Flow Cytometry Group:
As a continuation of this thread by Gayle and Ray, a number of years
ago we ran into the problem of 3 or more colors overlapping. We worked with
our long-time collaborator Mark Corio to develop a high-speed three-color
compensation module for our sorter. Mark now sells a nice commercial version
of this system, which should work as a simple add-on (no problems about
warranty invalidations, etc)for just about any commercial or homebuilt flow
cytometer in the world, through his company. You would simply take the
outputs from the PMT preamps, pass them through this 3-color compensation
system (a box of software-controllable electronics), and plug the analog 0
to 1 volt outputs right back into the next step of your flow cytometer's
electronics. So other than inputting the proper compensation values,
everything runs exactly the same on your system as it always has. The system
has a nice software interface and runs off a PC (which is completely free to
do other things once it has "talked" to the hardware and downloaded the
color compensation values). In principle, two of these modules could be used
to do a 4- or 5-color compensation. We are now getting into situations where
this will likely be needed for our science.
I can vouch for the fine quality of Mark's work. He and I have
collaborated on many projects over the past ten years - and are still
collaborating on several projects. He is, in my opinion, one of the best
electronics designers in the field of flow cytometry. More detailed
information can be obtained at his website at http://www.rochestermicro.com.
I'm sure Mark would be happy to update you on his latest products. And if
what you want doesn't yet exist (my usual problem!), Mark can custom-build
it for you!
-- Best regards,
Jim Leary
-----Original Message-----
From: Ray Hicks [mailto:rh208@cus.cam.ac.uk]
Sent: Friday, November 26, 1999 4:06 AM
To: cyto-inbox
Subject: Re: Compensation for Green contaminating RED channel
Hi Gayle,
What kind of machine do you have? If it's a B-D one, there probably isn't
a direct FL3-%FL1 compensation circuit. You'll need to compensate through
the FL2-%FL1 and FL3-%FL2, ie compensate so that the signal is removed from
FL2 first, then compensate FL3 against FL2 until it disappears from FL3 as
well. If you're not using an FL2 reagent you might be able to skip the
first step.
Ray
At 10:41 am -0800 24/11/99, Gayle Hatleberg wrote:
>I am relatively new to the Flow field and I have been working with a
>Fluorescein like dye that seems to be contaminating our Red signal. The
>histogram of FL3 630/32 is very broad starting at 10^o and ending at
>10^3? Does anyone have any ides how to compensate out the signal. I
>have tried and the signal is not removed when compensating. Is there a
>good way to remove green signal that is contaminating the red channel?
>Thanks for your help
>Gayle Hatleberg
>Molecular Probes.Inc.
>Gayle@probes.com
Ray Hicks
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