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Flow cytometry (FC) is a technique used to detect and measure the physical and chemical characteristics of a population of cells or particles. [1] [2] [3] [4]In this process, a sample containing cells or particles is suspended in a fluid and injected into the flow cytometer instrument.
The whole procedure can be performed on cells from the blood, bone marrow or spinal fluid in a matter of a few hours. [citation needed] Immunophenotyping is a very common flow cytometry test in which fluorophore-conjugated antibodies are used as probes for staining target cells with high avidity and affinity.
The CD4 + /CD8 + ratio in the peripheral blood of healthy adults and mice is about 2:1, and an altered ratio can indicate diseases relating to immunodeficiency or autoimmunity. [2] An inverted CD4 + /CD8 + ratio (namely, less than 1/1) indicates an impaired immune system .
Lymphocytosis is usually detected when a complete blood count is obtained. If not provided the lymphocyte count can be calculated by multiplying the total white blood cell (WBC) count by the percentage of lymphocytes found in the differential count. [13] The lymphocyte count can also be directly measured by flow cytometry. [citation needed]
PIG-A assays were developed for cells of peripheral blood, such as red blood cells (RBCs) and white blood cells (WBCs). Due to conservative nature of GPI biosynthesis in mammalian species, similar flow cytometry protocols were developed for mammalian species of toxicological interest, i.e., mice and rats. [2]
The combination of the microscopic examination of the peripheral blood and analysis of the lymphocytes by flow cytometry to confirm clonality and marker molecule expression is needed to establish the diagnosis of CLL. Both are easily accomplished on a small amount of blood.