http://confocal-microscopy-list.275.s1.nabble.com/Perkin-Elmer-OPAL-7-kit-experiences-tp7588991p7588998.html
quenching between the dyes. That said, we routinely do 6-8plex with
the imaging to a great degree. The bigger struggle is getting somewhat
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>
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>
> oh, wow, now we have a vendor that is using some value in the
> neighborhood of the actual peak of the *emission* to name dyes, just to
> add to the general confusion.
>
> Is there any reason to prefer the Opal fluors over those of vendors that
> freely publish the spectra of their dyes? Cost, maybe? From the maxima
> of the emission/excitation (not easily found at
>
>
http://www.perkinelmer.de/lab-products-and-services/application-support-knowledgebase/opal/opal-multiplex-ihc-assay.html),
>
> it does not seem to be something that would contain properties not
> available elsewhere.
>
> For those of you who have seen the spectra, do they look comparable to
> other dyes, concerning e.g. spectral width, or is there something that
> the vendor wants to hide?
>
> Steffen
>
>
> Am 12.12.2018 um 18:01 schrieb
[hidden email]:
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> >
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> > *****
> >
> > Hi Dave,
> >
> > one of our confocal users here has been tinkering around with Perkin
> Elmer's OPAL kits, testing various multicolor labeling schemes. At our
> request, Perkin Elmer sent us spectral data on the OPAL 7 dyes, albeit only
> as a png image file. For our purposes this was good enough, although proper
> data would have been a lot more useful. I'll send you the image of the
> spectra, if that would be of use to you (just shoot me an email).
> >
> > In general, you will need to do quite some spectral unmixing to get
> reasonable data when using some of the 'spectrally close' dyes. Any
> pre-separation of the dyes with excitation and detection multiplexing (such
> as was suggested with the 488nm & 514nm excitation for the 520 & 540 dyes,
> respectively) will go a long way towards acquiring useful data without
> incurring horrendous spectral uncertainty. But if you don't need all 7
> channels, best avoid the close ones entirely, if you have that option.
> >
> > Hope it helps!
> > Nicolai Urban
> > -----------------------------------------------------------------
> > Max Planck Florida Institute for Neuroscience
> > 1 Max Planck Way
> > Jupiter, FL 33458
> > -----------------------------------------------------------------
> >
> > -----Original Message-----
> > From: Confocal Microscopy List <
[hidden email]> On
> Behalf Of Dave Johnston
> > Sent: Mittwoch, 12. Dezember 2018 02:56
> > To:
[hidden email]
> > Subject: Perkin Elmer OPAL 7 kit experiences
> >
> > *****
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> >
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> > *****
> >
> > Hi All, I was wondering if anyone has had experience of using the Perkin
> Elmer & probe OPAL kit with confocal, especially with Leica spectral
> detection systems. With a fair amount of workup we have users using the
> OPAL 4 kit reliably but the OPAL 7 seems to provide significant extra
> challenges, particularly the very close overlap of the first 2 probes
> ("520" and "540" (actually 525 and 536 peaks) and the fact that no spectra
> seem to be available - the PE imaging systems would appear to rely on
> spectral unmixing using propriatory data. We have generated partial emision
> spectra for the 4 colour kit by lambda scanning but those likely wouldn't
> be good enough for unmixing purposes, which we would rather not have to
> rely on, and don't have a means of generating excitation spectra. Our best
> suggestion to date is to use the 4 colour kit on 2 consecutive serial
> sections. Any other suggestions and thoughts gratefully received.
> > Dave Johnston, Biomedical Imaging Unit, Southampton.
>
> --
> ------------------------------------------------------------
> Steffen Dietzel, PD Dr. rer. nat
> Ludwig-Maximilians-Universität München
> Biomedical Center (BMC)
> Head of the Core Facility Bioimaging
>
> Großhaderner Straße 9
> D-82152 Planegg-Martinsried
> Germany
>
>
http://www.bioimaging.bmc.med.uni-muenchen.de>