Re: BD LSR optics

From: Geoffrey Osborne (geoff.osborne@anu.edu.au)
Date: Thu Sep 19 2002 - 18:44:46 EST


Dear Jim,
	We did quite a bit of work on this a while ago and came up with a solution
for 5 colour as per
http://jcsmr.anu.edu.au/facslab/facs/fitc-pe-tc-petx-apc.html
which is pretty easy to set up routinely. Note that the pin hole for the
filter block on P5 has to be inverted. If you want to go the whole hog and
do the APC-Cy7 (which we've used in this configuration
http://jcsmr.anu.edu.au/facslab/facs/fitc-pe-tx-apccy7.html
) in conjuction with the other 5 colours you mentioned then it's a whole
rethink on what you're doing. I couldn't figure out an easy way to do this
so we stuck with one fluorochrome coming off the UV line
http://jcsmr.anu.edu.au/facslab/LSRexample.html
which while not brilliant, with the right Ab/fl combination is usable as a
6 colour combination.
So there's my 2 cents worth. We had a the custom filters we needed made by
Chroma, who we really helpful, especially with the unusual AOI of some of
the filters.
Hope this helps
Geoff
---------------

At 10:27 AM 9/19/02 -0500, you wrote:
>
>To LSR users:
>
>Due to the cost of upgrade of the LSR to Digital electronics  and my need to
>use the system in 6 color mode ( FITC, PE, PE-TR, PE-Cy5, APC, APC-Cy7)  I
>need to go back to my original ideas of changing the filter setup of the
>instrument.
>I have made some progress about 6 months ago in doing this and was quite a
>task finding filters that would work at the odd angles the LSR is setup to
>use.
>If you have done this on your instrument and can do the above colors all at
>the same time please let me know.  No sense re-inventing the wheel if
>someone is able to share what they have learned.  I will be more than happy
>to share when I get all this working.
>
>Jim Houston
>
Geoffrey Osborne

Specialist, Flow Cytometry,
John Curtin School of Medical Research,
The Australian National University,
Canberra, 0200, ACT. AUSTRALIA
email: geoff.osborne@anu.edu.au
http://jcsmr.anu.edu.au/facslab/facshome.html

(61 2) 6125 3694.



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