Hi,
I am planning to detect viable Circulating Tumor Cell (CTC) by IF and downstream experiments. But how can I do IF without fixation and permeabilization since they'll harm the cells? Is it possible?
Thanks in advance.
If you have antibodies directed against surface makers you can do FACS analysis or magnetic bead extraction.
FACS analysis won't work as CTCs are vey low in number, if you can attach the cells to a surface lets say Poly-L-lysine you can to staining of the living cell without harming it, taking care and including blocking agents as BSA or comercial ones... I would say that you can fix the cell to perform IF without problem, otherwise it might be difficult to keep its integrity and you may loose the few cells that you have. What I know is colleagues doing CTC research they fix the cells with fixative and then spin the cells using a device called Cytospin, then they perform the IF in the slide. All the best.
Thank you Dr. John. I am using immunomagnetic beads for CTC isolation.
Thanks for your suggestion, Dr. Francisco. If I do cytospin, then don't I need to use fixative?
Dear Faysal,
We have a range of products that are designed for the analysis of CTC and rare cells in general. Using flow cytometry or cytospinning are only possible if the number of CTC is high. In general the number of CTC are somewhere between 0 - 5 per ml of blood which is too low to use cytometry or cytospining. To detect these low numbers you will some method to enrich the CTC. Our technologies are based on filtration followed by immuno fluorescence labeling and fluorescence imaging. The technology is designed to enumerate as well to isolate single CTC and is simple to use. Please take a look at our website www.vycap.com.
Best regards,
Arjan
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