RE: sorting marine microorganisms

We have experimented with using high salinity (~ 2 M) sheath to examine
Halo in our inFlux.  It works quite well, but we do have to do a long
cleaning and rinsing to remove all salt deposits.  Salt does
occasionally build up on the nozzle tip and disturb our stream during
the experiment.  On the inFlux it is easy to pause and remove it before
returning to the experiment.  I do not have experience with the Aria.
As an aside, the deflection on the charge plates does need to be turned
way down, and the drop position is quite different, so calibrating the
first time took some time.

 

Sincerely,

Gigi

Proteomics and Flow Cytometry Facilities

Institute for Systems Biology

1441 N. 34th St.

Seattle, WA 98103

206.732.1464 (ph)

206.732.1299 (fax)

  _____  

From: thomas.delohery@verizon.net [mailto:thomas.delohery@verizon.net] 
Sent: Thursday, September 14, 2006 7:20 PM
To: cyto-inbox
Subject: Re: sorting marine microorganisms

 

We used the equivalent of 5X PBS as sheath fluid in a FACS IV to purify
blood cell subsets from tunicates - see reference below.  I had the same
concerns you mention when I was approached with the project and was
surprised how well the instrument behaved.  It has been eons but I do
seem to recall that I had to use higher sheath pressures than normal due
to the higher density.	I have no direct experience with the Aria but
can not imagine why "artificial seawater" would be a problem.  2.7% of
what?

Regards, TomD

Oltz EM; Pollack S; Delohery T; Smith MJ; Ojika M; Lee S; Kustin K;
Nakanishi K
Distribution of tunichrome and vanadium in sea squirt blood cells
sorted by flow cytometry.
Department of Chemistry, Columbia University, New York, New York 10027.
Experientia 1989 Feb 15;45(2):186-90



Morrow, Matthew wrote:



All,

 

I have a potential client interested in sorting marine microorganisms.
Does anyone have experience doing this with a FACSAria? Are there any
effects on the instrument using a high-salinity, "artificial" seawater
sheath fluid (2.7%)? I'm concerned about salt crystal deposits in
valves, the nozzle, and on the grounding plate. Any feedback is
appreciated.

 

Cheers,

Matt

 

Matthew Morrow, MS, MT (ASCP)

Director, Flow Cytometry Core Facility

Children's Research Institute

University of South Florida

St. Petersburg, FL

(727) 553-3515 (office)

(727) 553-1295 (fax)

mmorrow1@hsc.usf.edu