FLOWCYTOMETRY

FLOWCYTOMETRY

FLOWCYTOMETRY
B5073 Immunophenotyping of Leukaemia BY FLOWCYTOMETRY

Selective panels of antibodies are used for cell surface/intracellular markers to evaluate and diagnose specific Acute Leukaemia types based on the clinical diagnosis and other primary lab findings. Report interpreted based on the percentage of expression of CD markers.


Markers used for AML, ALL (T-ALL, Pre-B-ALL): Cell surface:

CD56, CD13, CD34, CD19, CD10, CD38, CD20, CD64, CD33,
CD11c, CD117, CD11b, CD14, HLADR.

Cytoplasmic: MPO, cCD79a, cCD3,

Secondary Markers for AML –M6 and AML-M7

CD61 FITC, CD41 PE, CD45 CP, CD235a

Secondary Markers for T-ALL

CD4, CD1a, CD5, CD10, CD2, CD3, CD8

Gating Strategy: CD45/SSC Gating.

5-6 ml of Sodium heparin (Green top) anti-coagulated peripheral blood, bone marrow samples.

2-3 ml of other body fluids pericardial fluid, ascitic fluid, CSF, pleural fluid.

Storage: Bone marrow samples in room temperature.

Peripheral Blood can be stored at 4°C for processing next day.

CSF must be freshly processed.  
B5080 Minimal Residual Disease detection (MRD) for B cell ALL

Selective panels of antibodies are used for cell surface markers to evaluate minimal residual disease based on the clinical diagnosis and other primary lab findings.


Markers Used for MRD of B CELL ALL:

CD20, CD10, CD19, CD22, CD34, CD58, CD11a, CD38 , CD123

Gating Strategy: CD45/SSC, CD19/SSC Gating.


5-6 ml of Sodium heparin (Green top) anti-coagulated bone marrow samples.  
B5076 Immunophenotyping of Lymphoma BY FLOWCYTOMETRY

Selective panels of antibodies are used for cell surface markers to evaluate and diagnose specific Lymphomas based on the clinical diagnosis and other primary lab findings.

Markers used for Lymphoma diagnosis (T cell, B-cell):

CD23, CD10, CD79b, CD19, CD200, CD43, CD20 LAMBDA, KAPPA, CD5, CD19, CD3, CD38, CD7

Secondary Markers for BURKITT'S LYMPHOMA (ALL-L3): Lambda, Kappa, CD20


Secondary Markers for B-NHL: CD25, CD103, CD11c, CD19, CD123, CD22, CD45, IgM, IgD, CD19


Secondary Markers for T-NHL:CD57, CD16, CD3, CD56, CD25, CD8, CD4, CD45, TCRαβ, TCRγδ, CD3, CD2


Gating Strategy: CD45/SSC, CD19/SSC Gating.

5-6 ml of Sodium heparin (Green top) anti-coagulated bone marrow samples.
B5073 Immunophenotyping of Wiscott Aldrich Syndrome (WAS) BY FLOWCYTOMETRY

Markers Used for WAS diagnosis:

WASP B9 and WASP D1 on healthy control, mother and patient.

Gating strategy: FSC/SSC on lymphocytes

5-6 ml of Sodium heparin anti-coagulated (Green top) peripheral blood samples of patient, patient's mother and a healthy control.
B5074 Immunophenotyping OF ALPS (double negative T cells) BY FLOWCYTOMETRY

Selective panels of antibodies are used for cell surface markers to evaluate and diagnose ALPS based on the clinical diagnosis and other primary lab findings.


Markers used for ALPS Diagnosis:

CD3, TCRαβ, CD4, CD8, CD56, CD20 on healthy control and patient.

Gating strategy: FSC/SSC for total lymphocytes or CD3/SSC plot for gated T lymphocytes

5-6 ml of Sodium Heparin anti-coagulated (Green top) peripheral blood samples of patient and a healthy control.
B5074 IMMUNOPHENOTYPING FOR LAD (Leukocyte Adhesion Deficiency) BY FLOWCYTOMETRY

Selective panels of antibodies are used for cell surface markers to evaluate and diagnose LAD based on the clinical diagnosis and other primary lab findings.

Markers used for LAD Diagnosis:

CD18, CD11a, CD11b, CD11c on patient and healthy control

Gating strategy: FSC/SSC on granulocytes

5-6 ml of Sodium Heparin anti-coagulated (Green top) peripheral blood samples of patient and a healthy control.
B5074 Immunophenotyping of severe combined immunodeficiency disorder (SCID) / LYMPHOCYTE SUBSET ANALYSIS BY FLOWCYTOMETRY

 Selective panels of antibodies are used for cell surface markers to evaluate and diagnose SCID / Analyze Lymphocyte subsets based on the clinical diagnosis and other primary lab findings. Report interpreted as absolute counts based on the percentage of expression of CD markers.


Markers used for SCID / Subset Analysis:

CD4, CD8, CD 3, CD19, CD56, CD45

Gating Strategy: CD45/SSC on lymphocytes.

5-6 ml of Sodium heparin anti-coagulated (Green top) peripheral blood sample of patient. Stored at 4°C.
B5074 Platelet Immunophenotyping for Glanzmann’s Thrombasthenia (GT) BY FLOWCYTOMETRY

 Selective panels of antibodies are used for cell surface markers to evaluate and diagnose GT based on the clinical diagnosis and other primary lab findings.


Markers used for GT Diagnosis:

CD42a (GPIb /IX) and CD41a (GPIIb) and CD61 (GPIIIa) on healthy control and patient.

Gating strategy: FSC/SSC on platelets

5-6 ml of Sodium heparin anti-coagulated (Green top) peripheral blood samples of patient and a healthy control.
B5075 Flow cytometry for Paroxysmal Nocturnal Hemoglobinuria (PNH)

Selective panels of antibodies are used for cell surface markers to evaluate and diagnose PNH based on the clinical diagnosis and other primary lab findings.

Markers used for PNH Diagnosis:

FLAER (Alexa Fluor 488nm), CD59, CD11b, CD33, CD14, CD45 on patient and healthy control

Gating strategy: CD59 and FLAER expression on patient neutrophils, monocytes and RBCs (FSC/SSC, CD45/SSC, CD33/SSC)

5-6 ml of Sodium heparin anti-coagulated (Green top) peripheral blood samples of patient and a healthy control.
B5074 CD34+ cells Enumeration

Enumeration of CD34 positive cells in PBSCT and BMT samples

Gating Strategy: ISHAGE gating strategy – CD34 kit based enumeration

1mL of PBSC Samples / 5-6 ml of peripheral blood in sodium heparin anticoagulated (green top) tubes.


The following test is under validation and will be introduced soon in the laboratory

  • Dihydrorhodamine (DHR) flow cytometric test for Chronic Granulomatous disease (CGD)