I want to know if I can use AlamarBlue for determination metabolic activity of cells without dissolving alginate microspheres.
Hi Maxim,
Absolutely! You can do it directly without dissolving the capsules. We use routinely both CCK-8 (colorimetric) or Alamar Blue (fluorescence). This is a tentative protocol for you:
Between 1,000 and 25,000 cells per well are required to perform this assay (Alamar Blue is considerably more sensitive). Process:
o Measure 20 μl sedimented caps in small eppendorf. (It largely depends on the initial cell load in the pre-gel alginate solution. These calculations are for 5 million cells/mL).
o Carefully aspirate/remove supernatant.
o Add 500 μl of culture medium.
o Gently resuspend the capsules and pipette 100 μl / well into 96-well plate (x5).
o Add 10 μl of CCK-8/Alamar Blue reagent to each well.
o Incubate 1-4 h at 37ºC.
o Read
Hope this helps!
Edorta, thank you very much for your protocol. It's information is very helpful.
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