Has anyone used trypan blue to look at cell viability in fibroblasts (HFF cells)? I find all cells seem to take up the dye making it difficult to decipher between live/dead cells.
Hi Collin,
We always used trypan in our viability studies ((http://www.nature.com/cddis/journal/v7/n3/full/cddis2015409a.html OR https://www.researchgate.net/publication/296698867_Dengue-induced_autophagy_virus_replication_and_protection_from_cell_death_require_ER_stress_PERK_pathway_activation) AND
https://www.researchgate.net/publication/260112146_MTORp70S6K_signaling_distinguishes_routine_maintenance-level_autophagy_from_autophagic_cell_death_during_influenza_A_infection
OR http://www.sciencedirect.com/science/article/pii/S0042682214000117). Are you planning on using trypan to check viability? If you are using trypsin maybe they are very sensitive, try shortening the digest time. Also, you can try other viability assays like MTT or alamar blue. Hope these help!
-ED
Article Dengue-induced autophagy, virus replication and protection f...
Article MTOR/p70S6K signaling distinguishes routine, maintenance-lev...
Hi Colin
Yes, you can use trypan blue to access fibroblast viability. But you should associate this method with another one, such as MTT or Live/Dead. I don´t know how long you allowed the dye incorporate in the cells. I think 3-5 minutes of exposition is sufficient to observe the cells.
Best regards
Ana Celeste
Thanks for your answers,
I am trying to lyse and homogenise the cells and want a quick measure of how many cells were still alive. I have since used trypan to view cells after trypsinisation and they stain well, however I think the act of scraping the cells for lysis is enough to cause a lot of cell death.
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