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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.
Cell cycle analysis by DNA content measurement is a method that most frequently employs flow cytometry to distinguish cells in different phases of the cell cycle.Before analysis, the cells are usually permeabilised and treated with a fluorescent dye that stains DNA quantitatively, such as propidium iodide (PI) or 4,6-diamidino-2-phenylindole (DAPI).
Cells are immunostained in solution using methods similar to those used for immunofluorescence, and then analysed by flow cytometry. [citation needed] Flow cytometry has several advantages over IHC including: the ability to define distinct cell populations by their size and granularity; the capacity to gate out dead cells; improved sensitivity ...
The tetramers are labeled with a fluorophore, allowing tetramer-bound T-cells to be analyzed with flow cytometry. [4] Quantification and sorting of T-cells by flow cytometry enables researchers to investigate immune response to viral infection and vaccine administration as well as functionality of antigen-specific T-cells. [5]
MHC pentamers have been used in the detection of antigen-specific CD8+ T cells in flow cytometry, [12] and are cited in over 750 peer reviewed publications , including several in the journals Nature [22] and Science. [23] [24] MHC pentamers can also be used in tissue staining, [25] and in magnetic isolation of antigen-specific T cells. [26]
The fluorochrome-based TUNEL assay applicable for flow cytometry, combining the detection of DNA strand breaks with respect to the cell cycle-phase position, was originally developed by Gorczyca et al. [4] Concurrently, the avidin-peroxidase labeling assay applicable for light absorption microscope was described by Gavrieli et al. [5] Since 1992 the TUNEL has become one of the main methods for ...
If the cells do not have high intracellular potassium and if intracellular sodium is low, then (1) the cell membrane may not be intact, and/or (2) the sodium-potassium pump may not be operating well. [6] [7] Flow cytometry using 7-Aminoactinomycin D (7-AAD), wherein a lower signal indicates viable cells. Therefore, this case shows good ...
Flow cytometers can be used to collect multiparameter cytometry data, but cannot be used to separate or purify cells. Fluorescence-activated cell sorting (FACS) is a technique for sorting out the cells based on the differences that can be detected by light scatter (e.g. cell size) or fluorescence emission (by penetrated DNA, RNA, proteins or ...
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