I am using Tyramide Signal Amplification (TSA) staining on tumor cells (human derived xenograft in mice) to stain tumors for a panel of six markers. I am interested in using this same assay to stain the cell lines *before* injecting into mice. I understand that with the multiple microwave steps of the protocol, cancer cell lines grown on a glass slide will most likely fall off. I was considering embedding the cell lines in matrigel (50%+) as an analogue to tissue. Any suggestions? Has anyone managed to do this?
I had decent experience sticking cells to slides for IHC style assays (including TSA) using a cytospin protocol.
I don't think staining your cells prior to injection will be of much use since most staining procedures are going to kill the cells (eg permeabilize the membrane...)
Matthew Sobo, sorry I guess that wasn't clear. I wont be injecting these stained cells into mice. Rather I want to look at how the staining changes in the cell lines before versus the tumors (which are generated from injecting non-stained cells). Sorry for the confusion. What does the cytospin protocol entail?
Meredith Brown It is essentially a 'slow' centrifuge (a cytocentrifuge) that has a specially designed insert systems for pushing cells in solution (what depends on the requirements) onto a (typically + charged) glass slide. Filter paper absorbs the liquid during the spin as the cells are 'mounted' on the slide.
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