Edit* I am looking for a way to quantify how many cancer cells are in an organ. I was hoping to isolate all cells from the organ and then do FACS or a cell staining assay. What really matters is getting close to 100% recovery of cells from a whole murine organ and a method to count total starting cells and total/subgroups of recovered cells.Any recommendations or references?
I am currently searching for methods of quantifying metastasis other than histology and imaging. These cells would have to be in a functional/viable format (for flow cytometry or immuno techniques. I am currently reviewing chimeric organ and cancer metastasis literature.
Cut up organ in 4 degree C M199 or PBS without Ca and Mg. Add cold trypsin with EDTA overnight in the refrigerator. Remove from refrigerator and pipette up and down gently until the tissue falls apart. Pass the chunkies and cells through a 20 um screen and collect in 50 mL conical. Centrifuge at 1000 xg for 10min at 10 degree C. Aspirate supernatent with a pipette and resuspend cells in medium of choice.
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