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 6) No errors

2) Rapid performance 7) Improved quality of test
3) Less labour 8) Reduce cost of the running test
4) More precise 8) Need calibration and maintenance

Hb PCV/
Red cell Hct
(Quantitative
measurement)
RBC RBC
indices Count
Reticulocytes

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

Laboratory Automation Page | 1

 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
Laboratory Automation Page | 2
 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 Page | 3

 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 Page | 4

 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 5 part or more

Single channel 2 or more channel
Based on volume of various cells Cell volume and other
3 categories detected: characteristics
o Granulocytes/LC 5 categories detected:
o Lymphocytes/SC o Neutrophils
o Monocytes/MNC o Eosinophils
Eosinophil/basophiles are included in o Basophils
MNC o Lymphocytes
o Monocytes
Other may have LIC, AL...

Laboratory Automation Page | 5

 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 Page | 6

 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 Page | 7

 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 Page | 8