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Trypan Blue Solution (0.4%)
Cell culture tested
Catalog Number LS 014-01
Storage Temperature 15~30°C
Although the aims of in vitro animal cell culture are various, cell counting is a common essential technique to every researcher. Especially, the viability of cells, i.e. the percentage of living cells among the total cells, is required for the next experimental steps. Trypan blue is a dye used in the living-cell counting. While living cells exclude this dye continuously and do not get stained, dead cells cannot exclude this dye and get stained blue. Trypan blue has high affinity for proteins in culture medium, thereby leaving background dark. This problem can be solved by centrifugation and resuspension of cells in protein free media or buffers.
LS 014-01 contains 0.4% (w/v) trypan blue in 0.81% (w/v) sodium chloride and 0.06% potassium phosphate (dibasic), and is sterilized by 0.2 mm filter. If necessary, dilute this solution to 0.2% concentration in PBS (LB 004 series), DPBS (LB 001 series), EBSS (LB 002 series), or HBSS (LB 003 series).
Trypan blue solution should be stored at 15~30°C. Deterioration of the liquid may be recognized by (1) precipitate or particulate matter throughout the solution, (2) cloudy appearance, (3) color change, and/or (4) pH change. Product label bears expiration date.
Biological Performance Characteristics
The biological performance characteristics of trypan blue are tested as the viability testing (living cell counting) in a medium containing 10% FBS using an appropriate cell line(s). (1) To 0.3 ml of balanced salt solution (PBS, DPBS, EBSS, or HBSS), add 0.2 ml of cell suspension and 0.5 ml of 0.4% trypan blue solution. (2) Mix thoroughly, allow it to stand for 5 min and observe/count the cells through a microscope. It should be noted that extended period may lead to absorption of dye by living cells.
For In Vitro Use Only