two bacteria populations observed when there should be just one - how to get rid of cell doublets? (Jul/07/2008 )

Helo!

We measure the amount of bacteria subpopulations. Sometimes we receive two spots in the dot blot diagram resulting that there are wrongly detected two subpopulations.

After some research through the internet, I found something about cell doublets.

Has anybody observed this strange results too?

How can I prevent this already in the sample preparation?

Thanks a lot!

Filtering the cells just prior to the flow cytometry analysis helps quite a bit. Also, use the smallest size filter you can.

I think a 40 micron filter is fairly standard, but depending on the size of the bacteria your are investigating you may want to find much smaller filters.

Also, check to see if the ions in your buffer improve or impair cell-cell adhesion.

Are you using EDTA in your FACS buffer?

May be you can filter'em?

Check these links if you find some tips on bacteria flow cytometry.

http://search.vadlo.com/b/q?sn=158621799&a...+flow&rel=0

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Hey, thanks a lot for your tips!
But honestly, I don´t know why filter them should prevent cell adhesions?

Hey, also thank you a lot for your answer!
Yea...EDTA is in the washing-buffer after hybridisation. But EDTA is a chelator...in what way it should influence cell adhesions?