a single platform.
• Reduction of labor costs
• Improved turn around time
• Reduction in sample volumes needed for
analysis
 Disadvantages
• Expensive and high running costs.
• Space occupying machinery.
• Discordance of results between different
analysers
 Automation in urine
• Automated instruments in urinalysis include:
Individual strip readers
Semiautomated analysers
Fully automated chemistry analysers
Automated urine cell analysers
Completely automated systems
• Automated reagent strip readers – use
spectrophotometric measurement of light
reflection termed REFLECTANCE PHOTOMETRY
• Light reflection from test pads decreases in
proportion to intensity of colour produced by
concentration of test substance
• A monochromatic light is directed towards
reagent pads by placing a filter between light
source and reflective surface of the pad
• The light is reflected to a photodetector and
an analog / digital converter
• The instruments compare the amount of light
reflection with that of known concentrations,
then display or print concentration units
• Semi automated instruments are still dependant
on a operator for specimen mixing , test strip
dipping
• Fully automated chemistry analysers add urine to
the reagent strip and automated urine cell
analysers mix, aspirate , dilute and stain urine to
classify urine sediment particles
• Automated urine systems perform a complete
urinalysis that includes physical , chemical, and
microscopic parts of routine urinalysis
 EQUIPMENT MANUFACTURER
WAIVED URINE CHEMISTRY INSTRUMENTS

Clinitek 50 Siemens

Clinitek status Siemens

Chemstrip 101 Roche

Ursys 1100 system Roche

SEMIAUTOMATED CHEMISTRY INSTRUMENTS

Clinitek 200/200+ Siemens

Clinitek 500 Siemens

Chemstrip criterion II Roche

Chemstrip urine analyser Roche

Urisys 1800 system Roche

Miditron junior II Roche

FULLY AUTOMATED CHEMISTRY
INSTRUMENTS
Clinitek atlas Siemens

Chemstrip super automated urine Roche

analyser
Urisys 2400 system Roche

AUTOMATED MICROSCOPY

UF-100 urine cell analyser Sysmex

iQ200 automated urine microscopy Iris

AUTOMATED URINALYSIS SYSTEMS

ADVIA urinalysis workcell Siemens

iQ200 automated urinalysis system Iris

 Clinitek 50/100

• It is suited for small Lab

• Reagent strips are manually dipped and placed into

the strip reader

• Results are displayed or printed

• Patients ID, specimen color, clarity are manually

entered

• Abnormal results are flagged

Basic principle of automated urine
analysers

Electrical impedance

Image-based analysis
Most common urine analyser
 Manual vs automatic
• Manual microscopic examination requires well
trained and experienced staff and consumes a
considerable amount of time.
• Automatic urine sediment analyzers for high-
volume laboratories were developed in order
to provide better standardization, improve the
certainty of measurement and save staff time.
 Iris iQ200 ELITE analyzer
• Analytical principle: Digital Flow Morphology
technology using the AutoParticle Recognition
(APR) software.
• As the urine passes through the flow cell, urine is
illuminated by a special light source, and the
images are recorded by a digital camera placed
into the eyepiece of the microscope and
transmitted to the computer. The software
classifies these images and displays them on the
screen for the operator for final decision.
 The system Classifies urine particles in the photographs

based on size, shape, texture and contrast in to

following categories – RBCs, WBCs, WBC clumps,

hyaline casts, unclassified casts, squamous and non

Squamous epithelial cells, bacteria, yeast, crystals,

mucus
 Siemens Clinitek Microalbumin 2
Reagent Strips
• Provide albumin, creatinine, and albumin to
creatinine ratio results in 1 minute

• useful to test for microalbuminuria in patients with

diabetes or hypertension in order to detect early
kidney disease
 Siemens Clinitek Microalbumin 9
Reagent Strips
• Provide albumin, blood, creatinine, glucose,
ketone, leukocyte, nitrite, pH, & protein and
albumin to creatinine ratio & protein to
creatinine ratio
 The Chemstrip Super Automated Urine Analyser

and the Roche Diagnostics Urisys 2400 system are

fully automated ‘walk-away’ urine chemistry

instruments for a large urinalysis laboratory.

 With the Chemstrip Super Automated Urine

Analyser, sample volumes are detected, adjusted

and automatically mixed.

• A sorter mechanism supplies a single test

strip from the sorter drum to a sorter

position.

• A gripping mechanism grasps the test strip

and dips it in to the urine specimen tube.

• The dipping mechanism lifts the test strip out of

the sample tube while removing excess urine by

dragging the strip along the inside of the

specimen tube

• The dipping mechanism then transfers the test

strip to the Reflectance Photometer position.

• A transport plate positions the test strip at the

Reflectance Photometer recording head, where

specimen is measured at three different

wavelengths (555,620,660 nm) at 48 seconds and

120 seconds after dipping.

• The Urisys 2400 system utilizes a pippetting unit that

automatically mixes the specimen and pipettes the

precise volume to each test pad.

• The minimum sample volume is 1.5 ml. Four hundred

test strips are loaded into a Urisys 2400 cassette and

the strips are stable in the cassette for 2 weeks

 Automated Microscopy
• Automated Urine cell analyzers provide efficient

standardized results in about a minute, markedly

improving turnaround times.

• The Sysmex UF – Series offers fully automated sample

analysis with automatic classification of all formed

element groups with scattergrams and histograms for

reference
Disadvantages of Manual Urine Sediment
Microscopy

o Subjective element identification

o Poor reproducibility
o Lack of standardization
o Time consuming/labor intensive
• The UF-100 and UF –50 use laser – based flow

cytometry along with impedance detection,

forward light scatter and fluorescence to identify

the individual characteristics and stained urine

sediment particles in a flowing stream.

SYSMEX UF 50 SYSMEX UF100
• The sample is stained with two dyes that
radiate an orange and green fluorescence. The
DNA within the cells is stained by the orange
dye, phenathridine ; the nuclear membranes,
mitochondria and negatively charged cell
membranes are stained with a green dye,
carbocyanine
The stained sample is passed through the flow

cell, presented to a laser light beam that

produces fluorescence and light scatter

• Particles are identified by measuring the
change in impedance of the sediment
elements, as well as the height and width of
the fluorescent and light scatter signals,
presented in scattergrams and histograms
• The main parameters are RBCs, WBCs,
epithelial cells, casts and bacteria. Flagging
parameters include pathologic casts, crystals,
small round cells, sperm and yeast like cells
 Automated Urinalysis Systems

• Clinitek Atlas, an automated urine chemistry analyzer

and the Sysmex UF-100, an automated urine cell

analyzer, have been integrated to develop the ADVIA

Urinalysis Workcell System.

 LAURA M urine analyser by ERBA
• 600 measurements / hour
• Measurement Principle – reflectance
photometer
Parameters measured:
Blood
Leukocytes
• Bilirubin
• Urobilinogen
• Ketones
• Glucose
• Proteins
• PH
• Nitrites
• Specific gravity
• Microalbuminuria
 CASA
• CASA provides objective determination of
sperm velocity and direction of motion
• Sperm concentration and morphology are also
included
• Currently, CASA instrumentation is found
primarily in labs that specialise in andrology
and perform a high volume of semen analyses
• MES developed SQA-VISION
• High resolution visual interface
• Bar code scanning technology
• Built-in LIS capability
• Touch-to-Mark Technology for assessing both
sperm Vitality and conducting full differential
Morphology.
• Required sample size is 0.5 ml but there is an
option for even 0.25 ml
• Even 15 microl option for IVF and sperm
banking
• provides a precise and accurate semen
analysis in just 75 seconds reporting sperm
count, morphology, motility and other
additional parameters
Sperm Concentration
% Motility expressed as total and a,b,c and d
Rapid Progressive Motility (a)
Slow Progressive Motility (b)
Non-progressive Motility (c)
Immotility (d)
% Normal Morphology
Motile Sperm Concentration
Progressively Motile Sperm Concentration
Functional Sperm Concentration
Average Velocity
Sperm Motility Index
 Why Computer assisted semen
analysis
• Accuracy and reliability
• Easy to carry out
• Objective data
• Data storage for further reference
• The results are reproducible and consistent
• No much variability in results between
laboratories
 Requirement of CASA
• To ensure indifference in semen count ,
motility and morphology estimates
• To reduce interpersonal variations and to
standardize observables with higher limits of
acceptance
• Reliable reproducibility
 BODY FLUIDS
• The Sysmex XE-5000 is the first fully
automated, multi-parameter hematology
analyzer to have a dedicated body fluid mode.
• The analyzer is easily and quickly switched
between peripheral blood mode and body
fluid mode by a keypad touch.
• Once converted to body fluid mode, the
analyzer automatically performs a
background cell count.
• The XE-5000 body fluid application employs
the same fluorescent flow technology used for
peripheral blood analysis but provides
extended linearity for low cell count body
fluids through a unique extended cell counting
process.
• The analyzer aspirates 130 μL of sample (same
as peripheral blood mode) but has an
extended count cycle for the body fluid
application.
• This allows the instrument to analyze 3 times
the volume of sample normally analyzed.
• The XE-5000 body fluid application provides a
WBC, a total nucleated cell count (TC), RBC
count
• While the WBC count includes those nucleated cells
that fall in the usual counting area for peripheral
WBCs, the TC includes all nucleated cells and is the
more commonly reported result for body fluid cell
counts.
• The nucleated cells analyzed in the body fluid WBC
channel are subcategorized as polymorphonuclear
and mononuclear, providing a rapid 2-part
differential analysis.
• All parameters are reportable to 3-decimal places.
• Scattergram and histogram displays allow
interpretation of results and identification of
interfering particles
• Evaluation of the XE-5000 body fluid
application in a large clinical laboratory
showed it to be an accurate, sensitive, and
efficient method for analyzing cerebrospinal
fluid (CSF), serous, and synovial fluid cell
counts.
• Of critical importance when performing body
fluid cell counts on automated analyzers is the
analyzer’s ability to detect and flag non-
cellular substances and particles that may
interfere with the accuracy of the results.
• It is not uncommon to encounter debris,
crystals, infectious microorganisms, abnormal
proteins, or lipids in various fluids.
• Sysmex recently launched the XN-Series, which
contains an improved BF module compared to the XE-
5000
• RBCs are counted in the RBC channel using sheath flow
impedance technology
• WBC-mononuclear cells and PMN cells are
determined by flow cytometry in the differential
channel.
• The differential channel combines forward scatter (size
of the cell) with side scatter (inner complexity of the
cell) and the fluorescence intensity (DNA/RNA content)
to identify and cluster each cell.
• This channel also reports high fluorescence
body fluid cells (HF-BF), such as macrophages
and mesothelial cells.
• These HF-BF cells are found just above the MN
cluster and are not included in the WBC
(differential) count, but are included in the total
nucleated cell count .
• If the number of HF-BF cells present exceeds a
preset cutoff value, the “ WBC abnormal
scattergram ” flag will be generated.
• This cut-off value is user-definable.
• Additionally, if non-cellular particles, such as
bacteria or cell debris are present, leading to
erroneous clusters, the “ WBC abnormal
scattergram ” flag will be automatically
generated
• Furthermore, the XN-1000 BF module has
several new features compared to the XE-5000
BF mode.
• It is able to count 4 times more WBCs than
the XE-5000, aspirates less sample volume (88
μ L) compared to the XE-5000 (130 μ L) and
offers open or closed tube sampling in fluids
and blood as well as a micro-tube adapter for
easy handling of pediatric tubes.
Thank you