In clinical chemistry, automation is the mechanization of the steps in a procedure. Manufacturers design the instruments to mimic manual techniques.

1957 introduction of the first automated analyzer by Technicon - automated instruments have proliferated from many manufacturers: Auto Analyzer (AA) Simultaneous Multiple Analyzer (SMA) series 1970 the first commercial centrifugal analyzer was introduced as a spinoff technology from NASA outerspace research - introduction of the DuPont Automated Clinical Analyzer (ACA) 1975 miniaturization of computers and advancement in the polymer industry for high-grade optical plasctic cuvets 1976 introduction of the thin film analysis technology

1978

production of the Ektachem Analyzer by Kodak Ektachem Analyzer first to use microvolume of sample and reagents on slides for dry chemistry analysis, and it was the first to incorporate computer technology extensively into its design and use 1980 several primary discrete analyzers have been developed that incorporate such characteristics as in ion selective electrodes (ISE), fiber optics, polychromatic analysis, continually more sophisticated computer hardware and software for data handling, and larger test menus Example: Beckman ASTRA, Dade Paramax, Hitachi Analyzers, Chem 1 by Technicon, Abbott Spectrum, and the Olympus and Roche series

1957 Technicon Auto Analyzer

1. High volume of testing 2. Fast turnaround time 3. Expectation of quality results with higher accuracy and precision 4. Intense competition among instrument manufacturers

1. Rapid results 2. Increase in the number of tests performed 3. Saves time and effort 4. Eliminates the need for staff (personnel) increase 5. Economical

6. Errors in calculations and transcription are reduced 7. Better precision and accuracy 8. Uses very small amount of samples and reagents 9. Minimize the variation in results from one laboratorian to another

1. Continuous Flow System 2. Discrete Sampling Analyzers 3. Centrifugal Fast Analyzers 4. Thin Film Analyzers

 Liquids (reagents, diluents, and samples) are pumped through a system of continuous tubing. Smaples are introduced in a sequential manner, following each other through the same network. A series of air bubbles at regular intervals serve as separating and cleaning media.

 Resolves the major consideration of uniformity in performance of tests, because each sample follows the same reaction path Assists the laboratory that needs to run many samples requiring the same procedure This system has won wide acceptance in both routine and research laboratories.

Examples: a. Technicon Autoanalyzer II capable of running 3 different tests at 60-80 samples per hour b. SMA 6/60 capable of running 6 tests at 60 samples per hour c. SMA 12/60 capable of running 12 tests at 60 samples per hour d. SMAC capable of running 40 tests at 120 samples per hour

Simultaneous Multiple Analyzer

Principle: All samples are carried through the same analysis pathway. All samples automatically pass from one step to another without waiting to bring the samples to the same stage of completion. The reactions are not necessarily carried to equilibrium since samples and standards are treated exactly alike.

Features: 1. Use of plastic tubes of different diameters and a peristaltic pump for continuous pumping of samples and reagents. This maneuver replaces the pipetting steps in the manual procedures. 2. Introduction of air bubbles: a. to separate the sample and reagent streams into segments b. to separate one sample from the next c. for continuous scrubbing of tubing d. prevents cross contamination or carry over by the previous specimen

3. Removal of proteins by dialysis. 4. Flow-through cuvettes in an interference filter photometer, using a fixed reference light path. 5. Recorder read-out. 6. Modular design permitting interchanging of major parts.

Parts: 1. Sampler holds the cups containing the standards and specimens for analysis, which are introduced into the analytical system by means of aspiration, in a preset sequence and at a pre-selected rate. 2. Pumps and Manifolds for continuous and proportional delivery of samples, reagents or gases. this is analogous to pipetting in manual techniques.

3. Dialyzer employs dialysis through a semipermeable membrane to separate proteins from the analytes, thus eliminating the need for manual deproteinization techniques. 4. Heating Bath for heating and incubating the reaction mixture at fixed temperatures. 5. Detector-Recorder for continuously measuring and recording optical density or absorbancy changes.

 Separates each sample and accompanying reagents in a separate container Have the capability of running multiple tests one sample at a time or multiple samples one test at a time Most popular and versatile analyzers This system simulates very closely manual procedures except that the various steps are done automatically.

Discrete Sampling Analyzers

Principle: Each sample reactions is handled in a separate compartment and does not come into contact with another sample. The samples and standards are handled on a batch basis and must be brought before proceeding to the next procedure. All reactions must be carried out until equilibrium is reached.

Examples: 1. Dupont ACA (Automatic Cinical Analyzer) Reagents of each test are packaged in a special plastic pack with a rigid header. This pack serves as the reaction chamber and test cuvette for photometric analysis. Each test pack contains: a. chromatographic column that removes interfering substances b. a gel filtration matrix to retard small molecules c. a protein precipitant column 2. Abbott ABA-100 Biochromatic Analyzer, ABA200 and VP Analyzer

Features: a. equipped with a single disposable plastic 32-compartment molding, so that transfer of final solutions for photometry is avoided b. with complete immersion of the reaction vessel in a water bath to achieve rapid rise to stable temperature c. use of ultramicro samples (e.g. 5QL) d. problem of interference from serum and reagent color is overcome by taking absorbance readings at two wavelengths, the socalled bichromatic system

Abbott Analyzer

Hitachi Analyzers

Beckman Analyzers

ASTRA 8 Chemistry Analyzer

Olympus Analyzer

Roche Analyzer

 Uses the force generated by centrifugation to transfer and then contain liquids in separate cuvets for measurement at the perimeter of a spinning rotor Most capable of running multiple samples, one test at a time, in a batch Batch analysis is its major advantage

Centrifuge/ Spinning Rotor

Principle: As the rotor is accelerated, centrifugal force moves the reagents and sample to a mixing chamber and then through a small channel into the cuvette. As the filled cuvette rotates past a fixed light beam, the absorbance of the reaction is measured spectrophotometrically. Examples: 1. CentrifiChem 2. RotoChem

Roto Chem

Principle: A 16 mm square chip which contains several very thin layers, accepts a metered drop of serum, spreads it evenly into a reagent layer, then confines the colored product to a fixed area for reflectance spectrophotometry. Example: 1. Kodak Ektachem

Kodak Ektachem

1. Specimen preparation and identification 2. Specimen measurement and delivery 3. Reagent systems and delivery 4. Chemical reaction phase 5. Measurement phase 6. Signal processing and data handling

1. Specimen Preparation and Identification Specimen / sample preparation generally remains to be a manual process in most laboratories The sample must be properly identified and its location in the analyzer must be monitored throughout the test Labeling a sample cup simplest means of identifying a sample Affixing a bar code label most sophisticated approach in specimen identification

2. Specimen Measurement and Delivery Most instruments use either circular carousels or rectangular racks as specimen containers for holding disposable cups or primary sample tubes in the loading or pipetting zone of the analyzers. these cups or tubes hold standards, controls, and patient specimens to be pipetted into the reaction chambers of the analyzers. The slots in the trays or racks usually are numbered to aid in identification of the sample The trays or racks move automatically in 1-cup step at preselected speeds. The speed determines the number of specimens to be analyzed per hour.

The instrument s computer holds the number cups in memory and aspirates only in slots containing sample cups. The configuration of the sample cup should determine the minimum and maximum values required. Indentations or scorings molded into the plastic can indicate these volumes. The operator can then pour or pipet the samples into the containers easily and quickly. One of the most commonly used containers is the 2mL sample cup.

A sample container

3. Reagents Systems and Delivery Reagents may be classified as: 1. Liquid reagents may be purchased in bulk volume containers or in unit dose packaging 2. Dry reagents packaged in various forms: a. Bottled as lyophilized powder b. Tablet form c. Multilayered dry chemistry slide

Reagent handling techniques used in contemporary analyzers: 1. Keeping all reagents refrigerated until the moment of need and then quickly preincubate them to reaction temperature, or store them in a refrigerated compartment on the analyzer that feeds directly to the disposing area. 2. Providing reagents in a dried, tablet form and reconstitute them when the test is to be run. 3. Manufacturing the reagent in two stable components that will be combined at the moment of reaction.

4. Chemical Reaction Phase This phase consists of: a. Mixing b. Separation c. Incubation d. Reaction Time

In most discrete analyzers, the chemical reactants are held in individual moving containers that are either disposable or reusable. This arrangement allows the analyzer to operate continuously without replacing cuvets. (Examples: Hitachi analyzers and Synchron analyzers)

a. Mixing - a vital component of each procedure - there are different ways of accomplishing mixing depending on the analyzer used: a. Continuous-flow analyzers through the use of coiled tubing b. Centrifugal analyzers centrifugal force is reponsible for the mixing c. ACA analyzers have components specially designed for mixing: the breaker-mixers d. Paramax analyzers uses ultrasonic sound to dissolve the reagent tablet e. Hitachi analyzers use stirring paddles

b. Separation - undesirable constituents that will interfere with an analysis may need to be separated from each sample before the other reagents are introduced into the system (e.g. protein causes major interference in many analyses) - in the older continuous-flow systems, a dialyzer was the separation or filtering module. It performed the equivalent of the manual procedures of precipitation, centrifugation, and filtration.

c. Incubation - Heating bath a component of discrete or continuous-flow systems that maintains the required temperature of the reaction mixture - the principal components of the heating bath are: a. heat-transfer medium (i.e. water or air) b. heating element c. thermoregulator - a thermometer is located in the heating compartment of an analyzer and is monitored by the system s computer

d. Reaction Time - depends on the rate of transport through the system to the read station, timed reagent additions with moving or stationary reaction chambers, or a combination of both processes.

5. Measurement Phase Almost all available systems for measurement have been used, such as ultraviolet, fluorescent, and flame photometry; ion-specific electrodes; gamma counters; and luminometers. Most commonly used system: Visible and ultraviolet light spectrophotometry

Analyzers that measure light require: a. Monochromator used to acheive the desired component wavelength b. Filters/Filter wheels used to separate light c. Fiber optics medium to transport light signals d. Containers holds the reaction mixture

6. Signal Processing and Handling Accurate calibration of instruments is essential to obtaining accurate information because most automated instruments print the results in reportable form After calibration has been performed and the chemical or electrical analysis of the specimen is either in progress or completed, the instrument s computer goes into the data acquisition and calculation mode. All advanced automated instruments have some method of reporting printed results with a link to sample identification. In sophisticated systems, the demographicsample information is entered in the instrument s computer along with the tests required. Then the sample identification is printed with the test results.

 a developed system by the close merging of the 3 phases of the laboratory testing process, that is, pre-analytical (sample processing), analytical (chemical analysis), and postanalytical (data management) phases.