Mohd Hairi Bin A.Hamid Bsc.

Medical Laboratory Technology (Hons) UniversitI Teknologi Mara, Malaysia (July 2009)

Automation in haematology lab

Automated cell counted (Full blood count test) Advantages of automation 1) High capital 2) Rapid performance 3) Less labour 4) More precise 6) No errors 7) Improved quality of test 8) Reduce cost of the running test 8) Need calibration and maintenance

Hb
Red cell (Quantitative measurement)

PCV/ Hct

RBC indices
Reticulocytes

RBC Count

Hb count (Automation) Direct measurement Modification of manual HiCN method (Cyanmethemoglobin method) o Concentration of reagent, temperature and pH of reaction o Addition of non-ionic detergent to ensure rapid cell lysis, reduce turbidity o Measurement at set interval before the reaction is completed o Utilization of non cyanide reagent Another technique: High-angle light scattering method
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Laboratory Automation

Mohd Hairi Bin A.Hamid Bsc. Medical Laboratory Technology (Hons) UniversitI Teknologi Mara, Malaysia (July 2009)

PCV/ Hct (Automation) Passage of a cell through the aperture of an impedance counter or through the beam of light in a light scattering instrument Lead to the generation of an electrical impulse (Direct obtained) Another technique: (MCV X RBC) / 10

RBC count (Automation) Electrical impedance

Example: Beckman-Coulter, Sysmex, Abbott, Roche RBCs are poor conductors of electricity and therefore it was diluted in a buffered electrolyte solution Passage of a cell via the aperture of an impedance counter lead to the generation of an electrical impulse All electrical-impedance cell counters are based on Coulters principle.

No of pulses = count as cell Average pulse height = volume Summation of height = PCV

Cells through an aperture, causes a change in electrical resistance, this pulse is detected and amplified by the instrument. Amplitude of the pulse is proportional to cell size
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Laboratory Automation

Mohd Hairi Bin A.Hamid Bsc. Medical Laboratory Technology (Hons) UniversitI Teknologi Mara, Malaysia (July 2009)

Light-scattering technology (cell counting) Diluted cell suspension flow in a single file A light source in front of the aperture (strike a cell; the beam is scattered at an angle). The angle is depend on the volume, size and index of the refraction Scatter lights is detected by photomultiplier or photodiode And converted into the electrical impulses (the values are read at readout device)

Red cell indices (Automation) Red cell distribution width (HDW) Technique: Calculation o RDW CV = SD X 100 MCV o RDW SD = SD of RBC histogram

Laboratory Automation

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Mohd Hairi Bin A.Hamid Bsc. Medical Laboratory Technology (Hons) UniversitI Teknologi Mara, Malaysia (July 2009)

Hemoglobin distribution width (HDW) SD of Hb concentration distribution histogram Only obtained from Laser technology Degree of anisocytosis can be determined It derived from pulse height analysis CV (%) or SD (fl)

Mean Corpuscular haemoglobin concentration (MCHC) Technique: Calculation o Hb X 100 Hct Mean Corpuscular haemoglobin (MCHC) Technique: Calculation o Hb X 10 Hct Corpuscular haemoglobin concentration means (CHCM) Technique: Direct measurement using light scattering at different angle (5 15C) in Bayer-Technicon To replaced the role of MCHC. Sensitivity to iron deficiency has improved. As an internal quality control. If all measurement accurate, MCHC = CHCM

Reticulocytes count (Automation) Technique: o Fluorescence detection of red cells stained with RNA specific fluorochroms ( Auramine O, Ethidium bromide, Oxazine 750, Thiazole orange) o Direct count via volume, light scattering and opacity of cells Type after fluorescence detection: o Low Fluorescence Ratio (LFR): Most mature o Middle Fluorescence Ratio (MFR): mid mature o High Fluorescence Ratio (HFR): Most immature

Laboratory Automation

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Mohd Hairi Bin A.Hamid Bsc. Medical Laboratory Technology (Hons) UniversitI Teknologi Mara, Malaysia (July 2009)

Total WBC count (Automation) Electrical Impedance (Number of pulse obstructed) @ Light scattering (Number of signal from low-angle light scattering) WBC count: o Red cells are lysed, residual particles are counted o Threshold are set for WBC to exclude platelet o Error for WBC: giant platelet, nRBC, white cell agglutination

Platelet count (Automation) Electrical Impedance (Volume cut-off at 2fl-20fl) @ Light scattering (High-angle light scattering for size and Hb content) Platelet count: o Counted in WB using electrical electro-optical detection o Threshold is set to separate red cells, debris and electronic noise o New plates parameter: MPV, PDW

WBC Differential count (Automation) More precise but sometimes inaccurate Using flow cytometry principle Diluted WB, RC lysed, WC categorised into 3 or 5 part diff Single channel or 2/ more channels Based on: Volume, Physical characteristics, Activity of cellular enzyme Techniques: o By volume o By volume + conductivity + light scattering o By light scattering + cytochemistry o By light scattering + RF impedance 3 part Single channel Based on volume of various cells 3 categories detected: o Granulocytes/LC o Lymphocytes/SC o Monocytes/MNC Eosinophil/basophiles are included in MNC 5 part or more 2 or more channel Cell volume and characteristics 5 categories detected: o Neutrophils o Eosinophils o Basophils o Lymphocytes o Monocytes Other may have LIC, AL...

other

Laboratory Automation

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Mohd Hairi Bin A.Hamid Bsc. Medical Laboratory Technology (Hons) UniversitI Teknologi Mara, Malaysia (July 2009)

Instrument principle Light scattering and absorbance Impedance measurement with low and high frequency electromagnetic current or radiofrequency current Cytochemical reaction

Analysis 2 parameter or more complex Cells are divided into cluster Threshold (fixed and variable)

Laboratory Automation

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Mohd Hairi Bin A.Hamid Bsc. Medical Laboratory Technology (Hons) UniversitI Teknologi Mara, Malaysia (July 2009)

Overall result may obtained when running the test

Laboratory Automation

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Mohd Hairi Bin A.Hamid Bsc. Medical Laboratory Technology (Hons) UniversitI Teknologi Mara, Malaysia (July 2009)

References 1. Robin A. Felder (1999), Laboratory Automation: State of the Art and Trends.pdf, Medical Automation Research Centre, Charlottesville, VA retrieved on September 27, 2009 from www.scribd.com 2. Prof. Dr. Thanusak Tatu (2002) Automated Blood Cell Analyzer.ppt, Chiang Mai University retrieved on September 27, 2009 from www.scribd.com 3. Dr Eow Geok Im (2004), Basic Technique in Haematology.pdf, Malaya University retrieved on September 27, 2009 from www.scribd.com 4. Automated analyzer (September 12, 2009) retrieved on September 28. 2009 from http://en.wikipedia.org/wiki/Automated_analyser 5. Laboratory automation (July 26, 2009) retrieved on September 28, 2009 from http://en.wikipedia.org/wiki/Laboratory_automation 6. Complete blood count (September 27, 2009) retrieved on September 28, 2009 from http://en.wikipedia.org/wiki/Complete_blood_count 7. Automation in haematology (2007) retrieved on September 28, 2009 from http://www.bloodindex.com/view_workshop_master.php?id=36

Laboratory Automation

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