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We repeated all staining combinations with non-treated cells as controls.
These staining combinations have all been used previously for feline research (19).
The investigator can choose the reagent combinations which give answers to various specific questions: e.g. antisera to light chains establish the monoclonality of lymphomas, whilst staining combinations for human T and Ia-like antigens are particularly useful in various immunoregulatory disorders.
We used the separately treated cells for different staining combinations.
More complex staining combinations using various immune cell markers, as well as their functional markers, are being developed in our laboratory.
In addition, different staining combinations confirmed that CD163pos DDC expressed CD16, CD206 and CD209, but neither CD207 nor CadM1 (data not shown).
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Following two washes with supplemented RPMI, splenocytes were counted and aliquots of 1×106 cells were made for each staining combination as well as for flow cytometry compensation controls.
Thus, comparison of enteric neuronal structures between supposed normal and megacolonic samples is best done by opposing results of studies based on the same staining combination as we used here.
'Fluorescence minus one (FMO)' controls based on the staining combination used for the experimental sample but in which one antibody was left out and replaced with its isotype control were also used to set sort gates correctly.
Numbers in the cross tables indicate the count of staining intensity combinations from two methods.
Pericytes, endothelial cells, and apoptotic cells were identified by staining with combinations of 2 or 3 antibodies.
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