I'd like to reply to Ray's e-mail with a quick note on automation.
Now we have been using the Cytek 96 well plate autosampler (called AMS)
for a few months with excellent results. The instrument is reliable, fast
and no steep learning curve or training is necessary to operate it. (The
AMS is running plates right now so I can write this message instead of
hunching over the Calibur.) The wash cycle between samples can be reduced
to a few seconds because the short sample line does not retain large
volume of cells. As a matter of fact a few seconds acquisition delay
prevents contamination better than long wash periods. Practically you can
go from the end of one sample to the start of the next one in 10-12
seconds with no contamination. The hardware is almost maintenance free and
you don't need to be present or supervise the AMS.
I only can hope that one day manufacturers start cooperating with each
other to the benefit of their customers and saving large development
expenses for themselves. Great hardware design combined with software
integration would be a winner. I am appealing to all cytometer
manufacturers here. Take a look at the AMS.
Akos (Biogen)
"Ray Lannigan" <lannigan@tritechinc.com>
11/07/2001 09:00 AM
Please respond to "Ray Lannigan"
To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
cc:
Subject: Re: FACS Calibur
Hi Carol,
There is an alternative to the 96 well plate autosampler that BD
offers.
It is called the Automatic Microsampler system and is made by Cytek
Development. If you would like more information please contact me.
As far as sorting on the Facscaliber, recovery of and viability of
sorted cells can be an issue. The inherent problem with the mechanical
sorting technique used in the FacsCaliber, is fluid constantly flows to
your
sort vessels, even when it is not trying to sort a cell in the sort gate.
After two hours of sorting, your 1000 - 2000 targeted cells could be in
approx. 300mL of fluid. The sorting mechanism is a catcher tube that goes
into the core stream of cells and attempts to catch the cell of interest.
When a cell is moving at a velocity of 6 meters/sec forcing a small tube
into its path, then forcing it to go through that tube can put relatively
extreme shear forces upon it. Stream-in-air with the electric charge
sorting
is the way to go.
Ray
-----Original Message-----
From: Carol Mazurek <mazurek@mpi.com>
To: cyto-inbox
Date: Tuesday, November 06, 2001 6:44 PM
Subject: FACS Calibur
>
>I realize that the FACS Calibur multiwell autosampler is fairly new, but
>is anyone actively using one? Could you please share your successes and
>failures with it? Does it do what you need? What would you change
>about it? Would you recommend it to others?
>
>Is there anyone with experience at sorting cells using the FACS
>Calibur? I realize it has limitations, and I'm not expecting FACS
>Vantage-quality sorting. I have a cell line that is expressing a
>recombinant protein and I want to sort the highest protein expressors
>(e.g., top 1% or 10%) in that population. What is the likelihood that I
>could get 1000 - 2000 targeted cells at >98% purity after sorting for an
>hour or two?
>
>
>Carol Mazurek
>Millennium Pharmaceuticals, Inc.
>mazurek@mpi.com
>
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