Hi everybody, Here's maybe a tricky one: Suppose you got highly autofluorescent cells and the isotype control is in the 2nd-3rd decade. The same isotype control in low autofluorescent cells is under the 1st decade. Now suppose the signal for marker X on these high autofluorescent cells is also in 2nd-3rd decade, and exactly overlaps the isotype control. What's the correct interpretation of these results? 1) there's no difference between isotype control and marker X, so the cells are negative for marker X or 2) marker X is in the 2nd-3rd decade, so it's definitely positive regardless of isotype control background Another way to state the question: are fluorochrome fluorescence (be it Ag-specific or aspecific binding) and autofluorescence additive signals for the same cell? TX a lot for any clue! Karim Pulmonary Immunobiology Ghent University Hosp.
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