FCM
steven4564 authored about 2 years ago
Simple Introduction
The full name of FCM is Flow Cytometry. We can also call it FACS whose full name is Fluorescence activated Cell Sorting. It's a novel analytical technique and separation technique for rapid measurement of cellular or subcellular structures. By the flow cytometer, it will make the single cell or other small biological particles in rapid straight flow state, and get through the light beam one by one, thus getting multiparameter (quantity, size, nucleic acid content, cell activity, specific bacterial colony or species, etc.) quantitative analysis and detection for individual cells or particles. It has the outstanding advantages of fast, sensitive, accurate and convenient to operate.
It has the following characteristics：1.High speed for measure. 2. Multiparameter measurement is available. 3. It is a comprehensive high-tech method (FCM combines optics, electronics, fluid mechanics, cytochemistry, immunology, lasers, computers and so on). 4. It is not only a cell analysis technique, but also a precise sorting technique.
FCM is commonly used in the following aspects:
- (1)To measure the mutation coefficient of DNA in cells through this technology was the lowest, usually less than 2%.
- (2)It can perform DNA ploidy analysis accurately.
- (3)With fluorescent dyes, quantitative studies on proteins and nucleic acids in cells are carried out.
- (4)Rapid cell sorting and cell collection.
- (5)Medical applications: Studies on immune function and detection of various stem cells, the multi-drug resistance of cancer patients, cell function and metabolic dynamics research, analysis of platelets (cardiovascular disease), flow cytometry and molecular biology research.
- (6)Being applied to the measure of peripheral blood endothelial cells, regulatory T cells and other cutting-edge fields.
Flow cytometer
Flow cytometryis a new kind of high-tech instrument whose core technology is flow cytometry, integrating fluid science, optics, electronics, biology and immunology. Facs services can quickly measure, store, display in a series of important characteristic parameters in biophysics and biochemistry of those cells which are suspended in the liquid dispersion. In addition, it has the ability to select the specified subgroups of cells according to the preselected parameters.
#
Flow cytometry is mainly composed of fluid flow system, optical system, signal collection and conversion system, analytical system and cell sorting system.
Fluid Flow System
Flow cytometry is mainly composed of fluid flow system, optical system, signal collection and conversion system, analytical system and cell sorting system.
Fluid Flow System
Optical System
#
Signal Collection and Conversion System(Lin)
#
Signal Collection and Conversion System(Log)
Fluorescence Activated Cell Sorting
By fluorescence activated cell sorting, flow cytometry analysis can quickly get the biological characteristics and biochemical components of single cell and quantitatively determine these parameters.
Cell fluorescence signal mainly includes two parts:
- autofluorescence, is the fluorescence which isn't produced by fluorescent molecules inside cells after exposure to light.
- Characteristic fluorescence, that is, by the fluorescent dye attached to the cell by dyeing, the fluorescence which is produced after exposure to light
The fluorescence signal which FCM detected and analyzed is mainly refers to the fluorescence signal emitted through specific fluorescence staining. It can maintain the structure and function of cells and organelles or particles without being destroyed. With the help of a fluorescent probe, from the molecular level, flow cytometry will acquire a variety of signal on quantitative analysis of cell separation or purification.
The application of flow cytometry has been widely used, a word can be summed up as, all cells or particles which can be fluorescent molecular marked could be detected by flow cytometry instrument, especially for the extremely tiny particles, the advantages of tests, rapid, sensitive and multiparameter analysis are more obvious.
In addition to its high initial cost, one of the key problems in FCM is the selection of fluorescent dyes and treatment of samples, which challenges the development of fluorescent dyes. In a word, with the rapid development of science and technology, flow cytometry will have a wider application prospect.