inter-beam compensation?

Joseph Webster J.Webster at centenary.usyd.edu.AU
Tue Nov 25 17:47:13 EST 1997


At 04:51 PM 23/11/97 -0500, f preffer wrote:
> Why is it necessary to have compensation [utilizing a FACScalibur] between
>FL4 and other fluor channels with 2 spatially distinct excitation sources?
>thanks
>frederic preffer

You are very fortunate that you CAN compensate between lasers!

You don't give details of your observations, but here's a guess - some
fluorochromes are excited by both 488 and 600+ lasers.

This effect is very obvious on dual laser sorters using a dye laser as
the second beam.
This *theoretically* should not be a problem in the FACScalibur, where
the second beam is longer wavelength (635) and low intensity like a few
milliwatts, but....

In our case, PerCP and the various tandem dyes are excited by the dye
laser and show up as false positives in either the APC or Texas Red
channel.

My question arising from this is:

Can someone tell me how to modify a FACStar Plus to compensate between
lasers?
We have the "7th active detector" modification, but can't find a usable
combination of 5 fluorochromes with 488 and dye laser!

	Thanks All, Joseph.

--
Joseph Webster
Flow Cytometry Facility
Centenary Institute



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