Immunophenotyping aids in the diagnosis and classification of several diseases as well as certain blood cell malignancies, such as lymphoma and leukaemia.
Doctors utilise a lab test called immunophenotyping to recognise and categorise particular cells.
It entails applying antibodies to a sample of your blood cells. Proteins called antibodies bind to particular antigens, also known as markers, that are present on the surface of white blood cells. Physicians can use these markers to categorise the cells.
If your doctor suspects you have leukaemia or lymphoma, or if they have already diagnosed you with one of these malignancies, they may conduct an immunophenotyping test.
What is the purpose of immunophenotyping?
Immunophenotyping gives medical professionals information to help them identify the precise type of cancer and the most effective treatment for each subtype.
For instance, it is capable of differentiating between the lymphocytes, which are white blood cells produced by your bone marrow. B cells, which stay in your bone marrow, and T cells, which go to your thymus, a lymphoid gland in front of your heart, are examples of lymphocytes.
This data is crucial for the diagnosis of malignancies like:
- acute myeloid leukemia (AML)
- erythroleukemia, a rare AML subtype
- B-cell or T-cell non-Hodgkin’s lymphoma
- B-cell or T-cell chronic lymphocytic leukemia (CLL)
- B-cell acute lymphoblastic leukemia (ALL)
- multiple myeloma
Immunophenotyping is also used by medical professionals to identify more aberrant cells that can point to illness and assess how well cancer treatments are functioning.
What are the types of immunophenotyping tests?
The following are the main immunophenotyping test types that physicians use:
- flow cytometry
- immunocytochemistry
- immunohistochemistry
Flow cytometry
The most popular method of immunophenotyping is flow cytometry. By determining whether particular antibodies cling to certain molecules on the surface of your white blood cells, it recognises molecules known as cluster of differentiation (CD) antigens or markers.
Your bone marrow or blood sample is placed in the fluid by lab personnel, who then treat it with fluorescent antibodies to stain your blood cells.
Subsequently, the sample is injected into a flow cytometer, an apparatus capable of analysing 10,000 cells per minute.
Cells from your sample are aligned into a single-file line using the flow cytometer. After that, a laser beam is focused on each cell, causing the fluorescent dyes to release light at particular wavelengths.
The quantity of light released reveals details about the cell, including its size and particular markers. This data is transformed into an analysis-ready format for computers.
A collection of recognised CD markers is updated and maintained by Human Cell Differentiation Molecules (HCDM). Among these markers are a few of them:
- B cell markers: CD19, CD20, and CD38
- Myeloid cell markers: CD11b, CD15, and CD33
- T cell markers: CD2, CD4, and CD7
Immunohistochemistry
Flow cytometry and immunohistochemistry (IHC) both use samples of blood cells that have been stained with fluorescent antibodies.
IHC, however, does not levitate the cell sample. Cell smears or tissue sections encased in paraffin, a waxy material, are used to analyse the cells.
IHC looks for certain antigens on the cells using a fluorescent or light microscope rather than a flow cytometer.
Immunocytochemistry
Although the terms "immunocytochemistry (ICC)" and "immunohistochemistry (IHC)" are sometimes used synonymously, they are not the same.
A fluorescent microscope is used by both IHC and ICC to assist in the identification of cells; however, ICC examines antigens in individual cells as opposed to the entire tissue sample.
While a biopsy is necessary for IHC, clinicians can frequently obtain a sample for ICC by a less invasive technique such as a smear or swab.
What is the procedure for immunophenotyping?
An immunophenotypist will first take a blood sample from a vein in your arm, or a physician may aspirate or biopsy bone marrow to get a tissue sample for immunophenotyping.
Doctors can use a blood sample for flow cytometry if prior tests, including a complete blood count (CBC), reveal aberrant cells in your blood. Compared to a biopsy or aspiration, it is less intrusive.
To perform an immunohistochemistry test, a tissue sample is required.
After that, a lab receives the blood or tissue sample for immunophenotyping.
What does an irregular immunophenotype result mean?
When immunophenotyping yields an irregular result, it suggests that aberrant blood cells are present because the CD markers on the surface of white blood cells don't fit the typical patterns.
Your age may have an impact on the outcome. Up until the age of 18, B cell and T cell numbers are typically lower in younger individuals. As people age, so does the way these cells' subgroups are broken down.
To ascertain the cause of an unusual immunophenotyping result, a physician may prescribe additional diagnostic testing.
FAQs
Are flow cytometry and immunophenotyping the same thing?
The most popular application of flow cytometry is immunophenotyping, or the identification of markers on cells, especially those related to the immune system. The rise of multicolor flow cytometry has been driven by the aim to identify more immune system cell subsets.
What’s the difference between immunophenotyping and immunohistochemistry?
The process of examining and categorising blood cells using antibodies and a fluorescent dye is known as immunophenotyping. One form of immunophenotyping is immunohistochemistry.
What’s the difference between immunohistochemistry and immunocytochemistry?
Both immunocytochemistry (ICC) and immunohistochemistry (IHC) examine cells for particular markers. On the other hand, ICC examines samples of cultured cells, and IHC examines tissue samples.
Takeaway
An aid in the diagnosis and classification of certain malignancies is immunophenotyping. This laboratory test looks for the presence of particular antigens that indicate disease using fluorescently stained blood or tissue samples.
These samples are analysed by flow cytometry, an immunophenotyping technique, with a laser beam, whereas immunohistochemistry makes use of a fluorescent microscope.
The outcomes of immunophenotyping assist a physician in creating a successful treatment strategy and prognosis for that particular illness.
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