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What causes autofluorescence? - (Feb/24/2011 )

Hi.

Today I noticed that cells from a mutant strain that I am using are more autofluorescent than the wild type. At first sight I thought of this as a big problem, but now I was wondering what can be leading the mutant cells to be more autofluorescent.

Importantly, I am talking about living, non-fixed cells.

Any ideas?


Thanks in advance.

-cardosopedro-

Can someone help me with this? :unsure:

-cardosopedro-

Definitely not an expert on this, but do your mutants contain amino acids that are being read in your channel that the wild type does not?

-mouse murderer-

Thank you for the reply. What do you mean? Amino acids in the medium? No, I use the same culture medium for both strains.

-cardosopedro-

I guess you are not working on skin cells, like melanocytes or keratinocytes? Or any other pigmented cell or tissue? Melanin is a great source of autofluorescence!
Apart from that, any big, organized structure can cause it, like Ribosomes, Actin filaments... Maybe you can help us by saying which cells you are working on and in which channel you find the increase.

-Rsm-

Hi again and thanks.

I work with fungal cells and I find increased autofluorescence in all channels of the cytometer.

-cardosopedro-

I am not a fungus expert, but doesn't Chitin show fluorescence all over the spectrum?

-Rsm-

I don't know... :( Is it?

-cardosopedro-