details about hematology automation

1. GUIDELINE ABOUT
COURSE
Hematology test and techniques
MotherTarrasaVocationalTraining Institute
Present by-
Dikshit Prajapati

2. Objects
■ Introduction of Hematology
■ Hematology test
■ Instruments andAutomation

3. Introduction to Hematology
■ Hematology is the study of blood and blood disorders. Hematologists and
hematopathologists are highly trained healthcare providers who specialize in diseases
of the blood and blood components.These include blood and bone marrow cells.
Hematological tests can help diagnose anemia, infection, hemophilia, blood-clotting
disorders, and leukemia.

4. Common hematology tests
Test Uses
Complete blood count (CBC), which includes:
• White blood cell count (WBC)
• Red blood cell count (RBC)
• Platelet count
• Hematocrit red blood cell volume (HCT)
• Hemoglobin concentration (HB).This is the
oxygen-carrying protein in red blood cells.
• Differential white blood count
• Red blood cell indices (measurements)
To aid in diagnosing anemia, certain cancers of the
blood, inflammatory diseases, and to monitor blood
loss and infection
Platelet count (usually done as part of the
CBC)
To diagnose and/or to monitor certain types of
bleeding and clotting disorders
Prothrombin time (PT) Partial
Thromboplastin Time (PTT) International
To evaluate bleeding and clotting disorders and
to monitor anticoagulation (anticlotting)
therapies
Bone marrow biopsy is not a common test in general, but is a common test for hematologists. It involves
taking cells from the bone marrow for analysis for many types of disease.

5. Some OtherTest
■ Full blood count
■ Blood film
■ Staining may also detect blood parasites such as malaria, toxoplasmosis, and
microfiliariasis.
■ Assessment and staining of immature platelets or megakaryocytes may also be
performed.
■ erythrocyte sedimentation rate (ESR)
■ The bone marrow may be examined.
■ Iron status and anemias
■ Platelet function in bleeding and coagulation
■ The enzyme G6PD may be assessed in sickle cell disease.
■ Etc,.

6. Instruments & Automatio in CBC
■ A complete blood count (CBC) is a series of tests used to evaluate the composition and
concentration of the cellular components of blood.
It consists of the following tests:
1. Red blood cell (RBC) count,
2 .White blood cell (WBC) count, & differentiation.
3 . Platelet count (Plts),
4 . Measurement of hemoglobin (HgB) &
hematocrit (HCT or PCV)
5 . Red blood cell indices (MCV, MCH,MCHC,RDW)

7. Method for CBC
1. Manual methods
2. Automation
1. Semi-Automation
2. Fully Automation

8. Manual Methods for CBC
■ Haemoglobinometry method – Hb
■ Heamoglobinocytrometry method – RBC,WBC, PLT
■ Staining slide Method – DC forWBC , premature RBC count, Retic Count,Red Cell
Indices
■ Wintrob &Westergen Method – PCV & ESR
■ Dukes Method – BT
■ Wright’s or Capillary tube method – CT
■ Etc.,

9. Automation in CBC
■ Cell Counter - Hematology analyzers are computerized, highly specialized and
automated machines that count the number of different kinds of white and red blood
cells in a blood sample.

10. Types of Counters
Semi automated :
• Some steps carried out manually like
dilution of blood Measures only a few
parameters
• Semi-automated instruments
Require some steps, as dilution of
blood samples.Often measure only a
small number of variables.
Fully automated:
• Require only anticoagulated blood
samples. Measures multiple
parameters
• Fully automated instruments Require
only that an appropriate blood
sample is presented to the
instrument.
• They can measure 8-20 variables
including some new parameters
which do not have any equivalent in
manual methods.

11. Advantages of electronic cell counter
■ Rapid & shortens the time of tests and complete a large
number of tests quickly
■ Reduce the cost of the tests
■ Reduce the testing equipments (instruments)
■ Maintain and improve accuracy of tests

12. 18 Parameter electronic cell counter
1. WBC: White Blood Cells
2. RBC: Red Blood Cells
3. HgB: Hemoglobin
4. Hct: Hematocrit
5. MCV: Mean Cell Volume
6. MCH: Mean Cell Hemoglobin
7. MCHC: Mean Cell Hemoglobin Concentration
8. RDW: Red Blood Cell Distribution Width

13. 18 Parameter Cell Counter
8 parameters plus
9. LYM#: Lymphocytes total
count
10. MID#: Mid Cells total
count
11. GRN#: Granulocytes total
count
12. LYM%: Lymphocytes
Percentage
13. MID%: Mid Cells
Percentage
14. GRN%: Granulocytes
Percentage
15. Pct: Plateletcrit
16. PDW: Platelet
Distribution Width
17. WBC histogram
18. Plt Histogram

14. Electronic blood cell counter
■ Two General Principles
1. Electronic resistance ( impedance)
2. Light scattering
other –
3. Use of Fluorescent Dyes
4. Flowcytometry
5. Radiofrequency
■ Counts blood cells by electronic impedance (resistance).
■ Discrimination & differentiation of blood cells based on cell size.
■ Estimation of hemoglobin by Cyanomethemoglobin method using optical way.
■ Reagents and diluent solution included.

16. The Coulter (Impedance) Principle of hematology analyzer
■ Aperture... A small opening (orifice) in a
device by which blood cells may be drawn
into an analyzer for counting.
■ Principle. Electrical impedance.. Sizing
and counting cells by detecting and
measuring changes in electrical resistance
when a cell passes through a small aperture.
■ Tens of thousands of cells are counted,
providing low statistical deviations with an
accuracy of measurement estimated by
Coulter to be within 1% of the true count

17. ■ A small sample of the well mixed blood is aspirated and diluted with
a balanced isotonic saline solution.
■ The diluted blood sample is split into two parts, one for counting
RBCs and platelets and the other for counting WBCs.
■ The portion of the WBC bath and a detergent (lysing agent) is
added to destroy (hemolyze) the red blood cells.
■ The lysing agent contains potassium cyanide that reacts with the
hemoglobin to form cyanmethemoglobin & the hemoglobin
concentration is measured optically.
■ The results are printed out or sent to a computer for review.

19. Sysmex xs 1000i

20. Sysmex Instrumentation
How data are reported
Principles of Measurement
Direct Measurement:
RBC – (Impedance),
WBC – (Impedance),
Platelets – (Impedance), (2-30 fl)
Hgb – SLS-Hb (555 nm) ( sodium lauryl
sulfate)
HCT – cumulative pulse height detection
Indirect Measurement:
MCV, MCHC, and MCH (calculations)
RDW and MPV (CV of respective histograms)
WBC differential
Impedance ( CD)/RF detection
employs differential lysis
Flags

21. WBC differential plot generated by all analyzers shows a
similar cell distribution pattern

23. Red Cell Histogram
■ Represents the relationship between RBCs size and number
Normal red cell histogram displays cells form (36- 360 ) fl
If RBCs are larger than normal shift to the right
If RBCs are smaller than normal shift to the left
If the curve is bimodal 2 population of RBCs

24. Platelets Histogram
The share of platelets >12 fL in the
total platelet number is presented in %.
The standard range is 15–35%. An
increase of the parameter may be an
indication for :
platelet aggregates, microerythrocytes
and giant platelets.

25. WBC Histogram

27. When to Calibrate
You should calibrate your instrument:
■ At installation
■ After the replacement of any component that involves
dilution characteristics or the primary measurements
(such as the apertures)
■ As a routine once or twice a year

28. (Daily Maintenance)
■ Turn on the analyzer. On initial start up, the instrument will perform self-
checks
■ If all the self-checks is satisfactory, then the instrument is ready for
analysis.
■ If an error message is displayed on the analyzer screen , an alarm will
sound.
The following corrective action will need to be taken before analysis can
go ahead:
■ Silence the alarm.
■ Press Help to display the error message
■ Press OK and the relevant corrective action will be automatically
performed
■ Run the internal QC after start up procedure is completed.

29. Weekly maintenance
■ Check the quality control chart for evidence of drift.
■ Check the daily averages of (MCV, MCH and MCHC) for any drift or
sudden change.
■ Clean the orifice and cell with a fine brush and flush several times
with diluents. Never attempt to clear the orifice with a sharp device
such as a needle or blade.
■ Check seals to determine the possibility of leakage.
■ Check tubing.
■ Check stock of reagents, diluents, and disposables.
■ In a special logbook record the dates of all maintenance checks,
replacements of components, servicing by manufacturer's agent,
recalibrations, and other necessary information.
■

30. Problem Solving –Troubleshooting
■ An instrument problem is differentiated from a
specimen-related problem by running a control.
■ If the control results are acceptable, the problem is
probably specimen-related. Check for:
– clots
– hemolysis
– lipemia

31. Some cell counter with fetures
ABX Micros ES 60
■ Highly compact and lightweight
■ Micro-sampling on whole blood: 10μL
■ Precise cellular identification through
electronic impedance variation method
■ Haemoglobin measurement with
photometry
■ Integrated barcode reader (samples, QC
and reagents) and thermal printer (full data
printout and graphs)
■ Embedded Data Management: Data
transfer with USB key to archive or
download data, high results capacity,
customized reports.
■ Compatible with innovative reagent pack
or bottle systems
• 60 samples per hour (OT
Version)
• CBC + 3 DIFF (18 parameters)
• Large color touch screen

32. Mindray BC-3200
■ 3-part differentiation ofWBC ;
19parameters + 3histograms
■ Closed tube sampling
■ Sampling with 4 position tube holder
for various tubes
■ Micro sampling of 13 μL for whole
blood
■ 2 counting mode: whole blood and
prediluted
■ Up to 60 samples per hour
■ Large color LCD display
■ Data storage of 35 , 000 samples
with histogram

33. Mindray Pentra XLR
Compact and reliable hematology platform including reticulocytes by fluorescence
■ 6 parameters, 10 reticulocyte
parameters
■ Up to 80 samples / hour
■ Enhance productivity, quality,
flexibility
■ Increase your expertise in anaemia
characterization
■ Add value to your hematology lab
■ Throughput: Up to 80 samples/hour
■ Large capacity auto-loader (100
tubes
■ High resolution reticulocyte Matrix
for three stages of reticulocytes
maturation quantification

34. Other Cell counter name
■ Medonic M32B
■ Medonic M20
■ Adonis Axiom 19
■ HCT Mindray
■ Sysmex xp 100
■ Horiba H500
■ Etc,.

36. Questions?