[Cytometry] Scanning optical filters
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Tue Apr 16 13:10:35 EDT 2013
Seeing Ed's article cited reminded me how invaluable it was to scan every piece of optic glass we could slip out of our instruments at Rockefeller.
We used a Biochrom scanning spectrophotometer and I made a little MSAccess databases that had all the filter info plus graphs of both transmission and OD. For the most part they were consistent, but a few were stinkers.
As Howard Shapiro has noted several times, the OD is the real measurement and you need a spec that will measure 6 logs. Transmission % is not as telling.
On Apr 15, 2013, at 4:56 PM, Kelly Lundsten <klundsten at biolegend.com> wrote:
> Hi Suzanne!
> How are you? You have a 550/40 or so right now for Krome Orange in that position? The BV510 will still be really strong with that filter but you are right, the peak is at 520-ish to be ideal. Since you don't have a trigon on the Gallios, it's just a position for BV421 and BV510 (right?), you can use a 520/40nm filter to get the most out of the BV510. We just released another 5 BV510 conjugates just today, so that list will grow fast and will also be conjugated to the harder to detect markers like CD197 and CD117, etc. If you were detecting the BV510 on a trigon/octagon so that the BV510 was wedged between BV421 and BV570, then you might not want a big 40nm bandpass since the BV570 spillover would be annoying.
> Our experience in testing the filters for all of the Brilliant Violets was an interesting one. Half that were sent were promiscuous to the laser light and didn't pass CST. In my experience, trial and error is the only way to sort out good filters from inadequate.
> -----Original Message-----
> From: cytometry-bounces at lists.purdue.edu [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Suzanne Mertens
> Sent: Monday, April 15, 2013 9:52 AM
> To: cytometry at lists.purdue.edu
> Subject: [Cytometry] Custom BV510 filter for Gallios
> I'm working to get a filter set optimized for Biolegends' BV510. Does anyone have any special experiences doing this? I've done this before on an LSRII for the various PE tandems, so no big deal to work with Chroma - I just wanted to hear if anyone else has done this for the Gallios/Navios.
> PS. This is a shout-out to Ed Podniesinski: Your ICCS newsletter contribution on optical filters is awesome!
> - suzanne
> suzanne.mertens at gmail.com
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