AUTOMATED CELL COUNTING INSTRUMENTATION & POINT OF CARE TESTING

Advantages
Provides greater accuracy and precision than manual methods Multiple tests can be done instantly Can provide 8 standard hematology parameters(CBC plus RBC indices) plus differential count in less than 1 minute Requires small amount of anticoagulated blood

General Principles of Hematology Analyzers

Electronic impedance: low voltage direct current resistance, most commonly used Radiofrequency: alternating current resistance; a modification sometimes used in conjunction with DC electronic impedance Optical scatter: uses both laser and nonlaser light

ELECTRONIC IMPEDANCE
Based on the detection and measurement of changes in electrical resistance produced by cells as they traverse a small aperture

ELECTRONIC IMPEDANCE
1. 2. 3. 4. diluent: eg saline Aperture: orifice Electrodes (external and internal) Oscilloscope screen : display the pulses generated as cell passes thru the aperture *# of pulses: directly proportional to # of cells Size of voltage pulse: directly proportional to size (volume) of the cell

RADIOFREQUENCY (RF)
High voltage electromagnetic current flowing between both electrodes simultaneously Can measure the cell interior density (eg nuclear volume) Can measure conductivity of the cells (attenuated by the nuclear-to-cytoplasmic ratio, nuclear density, and cytoplasmic granules)

OPTICAL SCATTER
Uses flow cytometry (hydrodynamically focused sample stream is directed thru quartz flow cell past a focused light source) Light source is a tungsten-halogen lamp or helium-neon laser Monochromatic light is emitted (light w/ single wavelength)

OPTICAL SCATTER
As cell passes thru the sensing zone and interrupt the light, light is scattered in all directions Forward angle light scatter (0 degree) correlates with cell volume or size Orthogonal light scatter (90 degrees) or side scatter correlates with degree of internal complexity

OPTICAL SCATTER
Forward low-angle scatter (2-3 degrees) and forward high-angle scatter(5-15 degrees) correlate with cell volume and refractive index or with internal complexity.

PRINCIPAL CELL COUNTING INSTRUMENTS

Beckman Coulter, Inc. : uses impedance Sysmex Corporation : uses impedance and RF Abott Laboratories : impedance and optical measurements Bayer Corporation: optical flow cytometry

PRINCIPAL CELL COUNTING INSTRUMENTS

Common parts 1. Hydraulics: includes aspirating units, dispensers, diluters, mixing chambers, aperture baths, hemoglobinometer 2. Pneumatics: vacuums and pressures 3. Electrical system: electronic analyzers, computing circuitry for data processing

AUTOMATED RETICULOCYTE COUNTING

Retic Count: last to became automated May count 32000 RBCs (1000 RBCs for manual) to improve accuracy Uses fluorescent or nucleic acid dye

LIMITATIONS AND INTERFERENCES

Use internal QC (calibrators) : recommended by CLSI Calibration is crucial in defining accuracy. Calibration is the process of electronically correcting an instrument for analytical bias (numerical difference from the true value) Calibration is accomplished by using reference materials, reference methods, and calibrators

LIMITATIONS AND INTERFERENCES

Accdg to CLSI, calibration must be performed at initial installation of the analyzer and verified at least every 6 months. Limitations of the machine is classified as instrument limitations and sample limitations Instrument limitation is related to methodology. A common method limitation is an instruments inability to distinguish cells. Flagging of machine indicates possible rejection of automated results.

LIMITATIONS AND INTERFERENCES

Sample limitations include factors such as cold agglutinins, icterus, and lipemia Sample age and improper sample handling can affect reliability of test results.

POINT OF CARE (POC) TESTING

A diagnostic lab test at or near the site of patient care Testing site neutrality: t does not matter where the diagnostic test is performed and who performed the test CLIA TESTING is classified as : waived, moderately complex, highly complex

 Waived tests: simple with an insignificant risk of erroneous result Examples of waived test are glucose, Hgb, Hct, Pregnancy and urine testing strip Non waived tests are activated clotting time, prothrombin time, activated partial thromboplastin time)

POC ADVANTAGES and DISADVANTAGES

Fast results : advantage Disadvantages: 1. More expensive 2. Usage 3. Oversight