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Posted: Thu, 16 Jan 97 23:07:01 -0500
Date: Thu, 16 Jan 97 23:10:01 -0500
From: "Sam Witherspoon" <"witherspoon~s%Organization=Pharmacology%Telephone=(919)483-3078 FAX 483-3777"@a1.usa>
To: cyto-inbox
Subject: RE: Sorting insect cells and other fragile cells
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Hi Tony,
I sent the attached message to Geoff.
I think the best thing you can do is to increase the nozzle diameter if at
all possible. This will force a lower sheath pressure (and rate). You
won't get a reasonable break-off distance if you don't lower the sheath
pressure AND lower your drop drive frequency.
The draw-back is that it forces a lower drop rate. I have had good
luck with a 100u nozzle on a FACStar Plus. The drop drive produced @ 18
kHz.
Cheers,
Sam
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------- Forwarded message
Posted: Thu, 16 Jan 97 18:57:01 -0500
Date: Thu, 16 Jan 97 23:10:01 -0500
From: "Sam Witherspoon" <"witherspoon~s%Organization=Pharmacology%Telephone=(919)483-3078 FAX 483-3777"@a1.usa>
To: cyto-inbox
Subject: RE: Sorting insect cells?
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Hi Geoff,
I have sorted relatively fragile (and large) melanocytes from frog
skin. (:>).
I generally try to reduce the nozzle pressure and I think you will
accomplish this when you go up in nozzle size. On the FACStar Plus, with a
MACROSORT drop drive, we can get a "sortable" drop formation from a 70u
nozzle with a sheath pressure of 9 PSI, using a drive frequency of 29 to 30
KHz.
BTW, if these cell are infected, aren't they destined for death? Have
you considered single cell sorts to 96w plates? If you are concerned about
keeping cells (or contents) separate, this might be a solution.
Cheers,
Sam Witherspoon sw11527@glaxo.com
_________________________________________________________________
Flow Cytometry and Cell Sorting Resource
Dept. of Receptor Biochemistry
Glaxo Wellcome R&D.
RTP, NC 27709
Tel: 919-483-3078 FAX: 919-483-3777
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