Re:Set up for Hoechst cell cycle analysis

From: Arnold Pizzey (a.pizzey@ucl.ac.uk)
Date: Tue Jun 13 2000 - 07:32:02 EST


At 10:46 12/06/00 +0000, you wrote:
>
>Dear Flow group,
>
>I am currently having problems setting up a FACS Vantage for Hoechst Cell
>cycle analysis.  It has an Innova 70 laser with UV maximum at 50mW.  I
>would like to know what beads or set up methods others have used to
>optimise the settings to get tight CV's.  I have tried using UV beads from
>Molecular Probes and then Calf thymus nuclei from the BD DNA QC particles
>box with 5 ug/ml Hoechst, however, I cannot get any decent looking cell
>cycle phases - I can only presume there are better methods than this
>attempt.  If anyone has any suggestions, it would be much appreciated.
>
>Regards
>Natasha.
>
>Natasha Webb
>
>Assistant Flow Cytometry Operator
>Room 5027
>Clinical Research Building
>Hammersmith Hospital
>London
>
>Phone:  0181 383 8330
>
>
>
>
Hello Natasha,

Try fixed peripheral blood lymphocytes;
Fix with 70% methanol @-20'C for one hour.
Aspirate off the methanol with a vacuum line.
stain.
These should give a nice tight G0 peak.

Alternatively, give me a call on the number below

best regards


Arnold


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	Arnold Richard Pizzey
	Department of Haematology
	Royal Free and University College London Medical School
	98 Chenies Mews
	London WC1E 6HX
	U.K

	voice:	+44 020-7679-6234
	Fax:	+44 020-7679-6222
	email:	a.pizzey@ucl.ac.uk
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