Howdy all,
I have stained tissue (double staining, DAPI + 488 nm antibody), which I want to break down so I can run FACS on it.
Wondering if anybody has experience with a procedure of this kind, and could please advise me on methods for breaking down tissue into single cells.
Thanks!
Hi Tzach
Can you specify what tissue? what amount of tissue to be processed? This might affect the protocol you need.
Boaz
Tzach, in the case of unfixed tissue, Collagenase is kind of standard. I'd simply give it a try. In worst case, you have to repeat the staining.
We've used collagenase digestion on tissue (about a half hour 37 degree shaking) then gently pressed the tissue through a falcon filter with a spatula.
https://www.fishersci.com/shop/products/falcon-cell-strainers-4/p-48680
Thank you all for your answers.
The tissue I wish to break down is a small amount (much less then a gram) of fixed insect embryo.
Thanks.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology