I am evaluating the viability and adhesion of cells on some different scaffolds (3d - printed, molded and electrospun). The lab making the electrospun scaffold for me adhered it to a large silicon wafer (too large for even a 24 well plate). In trying to plan out some initial experiments I was wondering if the Alamar Blue would work? or if my sample would be too dilute to differentiate cell activity from general background. For reference the scaffold in roughly .5mm in diameter and could probably only hold a few ul of cell suspension.
Dear Joshua
I have worked with polymeric scaffolds in different shapes (electrospun fiber, films, 3d-scaffolds) and alamarblue works perfectly to determine the metabolic activities of cells seeded on these scaffolds.
In my case, the samples were also about 5 mm diameter and I was placing a drop of 20 ul on the top with around 5000 cells. Once cells (HeLa) were attached (2 h), I added media to the well.
After 1 or 3 days of incubation, I changed the media and added media containing 10% of alamarblue. 4 hours of incubation were more than enough to get a good signal (in fluorescence). In any case, the incubation time can be increased up to 24 h to get more signal. The incubation time will depend on cell density and cell type.
Hope this helps.
Aitor
Aitor Larrañaga How much media were you adding to the well? My concern is that because of the size of the wafers the scaffolds are stuck to, I will have to place the wafers in a larger petri dish and use 2-3 mls of media to ensure the cells and scaffold are completely covered.
Hi Joshua, if you think that your sample might be too diluted you can increase the incubation time. I heard there is a new dye called PrestoBlue that is faster than the Alamar, but works on the same principle. It might be useful in your case.
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