Nuclei abosolute counts

Hello flow people !

 

I would like to use a 'count' bead such as 'flow count' or similar to get an absolute count of Hoeschst stained breast cancer cell-line nuclei. 

 

If anyone is using count beads to get absolute counts by flow for nuclei preparations, it would be great to know what works well for you.  

 

If anyone sells count beads that could work, I would like to hear about it also.

 

I think I will need a count bead that can be excited by the violet laser to produce a signal around 450nm. This is because I may need to set a detection threshold using the hoechst signal (with a 450/50 emission filter and excitation off a violet laser) to see the nuclei if I cannot get a clear scatter signal to set a threshold on. 

 

I will use a CyAn ADP for this and I think one can only trigger on a single parameter here. Is this correct?

 

Any comments or advice or suggestions very much appreciated. 

 

Thanks so much for all the helpful responses that come out of this list, 

 

Best, 

Joanna 

 

 

 

Flow Cytometry Facility

AI 0129 	    EPFL

SV-SG		   Lausanne

Bâtiment AI	    Switzerland

 

+41 21 69 39 547