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Hematology Section (Hema Part 1)

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It is a branch of medicine concerning the study of blood, the blood-forming organs and blood diseases.
Hematology
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In the medical field, it includes the treatment of blood disorders and malignancies including types of hemophilia, leukemia, lymphoma and sickle cell anemia
Hematology
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EACMed Hematology Section Offers:
• Complete blood count (CBC)
• Coagulation Studies: Prothrombin time, Activated Partial Thromboplastin Time (APTT) and D-dimer
• Erythrocyte Sedimentation Rate (ESR)
• Reticulocyte Count
• Clotting Time & Bleeding Time
• Malarias Smear
• Peripheral Blood Smear (PBS)
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What specimen is used in Complete Blood Count (CBC)?
EDTA Whole blood (Lavender Top)
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What specimen is used in Erythrocyte Sedimentation Rate (ESR)?
EDTA Whole blood (Lavender Top)
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What specimen is used in Reticulocyte Count?
EDTA Whole blood (Lavender Top)
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What specimen is used in Prothrombin Time?
Citrated Plasma (Blue Top)
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What specimen is used in Activated Partial Thromboplastin Time?
Citrated Plasma (Blue Top)
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What specimen is used in D-dimer?
Citrated Plasma (Blue Top)
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What specimen is used in Clotting Time & Bleeding Time?
Capillary Blood
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What specimen is used in Malarial Smear?
Capillary Blood
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What specimen is used in Peripheral Blood Smear (PBS)?
Capillary Blood
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Turn around time STAT:
1 hour
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It is not applicable for ESR determination
Turn around time STAT: 1 hour
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Hematology Analyzers
• Beckman Coulter Ac.T 5 Diff Analyzer
• Beckman Coulter HmX AL Analyzer
• ACL Elite Pro
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Hematology Analyzer for coagulation tests
ACL Elite Pro
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Hematology Analyzer for CBC test
• Beckman Coulter Ac.T 5 Diff Analyzer
• Beckman Coulter HmX AL Analyzer
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It is a fully automated hematology analyzer providing a complete WBC five-part differential, which is determined simultaneously by the AcV (Absorbance Cytochemistry and Volume) Technology and WBC/
Beckman Coulter Ac.T 5 Diff Analyzer
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Beckman Coulter Ac.T 5 Diff Analyzer Small Sample Volume for CBC:
30 uL
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Beckman Coulter Ac.T 5 Diff Analyzer Small Sample Volume for CBC + Diff:
53 uL
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Beckman Coulter Ac.T 5 Diff Analyzer Throughput
up to 60 samples/hour
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Beckman Coulter Ac.T 5 Diff Analyzer Sample Stability CBC:
48 hours @ room temp.
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Beckman Coulter Ac.T 5 Diff Analyzer Sample Stability Diff:
24 hours @ room temp.
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Beckman Coulter Ac.T 5 Diff Analyzer Reagents
• Diluent, 20 L
• Hgb Lyse, 400 ml
• WBC Lyse
• Diff Fix
• Diff Rinse
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It is used for counting and differentiating blood cells
Diluent, 20 L
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It dilutes whole-blood samples
Diluent, 20 L
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It stabilizes cell membrane for accurate counting and sizing
Diluent, 20 L
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It conducts aperture current
Diluent, 20 L
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It rinses instrument components between analyses
Diluent, 20 L
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It is used to lyse blood cells and to determine hemoglobin concentration
Hgb Lyse 400 ml
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It is used to lyse red blood cells for the leukocyte count and to differentiate poly-nuclear basophils.
WBC lyse
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It is used to lyse erythrocyte, fix leukocytes, and differentially stain granules of monocytes, neutrophils, and eosinophils
Diff Fix
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It is used as a rinsing agent
Diff Rinse
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A hematology quality control material used to monitor the performance of Ac.T 5 Diff analyzers in conjunction with the instrument's reagent system
Ac.T 5 Diff Control Plus
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It is vailable in three levels (low, normal, and high)
Ac.T 5 Diff Control Plus
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Ac.T 5 Diff Control Plus Open stability:
15 days
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Ac.T 5 Diff Control Plus Storage:
2 - 8°C
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Beckman Coulter Ac.T 5 Diff Analyzer Machine Linearity Parameters:
• WBC
• RBC
• Platelet
• Hemoglobin
• Hematocrit
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WBC unit
10^3L
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RBC unit
10^6L
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Platelet unit
g/dL
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Hemoglobin unit
10^3L
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Hematocrit unit
%
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Beckman Coulter Ac.T 5 Diff Analyzer:
WBC Linearity Range
0.5 to 80.0
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Beckman Coulter Ac.T 5 Diff Analyzer:
RBC Linearity Range
0.20 to 7.50
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Beckman Coulter Ac.T 5 Diff Analyzer:
Platelet Linearity Range
10 to 1,000
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Beckman Coulter Ac.T 5 Diff Analyzer:
Hemoglobin Linearity Range
2.5 to 23.0
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Beckman Coulter Ac.T 5 Diff Analyzer:
Hematocrit Linearity Range
11.6 to 55.0
56.0 to 62.4
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WBC Difference (Whichever is Greater)
± 0.2 or ± 3%
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RBC Difference (Whichever is Greater)
± 0.05 or ± 2%
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Platelet Difference (Whichever is Greater)
± 10 or ± 6%
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Hemoglobin Difference (Whichever is Greater)
± 0.3 or ± 2%
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Hematocrit Difference (Whichever is Greater)
± 2 or ± 3%
± 5 or ± 5%
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It is a quantitative, automated hematology analyzer and WBC differential counter for invitro diagnostic use in clinical laboratory
Beckman Coulter HmX AL Analyzer
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Beckman Coulter HmX AL Analyzer Modes of Operation:
• Primary Mode
• Secondary Mode
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Closed-Vial or Automatic Mode
Primary Mode
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Autoloader uses the rocket bed and sensors to move specimen tube holders called cassettes from the loading bay to a cap-baying station, then to the unloading bay
Primary Mode
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Primary Mode aspirates ______ uL of sample from each tube
185 uL
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Open-Vial or Manual Mode
Secondary Mode
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Secondary Mode aspirated ______ uL of sample from each tube via the aspi
125 uL
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Beckman Coulter HmX AL Analyzer Reagents
• ISOTON III diluent
• LYSE III lytic reagent
• Erythrolyse II erythrocyte lytic reagent (also called PAK LYSE)
• StabiLyse leukocyte preservative (also called PAK PRESERVE)
• COULTER CLENZ cleaning agent
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It is also called PAK LYSE
Erythrolyse II erythrocyte lytic reagent
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It is also called as PAK PRESERVE
StabiLyse Leukocyte Preseevative
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The material used for quality control of CBC and DIFF parameters
HmX 5C Cell Control
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HmX 5C Cell Control Three levels of control with varying mixtures of normal and abnormal values of:
Normal
Abnormal 1
Abnormal 2
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HmX 5C Cell Control Storage:
2 - 8°C
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HmX 5C Cell Control Open Stability:
13 days or 13 events ( whichever comes first
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Daily quality control check to verify performance of the VCS technology TTM components in both the DIFF and Retics modes.
Latron Control and Primer
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Contains a surfactant to remove bubbles and clean the sample tubing and pathway from the tip of the aspirate probe to and through the flow cell
Latron Primer
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Contains specific size latex particles with a predetermined volume, conductivity and laser light scatter
Latron Control
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Beckman Coulter HmX AL Analyzer Machine Linearity Parameters:
• WBC
• RBC
• Hemoglobin
• MCV
• Platelet
• MPV
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MCV unit
fL (femtoliters)
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MPV unit
fL
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Beckman Coulter HmX AL Analyzer:
WBC Linearity Range
0.0 - 99.9
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Beckman Coulter HmX AL Analyzer:
RBC Linearity Range
0.0 - 7.00
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Beckman Coulter HmX AL Analyzer:
Hemoglobin Linearity Range
0.0 - 25.0
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Beckman Coulter HmX AL Analyzer:
MCV Linearity Range
50 - 150.0
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Beckman Coulter HmX AL Analyzer:
Platelet Linearity Range
0.0 - 999
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Beckman Coulter HmX AL Analyzer:
MPV Linearity Range
5.0 - 20.0
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It iis an electronic method for counting and sizing particles
Coulter Principle
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it is based on the fact that cells, which are poor conductors of a weak current, will interrupt the current flow.
Coulter Principle
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• The cells are suspended in a conductive diluent and are pulled through the apperture by low vacuum
Coulter Principle
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this creates a momentary increase in resistance to the electric flow.
Coulter Principle
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• The resistance creates a pulse that is sensed and counted by the instrument as a particle
Coulter Principle
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the amount of resistance (the amplitude of each pulse) is directly related to the size (volume) of the particle that produced it
Coulter Principle
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Ac. T 5 Diff Analyzer & HmX AL Analyzer Measurement in WBC
Coulter Principle
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Ac. T 5 Diff Analyzer & HmX AL Analyzer im Hemoglobin
Photometric
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Ac. T 5 Diff Analyzer in Hematocrit
Derived from RBC Histogram
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HmX AL Analyzer Measurement in Hematocrit
Calculated (RBC x MCV) / 10
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Ac. T 5 Diff Analyzer Measurement in Segmenters, Lymphocytes, Monocyte, Eosinophils and Basophils
Impedance with hydrofocus
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HmX AL Analyzer Measurement in Segmenters, Lymphocytes, Monocyte, Eosinophils and Basophils
Derived from WBC scatterplot using VCS technology
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Ac. T 5 Diff Analyzer & HmX AL Analyzer Measurement in Platelet Count
Derived from PIt Histogram
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Ac. T 5 Diff Analyzer & HmX AL Analyzer Measurement in RBC
Coulter Principle
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Ac. T 5 Diff Analyzer Measurement in MCV
Calculated: (Hct/RBC) x 10
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HmX AL Analyzer Measurement in MCV
Derived from RBC Histogram
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Ac. T 5 Diff Analyzer & HmX AL Analyzer Measurement in MCH
Calculated: (Hgb/RBC) x 10
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Ac. T 5 Diff Analyzer & HmX AL Analyzer Measurement in MCHC
Calculated: (Hgb/Hct) x 100
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Ac. T 5 Diff Analyzer & HmX AL Analyzer Measurement in Mean Platelet Volume
Derived form PIt Histogram
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Ac. T 5 Diff Analyzer & HmX AL Analyzer Measurement in Red Cell Distribution Width
Derived from RBC Histogram
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It is a fully automated coagulation system for specific clinical use in hemostasis laboratory for dotting, chromogenic, special assay and immunological testing
ACL ELITE PRO
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What are the tests ACL ELITE PRO used to perform?
• Coagulometric Tests
• Absorbance Tests
• Immunological Tests
• Special Tests
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Coagulometric Tests:
• Protrombin Time
• Activated Partial Thromboplastin Time
• Thrombin Time
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Absorbance Tests:
• Antithrombin
• Protein C
• Plasminogen
• Fibrinogen
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Immunological Tests:
• D-Dimer
• Von Willebrand Factor - Activity and Antigen
•Free Protein S
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Special Tests:
• LAC Screen and Confirm
• Silica Clotting Time Screen and Confirm
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Coagulometric Tests performed in EACMed laboratory
• Protrombin Time
• Activated Partial Thromboplastin Time
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Throughput: Up to _____ samples per hour for PT - Fibrinogen
175 samples
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Throughput: Up to _____ samples per hour for APTT
125 samples
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Turn-around-time: PT/PT - Fibrinogen: ____minutes from standby
4.5 minutes
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Turn-around-time: PT/PT - Fib/APPT: ___ minutes from standby
8 minutes
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ACL ELITE PRO sample tray is _____ samples
40 samples
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ACL ELITE PRO _____ and _____ sample cups
0.5 and 2.0 sample cups
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True or False: ACL ELITE PRO sample tray is a full draw tubes
True
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ACL ELITE PRO reagent positions
Up to 22 total positions
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Ac. T 5 Diff Analyzer Complete Blood Count (Automated): Startup Acceptable Value of WBC
0.3 x 10^3/L
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Ac. T 5 Diff Analyzer Complete Blood Count (Automated): Startup Acceptable Value of RBC
0.03 x 10^6/L
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Ac. T 5 Diff Analyzer Complete Blood Count (Automated): Startup Acceptable Value of Hemoglobin
0.3 g/dL
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Ac. T 5 Diff Analyzer Complete Blood Count (Automated): Startup Acceptable Value of Platelet
7.0 x 10^3L
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After the analysis, review the primer results at the DMS. If count is unacceptable:
Rerun the Latron Primer
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After the analysis, review the primer results at the DMS. If count is acceptable:
Run the Latron Control
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Reconstitute with 8.5 mL PT Buffer (5 days upon reconstitution)
PT Fibrinogen HS Plus
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Liquid ready reagent and has 30 days stability upon opening.
APTT Reagent Synthasil
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Reconstitute with 1 mL distilled water (24 hour stability upon reconstitution).
Normal Control Assayed (Normal Control)
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Reconstitute with 1 mL distilled water (24 hour stability upon reconstitution.
Calibration Plasma (Normal Pool)
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True or False: The leukocytes are counted in the four large squares
True
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True or False: Cells that touch the top and right lines of a square should not be counted.
True
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True or False: Cells on the bottom and left side should be counted.
True
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Review the example computation
Number of leukocytes counted in 4 squares = 188
Number of leukocytes per liter (188 x 0.05) x 10^9
Result Reported: 9.4 x 10^9
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Characteristics of satisfactory smear include the following:
1. There should be no lines extending across or down through the smear.
2. The smear must be smooth at the end, not ragged and lined.
3. The smear must not be too long and too thick.
4. The smear must not contain holes because a greasy slide has been used
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It may be done on a smear to verify a questionable analyzer result.
Platelet estimation
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It is derived by correlating a machine count and a smear count, as follows:
Platelet Estimation Factor
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Platelet Estimation Factor Formula:
MC/SC= iPEF
PEF = Total iPEF ÷ 30

Legend:
MC - Machine Count
SC - Smear Count
iPEF - Individual PEF
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Formula used to calculate the percent erythrocytes
Number of reticulocytes counted/1000 RBC x 1,000 = Reticulocyte ct. in %
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Prothrombin Time Reference Range
11.8 - 15. 2 seconds
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Prothrombin Time % Activity
70 - 100%
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Prothrombin INR
0.80 - 1.20
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Activated partial thromboplastin time (APTT)
25.4 - 38. 4 seconds
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ESR: 0 - 18 years
0 - 10 mm/hr
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ESR: > 18 years (Male)
0 - 15 mm/hr
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ESR: > 18 years (Female)
0 - 20 mm/hr
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Reticulocyte Count: > 18 years (Male)
0.5 - 1.5 %
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Bleeding Time: > 18 years (Male)
1 - 3 minutes
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Clotting Time: > 18 years (Male)
3 - 5 minutes
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It aims to help laboratories attain accurate and reliable analytical results and achieve inter-laboratory comparability as well.
National External Quality Assessment Scheme (NEQAS) Enrollment
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It is one of the requirements for the renewal of the License to Operate (LTO) of Clinical Laboratories of all category.
National External Quality Assessment Scheme (NEQAS) Enrollment
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Reference Laboratory for Hematology:
National External Quality Assessment Scheme (NEQAS) Enrollment
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