Hello again, Does anyone have experience analysing cells disaggregated from synovial tissue? I have a colleague who has been trying to do surface phenotyping of cells in synovial tissue without much success. A cell suspension was obtained using collagenase and DNase and the cells were stained with FITC conjugated CD3 and CD14. The results vary with different tissue samples but the gist of it is that the discrimination between negatives and positives is very poor with very dim FITC fluorescence. The same antibodies and staining protocol produce beautiful bright fluorescence with PBL's. Also, the collagenase has no effect on fluorescence intensity of CD3 or CD14 of the PBL's. Any suggestions that may help with this problem would be gratefully received. Cheers, Laura Dash.
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