Cell screening technology is to help isolate and identify certain cells with specific characteristics. This technique is increasingly used in the life sciences and medical research. There are many cell screening methods available, which can be chosen according to the cell type. The process of using flow cytometry for sorting target cells has distinct advantages among the many methods. It provides excellent speed, depth, and scale for cell analysis.
In flow cytometry, the cells are dispersed in a fluid, which is made to flow so that each cell is analyzed optically. A mixture of cells is sorted into different cell populations according to the optical characteristics of each cell. There are many parameters that can be used to screen cell populations by this technique. Thus, the method allows for the classification and analysis of a large number of cell lines.
Here, we describe cell screening methods using flow cytometry and common protocols. You can optimize the details according to your experimental needs to make it an effective method for your cell screening.
|Solutions and Reagents
|Phosphate buffer (PBS), dilution buffer, staining buffer
|Fluorescent dye-coupled antibody, antibody dilution buffer, staining buffer, washing buffer
Choose the appropriate cell sample preparation protocol based on your starting sample. The final sample is a homogeneous single cell suspension. It should contain no clumps or dead cell debris and have a density of 106-107 cells per ml.
Perform dye labeling on prepared cell suspensions. Select antibodies against specific cell surface molecules are used. Then incubate the cells with fluorescent dye-coupled antibodies. Common staining protocols include intracellular antigen staining, surface antigen staining, etc. Specific protocols can be developed based on experimental needs. After incubation, wash the cells with buffer.
Resuspend the washed cells in the buffer. The labeled cells are then passed through a flow cytometer. The flow cytometer uses a laser to excite the fluorescent dye and a detector to measure the fluorescence intensity of the dye. Based on the fluorescence intensity, the flow cytometer classifies the cells into different groups.
After screening, target cell populations are collected in test tubes or plates. Next you can check cell purity or other cell analysis using microscopy or flow cytometry.
Collected cells need to be kept alive
Are you interested in flow cytometry screening cell protocols? Please contact us for all key points of the relevant protocols, including experimental design, cell sample preparation, antibody selection, and data analysis.
For research use only. Not intended for any clinical use.