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Thank you for you very useful comment.
I will keep your experience with FAD into close consideration as I  
think it is relevant with us. I have a lot of experience with imaging  
yeast and there too leaving cells on plates too long will increase  
autofluorescence.
Thank you
Caterina


On 26-ott-07, at 17:07, Sandrine Pouvreau wrote:

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> Hi. I'm not sure wether what I will say is relevant in your case,  
> but I have had problem with autofluorescence in my cells while  
> willing to use the FITC channel. This fluorescence is colocalised  
> with mitochondria. I did a spectrum of it, and it matches with  
> emission spectrum of FAD. This autofluorescence is greater in cells  
> that have been in the dish for a while (probably due to oxidation,  
> FAD is fluorescent in the oxidised state).
> I'm not sure that this case apply to your HeLa cells (I never  
> worked with them), but I hope it will be of some help
> Best
> Sandrine Pouvreau
>
> Rush University
> Chicago, Il
>
>
>
>
> Caterina Strambio <[log in to unmask]>
> Sent by: Confocal Microscopy List <[log in to unmask]>
> 10/26/2007 08:09 AM
> Please respond to
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> cc
> Subject
> HeLa autofluorescence
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> Hi
> we want to use adherent cells (ideally HeLa cells) for live imaging  
> of viral
> particles traveling to the cell nucleus. We are having a lot of  
> problems
> with  autofluorescence in the Green/FITC channel. It is not simply  
> diffuse
> autofluorescence but it has a clear morphology: green dots all over  
> the
> cytoplasm.
>
> We are growing cells in phenol red free DMEM and it did not help  
> all that much.
>
> Any suggestions?
>
> Thank you very much for your help.
>
> Caterina Strambio De Castillia
>
> Istitute of Research in Biomedicine
> Bellinzona
> Switzerland
>