Anda di halaman 1dari 9

The AutoAnalyzer is an automated analyzer using a flow technique called continuous

flow analysis (CFA), first made by the Technicon Corporation. The instrument was invented
1957 by Leonard Skeggs, PhD and commercialized by Jack Whitehead's Technicon
Corporation. The first applications were for clinical analysis, but methods for industrial
analysis soon followed. The design is based on separating a continuously flowing stream with
air bubbles.
Operating principle
In continuous flow analysis (CFA) a continuous stream of material is divided by air bubbles
into discrete segments in which chemical reactions occur. The continuous stream of liquid
samples and reagents are combined and transported in tubing and mixing coils. The tubing
passes the samples from one apparatus to the other with each apparatus performing different
functions, such as distillation, dialysis, extraction, ion exchange, heating, incubation, and
subsequent recording of a signal. An essential principle of the system is the introduction of air
bubbles. The air bubbles segment each sample into discrete packets and act as a barrier
between packets to prevent cross contamination as they travel down the length of the tubing.
The air bubbles also assist mixing by creating turbulent flow (bolus flow), and provide
operators with a quick and easy check of the flow characteristics of the liquid. Samples and
standards are treated in an exactly identical manner as they travel the length of the tubing,
eliminating the necessity of a steady state signal, however, since the presence of bubbles
create an almost square wave profile, bringing the system to steady state does not
significantly decrease throughput ( third generation CFA analyzers average 90 or more
samples per hour) and is desirable in that steady state signals (chemical equilibrium) are more
accurate and reproducible.[1]
A continuous flow analyzer (CFA) consists of different modules including a sampler, pump,
mixing coils, optional sample treatments (dialysis, distillation, heating, etc.), a detector, and
data generator. Most continuous flow analyzers depend on color reactions using a flow
through photometer, however, also methods have been developed that use ISE, flame
photometry, ICAP, fluorometry, and so forth.
Flow injection analyzer
Flow injection analysis (FIA), was introduced in 1975 by Ruzicka and Hansen, [2] The first
generation of FIA technology, termed flow injection (FI), was inspired by the AutoAnalyzer
technique invented by Skeggs in early 1950s.[3][4] While Skeggs' AutoAnalyzer uses air
segmentation to separate a flowing stream into numerous discrete segments to establish a
long train of individual samples moving through a flow channel, FIA systems separate each
sample from subsequent sample with a carrier reagent. While the AutoAnalyzer mixes sample
homogeneously with reagents, in all FIA techniques sample and reagents are merged to form
a concentration gradient that yields analysis results.
FIA methods can be used for both fast reactions as well as slow reactions. For slow reactions,
a heater is often utilized. The reaction does not need to reach completion since all samples
and standards are given the same period to react. For typical assays commonly measured with
FIA (e.g., nitrite, nitrate, ammonia, phosphate) it is not uncommon to have a throughput of
60-120 samples per hour.
FIA methods are limited by the amount of time necessary to obtain a measurable signal since
travel time through the tubing tends to broaden peaks to the point where samples can merge
with each other. As a general rule, FIA methods should not be used if an adequate signal
cannot be obtained within two minutes, and preferably less than one. [citation needed] Reactions that
need longer reaction times should be segmented. However,considering the number of FIA

publications and wide variety of uses of FIA for serial assays, the "one minute" time
limitation does not seem to be a serious limitation for most real life assays. [citation needed] Yet,
assays based on slow chemical reactions have to be carried either in stopped flow mode
( SIA) or by segmenting the flow.
OI Analytical, in its gas diffusion amperometric total cyanide method, uses a segmented flow
injection analysis technique that allows reaction times of up to 10 minutes by flow injection
analysis.[5]
Technicon experimented with FIA long before it was championed by Ruzicka and Hansen.
Andres Ferrari reported that analysis was possible without bubbles if flow rates were
increased and tubing diameters decreased.[6] In fact, Skegg's first attempts at the auto analyzer
did not segment. Technicon chose to not pursue FIA because it increased reagent
consumption and the cost of analysis.[citation needed]
The second generation of the FIA technique, called sequential injection analysis (SIA), was
conceived in 1990 by Ruzicka and Marshal, and has been further developed and miniaturized
over the course of the following decade.[citation needed] It uses flow programming instead of the
continuous flow regime (as used by CFA and FIA), that allows the flow rate and flow
direction to be tailored to the need of individual steps of analytical protocol. Reactants are
mixed by flow reversals and a measurement is carried out while the reaction mixture is
arrested within the detector by stepping the flow. Microminiaturized chromatography is
carried out on microcolumns that are automatically renewed by microfluidic manipulations.
The discrete pumping and metering of microliter sample and reagent volumes used in SI only
generates waste per each sample injection. The enormous volume of FI and SI literature
documents the versatility of FI and SI and their usefulness for routine assays (in soil, water,
environmental, biochemical and biotechnological assays) has demonstrated their potential to
be used as a versatile research tool.
Dialyzer module
In medical testing applications and industrial samples with high concentrations or interfering
material, there is often a dialyzer module in the instrument in which the analyte permeates
through a dialysis membrane into a separate flow path going on to further analysis. The
purpose of a dialyzer is to separate the analyte from interfering substances such as protein,
whose large molecules do not go through the dialysis membrane but go to a separate waste
stream. The reagents, sample and reagent volumes, flow rates, and other aspects of the
instrument analysis depend on which analyte is being measured. The autoanalyzer is also a
very small machine
Recording of results
Previously a chart recorder and more recently a data logger or personal computer records the
detector output as a function of time so that each sample output appears as a peak whose
height depends on the analyte level in the sample.
Commercialization
Technicon sold its business to Revlon in 1980 [7] who later sold the company to separate
clinical (Bayer) and industrial (Bran+Luebbe - now SEAL Analytical) buyers in 1987. At the
time, industrial applications accounted for about 20% of CFA machines sold.
In 1974 Ruzicka and Hansen carried out in Denmark and in Brasil initial experiments on a
competitive technique, that they termed flow injection analysis (FIA). Since then the
technique found world wide use in research and routine applications, and was further

modified through miniaturization and by replacing continuous flow with computer controlled
programmable flow.
During the 1960s industrial laboratories were hesitant to use the autoanalyzer. Acceptance by
regulatory agencies eventually came about by demonstration that the techniques are no
different from a recording spectrophotometer with reagents and samples added at the exact
chemical ratios as traditionally accepted manual methods.[8]
The best known of Technicon's CFA instruments are the AutoAnalyzer II (introduced 1970),
the Sequential Multiple Analyzer (SMA, 1969), and the Sequential Multiple Analyzer with
Computer (SMAC, 1974). The Autoanalyzer II (AAII) is the instrument that most EPA
methods were written on and reference.[citation needed] The AAII is a second generation segmented
flow analyzer that uses 2 millimeter ID glass tubing and pumps reagent at flow rates of 2 - 3
milliliters per minute. Typical sample throughput for the AAII is 30 - 60 samples per hour. [9]
Third generation segmented flow analyzers were proposed in the literature, [10] but not
developed commercially until Alpkem introduced the RFA 300 in 1984. The RFA 300 pumps
at flow rates less than 1 milliliter per minute through 1 millimeter ID glass mixing coils.
Throughput on the RFA can approach 360 samples per hour, but averages closer to 90
samples per hour on most environmental tests. In 1986, Technicon (Bran+Luebbe) introduced
its own microflow TRAACS-800 system.[11]
Bran+Luebbe continued to manufacture the AutoAnalyzer II and TRAACS, a micro-flow
analyzer for environmental and other samples, introduced the AutoAnalyzer 3 in 1997 and the
QuAAtro in 2004. The Bran+Luebbe CFA business was bought by SEAL Analytical in 2006
and they continue to manufacture, sell and support the AutoAnalyzer II/3 and QuAAtro CFA
systems, as well as Discrete Analyzers.
And there are other manufacturers of CFA instruments.
Skalar Inc., subsidiary of Skalar Analytical, founded in 1965, which has its head office in
Breda (NL), is since its founding an independent company, fully owned by its personnel.
Development in robotic analyzers, TOC and TN equipment, and monitors has extended the
product lines of its long life SAN++ Continuous Flow Analyzers. Software packages for data
acquisition and analyzer control are also in house products, running with latest software
demands and handles all analyzer hardware combinations.
Astoria-Pacific International, for example, was founded in 1990 by Raymond Pavitt, who
previously owned Alpkem. Based in Clackamas, Oregon, U.S.A., Astoria-Pacific
manufactures its own micro-flow systems. Its products include the Astoria Analyzer lines for
Environmental and Industrial applications; the SPOTCHECK Analyzer for Neonatal
screening; and FASPac (Flow Analysis Software Package) for data acquisition and computer
interface.
FIAlab Instruments, Inc., in Bellevue Washington, also manufactures several analyzer
systems.
Alpkem was purchased by Perstorp, and then later by OI Analytical in College Station Texas.
OI Analytical manufactures the only segmented flow analyzer that uses polymeric tubing in
place of glass mixing coils. OI is also the only major instrument manufacturer that provides
segmented flow analysis (SFA) and flow injection analysis (FIA) options on the same
platform.

Clinical analysis
AutoAnalyzers were used mainly for routine repetitive medical laboratory analyses, but they
had been replaced during the last years more and more by discrete working systems which
allow lower reagent consumption. These instruments typically determine levels of albumin,
alkaline phosphatase, aspartate transaminase (AST), blood urea nitrogen, bilirubin, calcium,
cholesterol, creatinine, glucose, inorganic phosphorus, proteins, and uric acid in blood serum
or other bodily samples. AutoAnalyzers automate repetitive sample analysis steps which
would otherwise be done manually by a technician, for such medical tests as the ones
mentioned previously. This way, an AutoAnalyzer can analyze hundreds of samples every
day with one operating technician. Early AutoAnalyzer instruments each tested multiple
samples sequentially for individual analytes. Later model AutoAnalyzers such as the SMAC
tested for multiple analytes simultaneously in the samples.
In 1959 a competitive system of analysis
Specialties Company. That system became
represented by an instrument known as the
Sample Analysis method slowly replaced
laboratory.

was introduced by Hans Baruch of Research


known as Discrete Sample Analysis and was
"Robot Chemist." Over the years the Discrete
the Continuous Flow system in the clinical

Industrial analysis
The first industrial applications - mainly for water, soil extracts and fertilizer - used the same
hardware and techniques as clinical methods, but from the mid-1970s special techniques and
modules were developed so that by 1990 it was possible to perform solvent extraction,
distillation, on-line filtration and UV digestion in the continuously flowing stream. In 2005
about two thirds of systems sold worldwide were for water analysis of all kinds, ranging from
sub-ppb levels of nutrients in seawater to much higher levels in waste water; other common
applications are for soil, plant, tobacco, food, fertilizer and wine analysis.
Current Uses
AutoAnalyzers are still used for a few clinical applications such as neonatal screening or
Anti-D, but the majority of instruments are now used for industrial and environmental work.
Standardized methods published by the ASTM (ASTM International), the US Environmental
Protection Agency (EPA) as well as the International Organization for Standardization (ISO)
for environmental analytes such as nitrite, nitrate, ammonia, cyanide, and phenol.
Autoanalyzers are also commonly used in soil testing laboratories, fertilizer analysis, process
control, seawater analysis, air contaminants, and tobacco leaf analysis.

P
Roche Cobas 6000

Roche Cobas u 411

Chemistry analysers: Access (left); Coulter (right).

Racks: for putting samples, quality controls or calibrations. Cobas 6000

An automated analyser is a medical laboratory instrument designed to measure different


chemicals and other characteristics in a number of biological samples quickly, with minimal
human assistance.
Many methods of introducing samples into the analyser have been invented. This can involve
placing test tubes of sample into racks, which can be moved along a track, or inserting tubes
into circular carousels that rotate to make the sample available. Some analysers require
samples to be transferred to sample cups. However, the effort to protect the health and safety
of laboratory staff has prompted many manufacturers to develop analysers that feature closed
tube sampling, preventing workers from direct exposure to samples.]
Samples can be processed singly, in batches, or continuously.
The automation of laboratory testing does not remove the need for human expertise (results
must still be evaluated by medical technologists and other qualified clinical laboratory
professionals), but it does ease concerns about error reduction, staffing concerns, and safety.
Routine biochemistry analysers
These are machines that process a large portion of the samples going into a hospital or private
medical laboratory. Automation of the testing process has reduced testing time for many
analytes from days to minutes. The history of discrete sample analysis for the clinical
laboratory began with the introduction of the "Robot Chemist" invented by Hans Baruch and
introduced commercially in 1959[1].
AutoAnalyzer is an automated analyzer using a special flow technique named "continuous
flow analysis (CFA)", invented in 1957 by Leonard Skeggs, PhD and first made by the
Technicon Corporation. The first applications were for clinical (medical) analysis. The
AutoAnalyzer profoundly changed the character of the chemical testing laboratory by
allowing significant increases in the numbers of samples that could be processed. The design
based on separating a continuously flowing stream with air bubbles largely reduced slow,
clumsy, and error prone manual methods of analysis.
The types of tests required include enzyme levels (such as many of the liver function tests),
ion levels (e.g. sodium and potassium, and other tell-tale chemicals (such as glucose, serum
albumin, or creatinine).
Simple ions are often measured with ion selective electrodes, which let one type of ion
through, and measure voltage differences.[3] Enzymes may be measured by the rate they
change one coloured substance to another; in these tests, the results for enzymes are given as
an activity, not as a concentration of the enzyme. Other tests use colorimetric changes to
determine the concentration of the chemical in question. Turbidity may also be measured.
Immuno-based analysers
Antibodies are used by some analysers to detect many substances by immunoassay and other
reactions that employ the use of antibody-antigen reactions.
When concentration of these compounds is too low to cause a measurable increase in
turbidity when bound to antibody, more specialised methods must be used.

Recent developments include automation for the immunohaematology lab, also known as
transfusion medicine.

Hematology analysers
These are used to perform complete blood counts, erythrocyte sedimentation rates (ESRs), or
coagulation tests.
Cell counters
Automated cell counters sample the blood, and quantify, classify, and describe cell
populations using both electrical and optical techniques. Electrical analysis involves passing a
dilute solution of the blood through an aperture across which an electrical current is flowing.
The passage of cells through the current changes the impedance between the terminals (the
Coulter principle).[4] A lytic reagent is added to the blood solution to selectively lyse the red
cells (RBCs), leaving only white cells (WBCs), and platelets intact. Then the solution is
passed through a second detector. This allows the counts of RBCs, WBCs, and platelets to be
obtained. The platelet count is easily separated from the WBC count by the smaller
impedance spikes they produce in the detector due to their lower cell volumes.
Optical detection may be utilised to gain a differential count of the populations of white cell
types. A dilute suspension of cells is passed through a flow cell, which passes cells one at a
time through a capillary tube past a laser beam. The reflectance, transmission and scattering
of light from each cell is analysed by sophisticated software giving a numerical
representation of the likely overall distribution of cell populations.
Some of the latest hematology instruments may report Cell Population Data that consist in
Leukocyte morphological information that may be used for flagging Cell abnormalities that
trigger the suspect of some diseases.
Reticulocyte counts can now be performed by many analysers, giving an alternative to timeconsuming manual counts. Many automated reticulocyte counts, like their manual
counterparts, employ the use of a supravital dye such as new methylene blue to stain the red
cells containing reticulin prior to counting.[5] Some analysers have a modular slide maker
which is able to both produce a blood film of consistent quality and stain the film, which is
then reviewed by a medical laboratory professional.
Coagulometers
Automated coagulation machines or Coagulometers measure the ability of blood to clot by
performing any of several types of tests including Partial thromboplastin times, Prothrombin
times (and the calculated INRs commonly used for therapeutic evaluation), Lupus
anticoagulant screens, D dimer assays, and factor assays.
Coagulometers require blood samples that have been drawn in tubes containing sodium
citrate as an anticoagulant. These are used because the mechanism behind the anticoagulant
effect of sodium citrate is reversible. Depending on the test, different substances can be added
to the blood plasma to trigger a clotting reaction. The progress of clotting may be monitored
optically by measuring the absorbance of a particular wavelength of light by the sample and
how it changes over time.

Other hematology apparatus


StaRRsed Inversa, automated Westergren-based ESR 'analyzer'
Automatic erythrocyte sedimentation rate (ESR) readers, while not strictly analysers, do
preferably have to comply to the 2011-published CLSI (Clinical and Laboratory Standards
Institute) "Procedures for the Erythrocyte Sedimentation Rate Test: H02-A5 and to the ICSH
(International Council for Standardization in Haematology) published "ICSH review of the
measurement of the erythrocyte sedimentation rate", both indicating the only reference
method, being Westergren, explicitly indicating the use of diluted blood (with sodium
citrate), in 200 mm pipettes, bore 2.55 mm. After 30 or 60 minutes being in a vertical
position, with no draughts and vibration or direct sunlight allowed, an optical reader
determines how far the red cells have fallen by detecting the level.
Miscellaneous analysers
Some tests and test categories are unique in their mechanism or scope, and require a separate
analyser for only a few tests, or even for only one test. Other tests are esoteric in nature
they are performed less frequently than other tests, and are generally more expensive and
time-consuming to perform. Even so, the current shortage of qualified clinical laboratory
professionals has spurred manufacturers to develop automated systems for even these rarely
performed tests.
Analysers that fall into this category include instruments that perform:

DNA labeling and detection

Osmolarity and osmolality measurement

Measurement of glycosylated haemoglobin (haemoglobin A1C), and

Aliquotting and routing of samples throughout the laboratory

Anda mungkin juga menyukai