RE: Cell Culture after DNA Ploidy Staining

From: Bob Ashcroft (cytomat@netcore.com.au)
Date: Thu Jul 17 1997 - 08:05:35 EST


Dear Leonie,

Sorry to hear of your problems, but there is a modestly priced solution, at 
least relative to getting a US$45,000++ laser to add to your sorter, along 
with the other costly detection and electronic items needed.

The choices depend on whether you wish to sort on the ploidy differences.

Hydroethidine is uptaken(??) by live cells and will bind DNA, though I 
don't know how good the histograms are. It excites at 488, and emits in the 
red, like ethidium.

Partec makes a PA, Ploidy Analyser, good for Ho33342 and DAPI, which 
definitely does the ploidy detection that you are looking for. It uses a Hg 
arc lamp. Their more advanced machines feature a 1,000 cells/s mechanical 
sorter, like the Calibur's 400/s sorter, but I don't know if that can be 
fitted to the PA.

Perhaps you should enquire of Dr Matthias Steinberg at info@partec.de. He 
is an experienced and helpful flow person.

Hope this helps; mail me for more; Bob

-----Original Message-----
From:	Leonie Gaudry [SMTP:l.gaudry@unsw.edu.au]
Sent:	Wednesday, July 16, 1997 10:48 AM
To:	Cytometry Mailing List
Subject:	Cell Culture after DNA Ploidy Staining


Hello,
I would like to ask if anyone has a method for estimating DNA Ploidy using
488nm or 633nm Laser so that the cells are alive and sortable before
culturing.If you use Hoechst and UV the above can be done,I am told.My
problem is no UV capabilities hence I am looking for some other way.
Hope someone out there can help me.
Thank you,Leonie



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