Hi everyone! does anybody have a clue if a parasite stained with CFSE dye dies inside macrophages, will CFSE staining be still detected? or only live parasites fluoresce? How do you minimize high problems of auto fluorescence from macrophages, while it's worst when infected with CFSE stained parasites?. Compensation is too hard.
Thank you for your valuable suggestions
To your first question, yes, the CFSE fluorescence will still bet detected if a parasite dies inside a macrophage. However if the parasite is taken up in acidic vacuoles, then the pH may inhibit the fluorescence of the CFSE. To your second question, there has been reports of using Trypan Blue on highly autofluorescent cells to reduce this background. You can try to search for keywords Trypan Blue and Autofluorescence to find some references.
Thank you very mych Ryn Dggn. I will try to find out for your suggested keywords.
Can you try switching to a CSFE alternative? Since macrophages autoflouresce green, might help to distinguish by using a different emission/excitation like violet or far red. Would be dependent on your instrument capability though.
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