Hello Kapish Gupta

You can use the Live/Dead viability assay kit from Invitrogen( Cat# L3224) with fluorescence microscopy which provides a two-color fluorescence cell viability assay with calcein-AM and ethidium homodimer-1. You can incubate the cells with 10uM calcein-AM and 10 uM ethidium homodimer-1 solution at 37 degree C for 40 minutes in the dark. Live cells are stained green and dead cells are stained red which can be imaged using fluorescence microscopy. Background signals are low because both dyes are non-fluorescent prior to interacting with cells. You can include areas in five to six different fields and then calculate percentage viability.

I hope this is helpful.

Best Wishes.

Hi, check out Promega, they have a new product line specifically designed to work in 3D cultures/spheroids. That might be close to your cells being encapsulated in matrigel.

Hello, you can try the AlamarBlue assay, but the CellTiter Glow ATP assay from Promega to me is best.

you could do a simple viability assay based on size based on the organoids you are growing. 50 um lower limit is a fairly standard size for these assays. Then count the organoids larger than your lower limit under a microscope or with an automated system.

https://www.oxford-optronix.com/resources/quantifying-organoid-size-and-counts

Hy Kapish Gupta

you can try with Nutral red uptake test (NRU) as they've done here Article An organoid culture assay (OCA) for determining the drug sen...

here is the protocol

https://cellculture2.altervista.org/neutral-red-dye-uptake-viability-test-nru/

good luck