Trypan Blue, also known as Trypan Blue, Trypan Blue, and Triphenyl Blue, is a cell viability dye that is often used to detect the integrity of cell membranes and cell survival. Trypan blue staining is one of the most commonly used dead cell identification staining methods in tissue and cell culture. Cells that have lost their activity or have incomplete cell membranes can be stained blue with Trypan blue, while normal living cells have intact cell membrane structures and can exclude Trypan blue, so the cells will not be stained blue. It is generally believed that the loss of cell membrane integrity means that the cell is dead. Macrophages can engulf Trypan blue, so it can be used as a vital stain for macrophages. In addition, it should be noted that apoptotic bodies also refuse to be stained with Trypan blue.
After Trypan blue staining, the cell survival rate can be more accurately quantified by directly counting under a microscope or taking pictures under a microscope. Staining only takes 3-5 minutes and is simple to operate.
Instructions for use
1. Collect cells (adherent cells may need to be digested with trypsin and/or EDTA), centrifuge at 1000 r/min for 5 minutes, discard the supernatant, prepare a single cell suspension, and dilute appropriately.
2. Add 0.4% trypan blue solution to the cell suspension, the final concentration of trypan blue is 0.04%, and stain at room temperature for 3 minutes (the staining time can be slightly extended, but should not exceed 10 minutes).
3. Take a small amount of stained cells and count them with a hemocytometer. Under a microscope, it can be observed that dead cells are blue and swollen, and dull; living cells are not stained and maintain normal morphology and are shiny.
4. Calculate the viability of live cells: Cell viability (%) = Total number of live cells/(Total number of live cells + Total number of dead cells) × 100%
Precautions
1. The staining time should not be too long, otherwise the live cells will gradually accumulate dyes and become colored, causing deviations in the test results;
2. When counting cells, it is usually necessary to quantify them more accurately, and each cell sample should be counted for no less than 500 cells;
3. This product is only used for scientific research or further research, not for diagnosis and treatment.
