Chapter 1 : Clinical Laboratory Instrumentation

Introduction Provide Information to : Aid in the diagnosis of disease

Patient specimens

Analysis by Clinical Laboratory

Evaluate the effectiveness of therapy

Department of clinical laboratory = department of clinical pathology = department of laboratory medicine

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Chapter 1 : Clinical Laboratory Instrumentation

Sections of Clinical laboratory

Chemistry : blood , urine, other fluids to determine how much of

various clinically important substances they contain. (Most applications of electronic instrumentation) Hematology : determines of the numbers and characteristics of the formed elements in the blood (red cells, white cells, platelets) also tests the function of physiological systems in the blood (clotting) Microbiology : performs studies on various body tissues and fluids to determine whether pathological microorganisms are present Blood bank

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Many critical patient care decisions are based on tests results supplied by the clinical laboratory, the accuracy and precision of these results are of great importance. Excellent equipment design and effectiveness quality control programs are essential. Erroneous results can lead to tragic outcome. We should have a fast response in many test procedures : the application of electronic instrumentation has greatly reduced the time required to perform these tests Laboratory information systems : Keep track of specimens Organize the flow of work Automatically acquire test results from some types of instruments Maintain test results databases Report results online
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Spectrophotometry Its a technique that is considered to be the basis for many of the instruments used in clinical laboratory due to the ease of measurement, satisfactory accuracy and precision, and the suitability of this technique to use in automated systems. Spectrophotometry is based on the fact that substances of clinical interest selectively absorb or emit electromagnetic energy at different wavelengths. From ultraviolet (200 400 nm) , visible (400 700 nm) to the near infra red (700 800 nm) Majority of instruments operate in the visible range.

X-Ray 200nm

UV 400nm

Visible 800nm

IR

Microwave

WAVELENGTH(nm)
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General block diagram of Spectrophotometry type instrument

The light source supplies the radiant energy used to analyze the sample. The wavelength selector allows energy in a limited wavelength band to pass through The sample will be put in a cuvette. The detector produces an electric output proportional to the amount of energy it receives, the meter may indicate the received energy or some function of it (such as the concentration of a substance)
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General block diagram of Spectrophotometry type instrument

The main principle of this technique is that if we examine an appropriately chosen sufficiently small portion of the electromagnetic spectrum, we can use the energy absorption properties of a substance of interest to measure the concentration of that substance. In most cases these substances do not exhibit the desired energy absorption characteristics, reagents must be added to the sample in order to cause a reaction to occur.

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General block diagram of Spectrophotometry type instrument Light sources Hydrogen and deuterium discharge lamps are used to provide power in the 200 to 360 nm range. They produce continuous spectrum, the problem is that they produce about 90 % of their power in the infrared range. The ultraviolet and visible range can be increased by increasing the value of voltage. Tungsten filament lamps are used for 360 to 800 nm During operation , the lamp vaporizes from the filaments and condenses on the glass envelope, the coating used may cause errors in determination

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General block diagram of Spectrophotometry type instrument Wavelength selectors These devices used to select portions of the power spectrum that are to be used to analyze the sample. They are divided into two classes Filters Glass filter Interference filter Monochromators

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Glass filters function by absorbing power, for example a blue colored filter absorbs the red region and transmits the blue green region. They are designed to be low-pass, highpass and band-pass filters (low accuracy) Interference filters are made by spacing reflecting surfaces such that the incident light is reflected back and forth for a short distance. The distance is selected such that light in the wavelength band of interest to be in phase and to be reinforced, light outside the band is out of phase and canceled. (used in many spectrophotometers) Monochromators are devices that utilize prisms and diffraction gratings. They provide narrow bandwidths and have adjustable nominal wavelengths. The basic principle is that they disperse the input beam spatially as a function of wavelength. A mechanical device is then used to allow wavelengths in the band of interest to pass through a slit.

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Chapter 1 : Clinical Laboratory Instrumentation Prisms are constructed from glass and quartz
Infrared

monochromatic Ray
Red Orange Yellow Green Blue Violet

SLIT

Polychromatic Ray

PRISM

Ultraviolet

Polychromatic Ray

Monochromatic Ray

A convergent lens system is used to direct the light from the source through an entrance slit. The nonlinear spatial distribution of the power emerging from a prism requires complex mechanical devices for control of the slit position to select different nominal wavelengths.
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Diffraction gratings are constructed by inscribing a large number of closely spaced parallel lines on glass or metal. A grating exploits the fact that rays of light bend around sharp corners, the degree of bending is a function of wavelength

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Chapter 1 : Clinical Laboratory Instrumentation General block diagram of Spectrophotometry type instrument Cuvettes

The cuvette holds the substance being analyzed. Its optical characteristics must be such that it does not alter the spectral characteristics of the light as that light enters or leaves the cuvette. The cuvettes come in a variety of shapes and are constructed of several different materials. The faces (part in the light path) may be silica, quartz, glass, or plastic, but must be transparent to the radiation of interest.
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Chapter 1 : Clinical Laboratory Instrumentation General block diagram of Spectrophotometry type instrument Samples
The sample (patient specimen + reagent) absorbs light selectively according to Beers Law Equal thickness of an absorbing material will absorb a constant fraction of the energy incident upon it
P = P010 aLC P0 radiant power arriving to the cuvette P radiant power leaving the cuvette a absorptivity of the sample L Length of the path trough the sample C Concentration of the absorbing substance P %T = 100 = 100(10 aLC ) P0 This is the percent transmitance a, L are constants, so changes in P will reflect changes in the concentration of the absorbing substance in the sample
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P A = log 0 P

where A = aLC (Absorbance)

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Chapter 1 : Clinical Laboratory Instrumentation General block diagram of Spectrophotometry type instrument Samples
P watt/m 2 S The concentration of an unknown can be determined as follows, the absorbance As of a standard with known concentration of the substance of interest Cs is determined. I=

The concentration of the unknown Cu is computed via the relationship

Cu = Cs

Au As

For multiple values of Cs, we measure As then we plot the line of correlation

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Chapter 1 : Clinical Laboratory Instrumentation General block diagram of Spectrophotometry type instrument Samples

2 methods of measurement : final point : we put the sample with a reagent, we wait until the reaction is done, then we measure the concentration enzymatic reaction : we put the sample with a reagent, the reaction starts but still in progress, we measure the concentration for different values of time. For multiple values of Cs, we measure As then we plot the line of correlation
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Chapter 1 : Clinical Laboratory Instrumentation General block diagram of Spectrophotometry type instrument Photometric system

A spectrophotometers photometric system includes detectors to measure the amount of power leaving the cuvette, circuits for amplification of the low currents developed by detectors, and devices to present the results of the determination to the technologist operating the instrument. Commonly used detectors include barrier layer cells, phototubes, and photoconductive cells.

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General block diagram of Spectrophotometry type instrument Photometer 1 beam, 1 cell

Photometer 1 beam, 2 cells

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General block diagram of Spectrophotometry type instrument Photometer 2 beam, 1 cell

The standard is a solution with known concentration This system is used to perform a continuous calibration We use a unique standard and we make a comparison at the output
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Au Cu = Cs As
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