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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.
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]
A blood smear is made by placing a drop of blood on one end of a slide, and using a spreader slide to disperse the blood over the slide's length. The aim is to get a region, called a monolayer, where the cells are spaced far enough apart to be counted and differentiated.
Cytometers are the instruments which count the blood cells in the common blood test.. Cytometry is the measurement of number and characteristics of cells.Variables that can be measured by cytometric methods include cell size, cell count, cell morphology (shape and structure), cell cycle phase, DNA content, and the existence or absence of specific proteins on the cell surface or in the ...
The entire procedure, once preparation is complete, typically takes 10–15 minutes. If several samples are taken, the needle is removed between the samples to avoid blood coagulation. After the procedure is complete, the patient is typically asked to lie flat for 5–10 minutes to provide pressure over the procedure site.
Previously, this procedure involved preparing a peripheral blood smear and manually counting each type of cell under a microscope, a process that typically required a half-hour. A Coulter counter played an important role in the development of the first cell sorter, and was involved in the early development of flow cytometry. Some flow ...
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.