Description

 

Flow cytometry is a very fast and powerful method used to identify cells and classify them. In this method, the cells in the suspension are passed through a laminar flow from the flow cell and in front of the laser light, and in terms of physical characteristics such as size, granularity, shape and complexity of the nucleus, and also in terms of indicators on the surface of the membrane, the cytoplasm and nucleus by monoclonal antibodies that are labeled with fluorescence materials are investigated. In the flow cytometry section of The Payvand Laboratory, using four-color and six parametric flow cytometers, in addition to counting, determining the volume and granularity of cells, it can be used to study surface markers (CD markers) at the surface, cytoplasm or cell nucleus, and to use the results in order to diagnose, categorize, prognose and control the treatment of benign and malignant diseases, immunodeficiencies, and Paroxysmal Nocturnal Hemoglobinuria (PNH).

Important indicators of Flow Cytometry department are:

  • Establishing a quality management system in the framework of national and ISO standards (ISO 9001 + 9001)
  • Use four-color flow cytometry to increase sensitivity and detection characteristics
  • Use of International Standard Guidelines for Immunophenotyping for Acute and Chronic Leukemia
  • The first center in Iran to assess the minimum remaining disease (MRD) in patients under treatment for acute and chronic cancers of the blood.
  • Applying the standard high sensitivity method for diagnosis of anemia due to Paroxysmal Nocturnal Hemoglobinuria (PNH)
  • Evaluation and counting of lymphocytic subgroups in order to identify and categorize immunodeficiency diseases, infertility and ...
  • Evaluation of the involvement of body fluids including cerebrospinal fluid, pleura and ascites with malignant blood and non-blood cells, in accordance with international standards.
  • Preparing results in less than 24 hours and in emergency situations between 2-4 hours
  • Having the necessary experience and knowledge and getting scientific advice from university professors in time of need

Diagnostic Panels

 

1- Detection and classification of acute leukemia (Acute leukemia Flow cytometry panel)

In this panel, we detect acute types of leukemia including ALL (acute myeloid leukemia) and possible subgroups, by using monoclonal antibodies from the most reputable companies

The presence of the molecular and the FISH department facilitate the diagnosis of these cases according to the latest WHO classification. In addition, the report for each patient include the interpretive-diagnostic flow cytometry report, along with a cytomorphological report.

The following images selected from the results of the tests performed in the flow cytometry department of Payvand clinical and specialty laboratory:



 

2- Diagnosis and classification of chronic lymphoid malignancies (Lymphoid Neoplasm)

The detection of chronic B-T, and NK-like lymphoid malignancies including lymphocytic leukemia (CLL), hairy cell leukemia (Hairy Cell Leukemia) ), Mantel Cell Lymphoma, Spinal Lymphoma with Villous Lymphocyte, Follicular Lymphoma, Sezary Syndrome, Adult T-Cell Leukemia / Lymphoma, and many Other items are facilitated by using  cytomorphology, a complete panel of monoclonal antibodies (CD markers), molecular testing and FISH technique.

 

 

3- Determine the minimum residual disease in acute and chronic leukemia (Minimal Residual Disease (MRD) Acute & Chronic Leukemia)

Patients with acute and chronic leukemia have at least 13-12 malignant cells at the start of the diagnosis, followed by primary induction chemotherapy (Remission Induction), reduce to 10 or less, and the number of visible cells is reduced to a degree that morphology, would not be the valid method to be consider for evaluation , therefore, the use of higher sensitivity methods, including flow cytometry and molecular, would be considered.

For the first time in Iran, from 2012-2013, at Payvand clinical and specialty laboratory the MRD evaluation were made possible by use of flow cytometry, four colors and In addition, from 2018 onward the Payvand clinical and specialty laboratory equipped with BD Facslyrics next generation 10 color as well.

By Using the MRD results, high-risk patients can be distinguished from moderate or low risk patients, which are very effective in choosing treatment protocols. It is also possible to detect resistant cases and relapse sooner than the usual microscopic technique.

Determination of MRD in ALL children has been discussed since the 1990s and there are significant improvements in this area every day. In recent years, the MRD evaluation of the results in ALL adults has also been considered and recommended.

The MRD evaluation can also be used, For AML patients.

The minimum remaining disease in chronic leukemia like CLL also determines in recent years, and be considered in clinical trials, and it is be predicted to be added to therapeutic protocols for these patients, soon.

4- Use of flow cytometry to diagnose and monitor the treatment of patients with plasma malignancies, especially multiple myeloma

This test is common in many advanced centers in the world because by this method the detection of malignant cellular plasma become easier due to the changes in markers. After the initiation of treatment with this method, the MRD can be evaluate and control.

5- Flow cytometry of body fluids

The diagnosis of body fluid such as cerebrospinal fluid (CSF), pleural fluid, and ascitic fluid with malignant hematopoietic cells and non-tissue-derived tumors is one of the medical challenges.

CSF (neoplastic neuronitis) and other body fluids in lymphoma and acute leukemia’s occur despite preventive therapies, the conventional cell counting and microscopic evaluation methods do not have sufficient sensitivity to detect these cells, with 20-60% false negative usually reported in these methods. Therefore, by using a flow cytometry method, the diagnostic sensitivity can be increase, the type of cells and the malignancy origin can be determine, as well as increasing the value of diagnose.

 


6- Flow cytometry to evaluate the types of lymphocytes and count them in patients with innate and acquired immunodeficiency

Evaluation of innate and acquired immunodeficiency diseases, the assessment of the number and types of B, T, and NK-Cell lymphocytes, which, depending on the disease, which have been affected by one or more of these cells is one of the most commonly, asked tests.

For example, in HIV-infected patients, the reduction of the number of T-helper lymphocytes (Cd4 T helper) is one of the most prominent side effects of the disease that should be controlled.

 In patients with a variety of LAD, (Leukocyte Adhesion Deficiency), as well as the evaluation of CD16 and CD11a, 11b, 11c indexes on the neutrophils of the patient, makes diagnosis possible.

7- Evaluation of therapeutic antibodies with flow cytometry

flow cytometry is the method to identify surface antigens of cells that can be targeted for drug antibodies, After the treatment measure the responses to these medications, which are in fact monoclonal antibodies. Among the monoclonal antibodies that have a therapeutic aspect in hematologic and non-hematological diseases, in particular, autoimmune diseases (rheumatoid arthritis, multiple sclerosis, etc.), are the following:

-Anti - CD20 (Rituxan): B-cell malignant Lymphomas & Multiple Myeloma

-Anti - CD52 (Camputh-1 H): B-Cell Lymphocytic Leukemia & T-Cell Disorders

Measuring the expression of markers before treatment and at specific times after treatment should be done in order to evaluate the effect of treatment on normal cells.

In the interpretation of tests, the summary of the antigen expression pattern on the malignant cells as well as the normal cells that may be used in these antibodies should be considered.

8- Accelerated CT test to detect nightly attack (PNH) hemoglobin.

The best method for diagnosis of PNH is flow cytometry, which is performed by analyzing the indicators on granulocytes, monocytes and red blood cells with high sensitivity and potentiates the ability of the detection of PNH clones in these patients. In addition, the ability to detect PNH clones in patients with aplastic anemia and myelodysplastic syndrome (MDS), which is usually exist in low numbers, is possible.

 

 

 

 

 

Test List

No

Analyte/Investigation

Specimen type

Container Type /Volume

Transport /maintenance

Turnaround Time (TAT)

Emergency (STAT) case

Performed day

1

Adult T cell Leukemia Lymphoma
(ATL/ATLL) panel at diagnosis
( 16-18 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

2

ALL(B-cell) Body fluid panel
( 3-5 CD markers)

Serous Body Fluid: Pleural & Ascitic

EDTA lavender Cap / 5-10mL

4-8 °c

2 Days

Daily

Daily

3

ALL(B-cell) CSF panel for leukemic Meningitis
( 3-5 CD markers)

CSF

Sterile Universal Container- 3-5 mL

24 °c or room temperature

2 Days

Daily

Daily

4

ALL(B-cell) panel at diagnosis for MRD candidates
(24-26 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

5

ALL(B-cell) panel at diagnosis
(20-22 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

6

ALL(B-cell) panel for MRD
(20-24 CD markers)

Bone Marrow Aspirate (BMA)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

7

ALL(T-cell) CSF panel for leukemic Meningitis
( 3-5 CD markers)

CSF

Sterile Universal Container- 3-5 mL

24 °c or room temperature

2 Days

Daily

Daily

8

ALL(T-cell) panel at diagnosis
(20-22 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

9

ALL(T-cell) panel for MRD
(14-18 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

10

AML Body fluid panel
(4-6 CD markers)

Serous Body Fluid: Pleural & Ascitic

EDTA lavender Cap / 5-10mL

4-8 °c

2 Days

Daily

Daily

11

AML CSF panel for leukemic Meningitis
( 4-6 CD markers)

CSF

Sterile Universal Container- 3-5 mL

24 °c or room temperature

2 Days

Daily

Daily

12

AML panel at diagnosis
(22-28 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

13

AML panel for MRD
(16-20 CD markers)

Bone Marrow Aspirate (BMA)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

14

B or T cell NHL Body fluid panel
( 5-7 CD markers)

Serous Body Fluid: Pleural & Ascitic

EDTA lavender Cap / 5-10mL

4-8 °c

2 Days

Daily

Daily

15

B or T cell NHL CSF panel
( 5-7 CD markers)

CSF

Sterile Universal Container- 3-5 mL

24 °c or room temperature

2 Days

Daily

Daily

16

B ProLymphocytic Leukemia
(B-PLL) panel at diagnosis
( 14-16 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

17

CD34 for Stem Cell Enumeration

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB) /
Cord Blood

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

18

CLL panel at diagnosis
(14-16 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

19

CLL panel for MRD
( 8-12 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

20

DNA Index at Diagnosis

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

21

Eosin-5′-Maleimide (EMA) flowcytometry test for Hereditary Elliptocytosis

Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

22

Eosin-5′-Maleimide (EMA) flowcytometry test for Hereditary PyroPoikilocytosis

Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

23

Eosin-5′-Maleimide (EMA) flowcytometry test for Hereditary Spherocytosis

Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

24

Follicular Lymphoma panel at diagnosis
( 12-15 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

25

Hairy cell Leukemia panel at diagnosis
( 15-17 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

26

Hairy cell Leukemia panel for MRD
( 8-12 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

27

Lymphocyte Subset Enumeration
B-cell, T-cell & NK cell
(8-12 CD markers)

Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

28

Mantle Cell Lymphoma panel at diagnosis
( 14-16 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

29

Mantle Cell Lymphoma panel for MRD
( 8-12 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

30

NK Large Granular Lymphocyte (LGL) Leukemia panel at diagnosis
( 16-18 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

31

Plasma Cell Leukemia panel at diagnosis
( 12-16 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

32

Plasma Cell Leukemia panel for MRD
( 8-12 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

33

Plasma Cell Myeloma panel at diagnosis
( 12-16 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

34

Plasma Cell Myeloma panel for MRD
( 8-12 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

35

PNH Panel
Granulocyte/Monocyte & Erytrocyte
(10-14 CD markers)

Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

36

Sezary Syndrome panel at diagnosis
( 16-18 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

37

Splenic lymphoma with villous lymphocytes (SLVL) panel at diagnosis
( 15-17 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

38

T ProLymphocytic Leukemia panel at diagnosis
( 16-18 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

39

T-ALL Body fluid panel
( 3-5 CD markers)

Serous Body Fluid: Pleural & Ascitic

EDTA lavender Cap / 5-10mL

4-8 °c

2 Days

Daily

Daily

40

T-cell LGL Leukemia panel at diagnosis
( 16-18 CD markers)

Bone Marrow Aspirate (BMA) / Peripheral Blood (PB)

EDTA lavender Cap / 3mL

4-8 °c

2 Days

Daily

Daily

 

                                                                                                                                                                                                                                                                                            

Images

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                

                                                                                                                                                                                                                                                                                       

 

Comments  

#1 صمدی 2017-07-17 07:14
با سلام و تشکر

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