Do I need to convert 3D tumorsphere into single cell suspension before the alamarBlue treatment in assay? Or else can I use the intact tumorsphere directly for the alamarBlue treatment? Are there any related published protocols?
Dear Janith,
You can follow this recent paper. It may help you. PFA
Article Optimized alamarBlue assay protocol for drug dose-response d...
Dear Janith,
Maybe it would be a good idea to compare results of a viability assay with intact vs. dissociated spheres to be sure that you accurate results. The main issue with spheroids is the diffusion of compounds into the core – be it test compounds or reagents to test the viability – and the question is: do the compounds really reach the centre or are you only measuring the effect on the outer layer? Or in this case: Does the increased incubation time allow the alamarBlue to reach the core or is it only converted by the outer and maybe a few more deeper layers?
You could also try the following protocol, which uses Calcein AM and includes the dissociation of the spheroids, and compare the two methods:
https://www.promocell.com/product/3d-cell-culture-viability-assay-kit/
Hey Janith,
Why are you focusing on AlamarBlue as a viability assay? Are you open to using others? Promega has many products dedicated to 3D cell cultures. If I were you, I would have a look there and see if anything seems good. They even have samples for some of their products so you could test some for free too :
https://ch.promega.com/products/cell-health-assays/3d-cell-culture/
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