How to prepare sample for FACS using Frozen Tissue?

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Sang Ho Lee

Hello Rashimi.

I am afraid that the frozen-thawed tissues would not work for FACS. Even if you have a small proportion of cells, they probably lost their antigens. Collect fresh liver, get single cell suspension for your FACS. Without any cryoprotectants and serum, even freezing and thawing of dispersed cells will rupture and destroy organelles. Tissues are even more damaged.

Regards.

Langyue Hu

Hi, Rashimi. Please read the article carefully, the authors have developed a protocol for flow cytometry using frozen tissues. I hope it will be useful.Article A rapid and optimization-free procedure allows the in vivo d...

Hamid Yousf Dar

Hi Rashmi

I am also afraid that frozen sample will work for facs because cells get ruptured as already stated by Mr Lee.

I think you should fo FACS analysis of fresh spleen sample. You have to do RBC lysis to it before staining. You can see current immunology protocols for better understanding.

Good luck.