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Anyone heard of cells spontaneously fluorescing red??


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4 replies to this topic

#1 SuMi

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Posted 17 September 2010 - 07:14 AM

I had a very weird experience yesterday. I was running my Raji B cells on the flow cytometer and there was a positive red fluorescent peak in my negative sample. I went back to my flask of cells and ran an aliquot through the flow and they too were positive for red fluorescence. I looked at the cells under the microscope and some of the cells are glowing red. These cells weren't labelled with anything and so I have no idea as to how this spontaneous fluorescing happened. Anyone have any ideas? The only explanation I have is that it could be something in the media causing this but I have never seen it before and my cells were washed and resuspended in PBS before I ran them on the flow.

#2 laurequillo

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Posted 17 September 2010 - 07:25 AM

You could try DMEM without phenol red...
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#3 virusfan

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Posted 17 September 2010 - 07:49 AM

Could it be autofluorescence?

#4 SuMi

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Posted 23 September 2010 - 03:38 AM

Could it be autofluorescence?



I thought of that but dismissed it as cells would normally autofluoresce in the green channel and this was the red channel. There was no fluorescence in the green channel. I could try growing them without phenol red but I have been using these cells for two years and I've never had this problem before so I can't see how the medium could be the problem

#5 BNodes

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Posted 25 May 2011 - 10:28 AM

Cells autofluoresce in all channels. More importantly, I've noticed that when cells are stressed or when they apoptose the produce some fluorescent red chemical/protein. It should be relatively faint, and you should be able to separate them out with FSC and SSC gates.

-Bill


Could it be autofluorescence?



I thought of that but dismissed it as cells would normally autofluoresce in the green channel and this was the red channel. There was no fluorescence in the green channel. I could try growing them without phenol red but I have been using these cells for two years and I've never had this problem before so I can't see how the medium could be the problem






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