14-08-2020

AUTOMATION

DR.V.G.KARPAGHAVALLI
DEPT OF BIOCHEMISTRY
STANLEY MEDICAL COLLEGE

OBJECTIVES
• Definitions
• Advantages
• History
• Basic concepts
• Steps involved in automation

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Automation
• Automation in clinical chemistry describes the process
where an analytical instrument perform many tests
with only minimal involvement of an analyst.

• The International Union of Pure and Applied Chemistry

(IUPAC) define automation as "The replacement of
human manipulative effort in the performance of a
given process by mechanical and instrumental devices
that are regulated by feedback of information so that
an apparatus is self-monitoring or self adjusting”.

Advantages
• Handle larger workloads without comparable
increases in staff.
• Instrument performs a repetitive task by itself,
• allows the instrument to perform a variety of different
tasks.
• Intelligent automation allow some individual
instruments to self-monitor and respond appropriately
to changing conditions.
• reduction in the errors of analysis through the
elimination of tasks that are repetitive .
• The improved reproducibility has led to a significant
improvement in the quality of laboratory tests.
• less time consumption per sample analysis,
• more number of tests done in less time,

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Evolution of automation
DATES SAMPLING PROCESSING REACTION COMPUTATION

1901-1950 H H H H

1950-1959 M H/M M H/M

1960-1969 M M M M/A

1970-1979 M M A A

1980 to M/A A A A
present

• H-HUMAN
• M- MECHANISATION
• A- AUTOMATION

History
• 1956- first auto analyser Leonard Skeggs –Glucose,
urea, calcium
• Multichannel system 20 analytes-150samples/hr
• 1959- Robot Chemist discrete analysis
• 1968 -centrifugal analysers for discrete analysis
• 1970 –electronic automation and LIS
• 1980 random access analysers, discrete, STAT,
endpoint ,kinetic assay, photodiode array
• Dry chemistry analysis
• Integrated modular analysers
• Total lab automation

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Terminology
• Single channel analysis
• Multiple channel analysis
• Sequential analysis
• Batch analysis
• Discrete analysis
• Throughput

Basic Concepts
• Automated analyzers generally incorporate
mechanized versions of basic manual
laboratory techniques and procedures.
• modern instrumentation is packaged in a wide
variety of configurations
• random-access analysis
– Analyses are performed on a collection of
specimens sequentially, with each specimen
analyzed for a different selection of tests.

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other configurations
• Continuous- Flow analyzers:-first automated
analyzers used in clinical laboratories.
– Single-channel analysis
– Multiple-channel analysis
• Centrifugal analyzers -use discrete pipetting
to load aliquots of specimens and reagents
sequentially into discrete chambers in a rotor,
with the specimens subsequently analyzed in
parallel .

Unit Operations in an Analytical

Process
• Specimen identification
• Specimen delivery
• Specimen processing
• Sample introduction and internal transport
• Sample loading and aspiration
• Reagent handling and storage
• Reagent delivery
• Chemical reaction phase
• Measurement approaches
• Signal processing, data handling, and process control

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1-Specimen Identification
• the identifying link between patient and
specimen is noted at the patient’s bedside,
and maintained throughout
– 1) transport of the specimen to the laboratory,
– 2) subsequent specimen analysis, and
– 3) preparation of a report

Bar code labeling

• The unique label is affixed to the specimen
collection container
• Code 39 is followed
• A bar coding system
• bar code is an array of rectangular bars and
spaces arranged in a predetermined pattern
• light beam from the scanner is absorbed by the
dark bars but reflected by the light spaces.
• light signal  electrical signal  digitized.

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Code 39 allows both the patient’s name and id no in human and machine readable form

Advantages of using bar code labels

• Elimination of work lists for the system
• Avoidance of mistakes during placement of tubes
in the analyzer
• Analysis of specimens in random sequence.
• Avoidance of possible tube mix-up when serum
must be transferred to a secondary container.
• Decrease in identification errors.
– Automatic reading of bar code labels reduces the
error rate from 1 :300 characters (for human entry) to
about 1 : million

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RFID labels
• RFID labels can be encrypted to shield the
encoded data, ensuring that sensitive
information is kept confidential.

2-Specimen Delivery
• Automated methods often used to deliver
specimens to the laboratory include:-
– pneumatic tube systems,
– electric track vehicles, and
– mobile robots.

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Pneumatic tube systems

• Rapid specimen transportation
and are reliable when installed
as point-to-point services.

Mobile robots
• used successfully to
transport laboratory
specimens both within
the laboratory and
outside the central
• Some mobile robots have
been integrated with
robotic systems that
automate loading and
unloading of specimens
• others initiate an audible
or visual signal on arrival
at a specified station

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Electric track vehicles

• Electric track vehicles have a larger carrying capacity
than pneumatic tube systems and do not have
problems with damaging specimens by acceleration
and/or deceleration forces.
• Some systems maintain the carrier in an upright
position
• The containers can hold dry ice or refrigerated gel
packs with specimens if desired.
• Disadvantage
– cost of moving the track and loading/unloading stations
– staff may be necessary to unload the carts and transport
the specimens to their final destination

3-Specimen Preparation
Conventional
• The clotting of blood in
specimen collection tubes,
• their subsequent
centrifugation, and
• the transfer of serum to
secondary tubes
• If performed manually, the
process results in a delay in
the preparation of a
specimen for analysis.
• Automation
• Use of whole blood
– For ISE
– Dry reagent films
• Automated preparation
– Special wheel 2 degree
freedom X and Z
• Use of robotics

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Basic configurations of robotic devices

CARTESIAN CYLINDRICAL ARTICULATING

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4-Sample handling, transport and delivery

• In most situations the specimen for automatic
analysis is serum or plasma.
• Analyzers sample directly from primary
collection tubes or cups with aliquots from the
original specimen tubes
• Sample cups are designed to:-
– minimize dead volume,
– minimize evaporation,
– decrease thermolability,
– photodegradation

Sample loading zone may be

• (1) a revolving tray or turntable,
• (2) a mechanical belt, or
• (3) a rack or set of racks by which specimens are
delivered in a predetermined order to the sample
aspiration station of the analyzer.
Sample probe design
• The use of level sensors, which restrict the
penetration of sample probes into specimens and
provide smoother motion control, greatly
reduces splatter.
• Developed closed-container sampling systems
– the specimen probe passes through a hollow needle
that initially penetrates the primary container’s
rubber stopper.

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Sample loading zone

5-Specimen Processing
• Removal of Protein and Other Interferents
– (1) dialysis,
– (2) column chromatography, and
– (3) Filtration
• Separations in Immunoassay Systems
To separate free and bound antibodies , automated immunoassay
analyzers use bound antibodies or proteins in a solid-phase format.
– (1) beads,
– (2) coated tubes,
– (3) microtiter plates,
– (4) magnetic and nonmagnetic microparticles, and
– (5) fiber matrices.

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Separations in Immunoassay Systems

Sample transport and delivery

Continuous-flow systems, • In discrete analysis
(Random access analysis)

peristaltic pumps Positive–liquid displacement pipettes

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Centrifugal analyzers

6-Reagent Handling and Storage

• Ready to use liquid reagents
• Inventory is maintained by system -no of tests
• On board refrigerated storage for reagents
• Some analyzers use dry tablet reagents
• Others use reagent-impregnated slides or
strips.
• Still others rely entirely on electrodes to react
with specimens

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Reagent Identifcation
• Labels on reagent containers include
information such as
– (1)reagent identification,
– (2) volume of the contents or number of tests,
– (3) expiration date, and
– (4) lot number.
• Many reagent containers carry bar codes that
contain some or all of this information,

advantages of reagent bar codes

• inventory management,
• ability to insert reagent containers in random
sequence, and
• ability to automatically dispense a particular
volume of liquid reagent.
• level sensing system on the reagent probe, alert
the operator
• In immunoassay systems, a bar code on a reagent
container contains key information about
calibration curve .

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Open Versus Closed Systems

• With an open analyzer, the operator is able to
use“in-house” reagents or reagents from a
variety of suppliers.
• Such analyzers usually have considerable
flexibility and are adapted readily to new
methods and analytes.
• A closed-system analyzer requires the reagent to
be provided by the manufacturer.
• Most immunoassay systems and point-of –care
applications are closed

7-Reagent Delivery
• Liquid reagents are delivered to mixing and
reaction chambers by pumps or by positive
displacement syringe devices.
• For those analyzers in which more than one
reagent is acquired and dispensed by the
same syringe, washing or flushing of the probe
is essential to prevent reagent carry-over.

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8-Chemical Reaction Phase-

• continuous-flow system –

• In discrete analysis (Cuvette )

Reusable cuvettes needs laundry systems for Disposable on board manufactured

rinsing and drying cuvettes

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Mixing of Reactants
• In a discrete system, these include:
– 1. Forceful dispensing.
– 2. Magnetic stirring.
– 3. Vigorous lateral displacement.
– 4. A rotating paddle.
– 5. The use of ultrasonic energy.
• Continuous-flow analyzers- tumbling action of the stream
in a mixing coil.
• Dry reagent systems -the serum completely interacts with
the dry chemicals as it flows through the matrix of the
reaction unit.

Dry reagent systems

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Timing of Reactions
• In discrete random-access analyzers, samples
and reagents are added to a cuvette in a timed
sequence, and detector signals are measured
at intervals to follow the course of each
reaction.
Thermal Regulation
• Devices used to control temperature include
– (1) air baths,
– (2) water baths, and
– (3) contact with warm plates.

9-Measurement Approaches
Automated chemistry analyzers use a variety of optical
measurement devices including
– (1) photometers
– (2) spectrophotometers,
– (3) reflectance photometers,
– (4) fluorometers, and
– (5) luminometers.
Immunoassay systems use reaction schemes that
produce
– (1) fluorescence,
– (2) chemiluminescence, and
– (3) electrochemiluminescence.
Ion-selective electrodes and other electrochemical
techniques

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Photometry/Spectrophotometry
• Measurement of absorbance requires the following three basic components
• 1. An optical source – tungsten lamps, quartz-halogen lamps, deuterium
lamps,mercury lamps, xenon lamps, and lasers. 300 to 700 nm.
• 2. A means of spectral isolation-
– interference filters peak transmissions of 30% to 80% and bandwidths of 5 to
15 nm
– Monochromators with movable gratings and slits provide a continuous choice
of wavelengths.
– many manufacturers use a stationary, holographically ruled grating, coupled
with a stationary photodiode array, to isolate the spectrum
• 3. A detector.
– Photodiodes either as individual components or in multiples as an array.
– Photomultiplier tubes
• Proper alignment of cuvets with the light path(s) is important in both
automated and manual analyzers.

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Reflectance Photometry
• The intensity of the reflected light from the
reagent carrier is compared with that reflected
from a reference surface

Fluorometry
Fluorescence is the emission of electromagnetic
radiation by a species that has absorbed exciting
radiation from an outside source.
Turbidimetry and Nephelometry
to measure plasma proteins and to perform
therapeutic drug monitoring.
Chemiluminescence and Bioluminescence
excitation event is caused by a chemical or
electrochemical reaction

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Electrochemical methods
• The most widely used in ion-selective electrodes.
• These electrodes have replaced flame
photometry in the determination of sodium and
potassium.
• The relationship between ion activity and the
concentration of ions in the specimens must be
established with calibrating solutions, and such
electrodes need to be recalibrated frequently to
compensate or alterations in electrode response.

10-Signal Processing, Data Handling, and

Process Control
• Analog signals from detectors are converted to digital forms by
analog to-digital converters at a rate of 103 to 105 conversions per
second.

• complex, nonlinear standard responses from nonisotopic

immunoassays and reflectance spectrometry are readily
transformed into linear calibration curve

• Autoverification -is the process whereby patient results generated

from interfaced instruments are compared by computer software
against laboratory-defined acceptance parameters.
– If results fall within these defined parameters, the results are
automatically released for reporting with no additional
laboratory staff intervention.
– Any data that fall outside the defined parameters are reviewed
by laboratory staff prior to reporting.

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Unit Operations in an Analytical

Process
• Specimen identification
• Specimen delivery
• Specimen processing
• Sample introduction and internal transport
• Sample loading and aspiration
• Reagent handling and storage
• Reagent delivery
• Chemical reaction phase
• Measurement approaches
• Signal processing, data handling, and process control

STAND ALONE AUTOANALYSER

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INTEGRATED MODULE CHEMISTRY ,

ISE AND IMMUNOASSAY

TOTAL AUTOMATION FOR THE

CLINICAL LABORATORY
• Significant progress has been made in
automating preanalytical and postanalytical
activities and integrating these operations
with analytical systems.

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Automated Specimen Processing

• receiving the specimen,
• inspecting it for appropriateness (labeling, container
type, temperature, and quantity of specimen),
• logging onto the LIS,
• labeling the specimen with an accession number, and
• separating urgent and stat specimens from routine
specimens.
• sort for centrifugation and aliquoted
• sort into instrument-specific racks for analyzers.

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TOTAL LAB AUTOMATION

• conveyor transport the specimens are moved to
various workstations,
• interfacing to automated analyzers,
• more sophisticated process control, and,
• a specimen storage and retrieval system.
• Some systems use an open design, which permits
interfaces to analyzers from a variety of vendors,
whereas other systems are of a closed design and are
interfaced only to the vendor’s own or a limited
number of analyzers.

Conveyor Belts- main feature of integrated or so-

called total laboratory automation systems

Loop conveyor

a unidirectional conveyor

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Laboratory Automation System (LAS)

• Also called middleware
• Is the control software that read specimen’s bar code
and obtain information from the laboratory’s LIS about
specimen type and ordered tests.
– calculate the number of aliquots and the proper volume for
each depending on the tests requested,
– route the specimens to analyzers,
– recap the specimens,
– retain the specimens for repeat, reflex, and dilution testing.
– autoverification of the test results
– linked with HMIS and results released along with
demographic details of patient and doctor’s name

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PRACTICAL CONSIDERATIONS
Automation of a part or all operations
Evaluation of Requirements:
• Evaluation begins with mapping of the current
laboratory work-flow from the arrival of patient
specimens through completion of testing and
reporting of results.
• Determining process steps that
– (1) are bottlenecks,
– (2) waste labor, and
– (3) are prone to error

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Other areas
Urine analyzers
• It is more difficult, because of the broad range of
concentrations of many urine constituents
• This requires a low limit of detection to measure
low concentrations and
• expanded linearity to permit measurements of
high concentrations without dilution.
• However, with new-generation analyzers,
automation of urinalysis is gaining acceptance

Cell Counters
complete blood count (CBC) have been
automated through the use of
• the “Coulter principle,” which is based on cell
conductivity,
• light scatter, and
• flow cytometry.

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To conclude
• The history of automation in the clinical laboratory is long
and varied.
• Manual testing is clearly of the past century for a modern
laboratory except for a few very specialised tests.
• modern automated methods have become inevitable
especially to avoid the inherent variability of manual
procedures.
• The challenge is for laboratories to embrace the right kind
of automation to best meet their specific patient testing
needs
• There are myriad options available, from stand-alone
integrated systems, to pre- and post-analytical modules
that can be mixed and matched with analytical units, to
total laboratory automation. This is definitely a situation in
which “One size does not fit all.”

Thank you

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References

1. James C. Boyd, and Charles D. Hawker, chapter 19, Automation in the

Clinical Laboratory, Tietz Textbook Of Clinical Chemistry And Molecular
Diagnostics 5th ed(2012),469-485.
2. David A Armbruster, David R Overcash, Jaime Reyes, Clinical Chemistry
Laboratory Automation in the 21st Century - Amat Victoria curam (Victory
loves careful preparation) Clin Biochem Rev 35 (3) 2014:143-153
3. Bakan Ebubekir, Ozturk Nurinnisa and Kilic-Baygutalp Nurcan* Automation
in the clinical laboratory: integration of several analytical
andintralaboratory pre- and post-analytical Systems ,Turk J Biochem 2017;
42(1): 1–13

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