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Hi Jeff,
> I believe Hiro meant "polarizer at emission side", not excitation side.
Yes. I meant emission side. I was thinking to put this after scanner
where excitation and emission share their path, so it will work as
emission side.
Hiro.
>
> Other possible solutions:
> 1) If the scanhead allows one to add a laser-line 'notch' filter on the
> emission side, but preferably physically connected to the matching dichroic,
> this should work.
> 2) New emission filters, if they are old; and/or doubling them.
>
> Cheers,
> Jeff
>
> > -----Original Message-----
> > From: Hiroyuki Hakozaki [mailto:[log in to unmask]]
> > Sent: Tuesday, August 16, 2005 4:18 PM
> > To: [log in to unmask]
> > Subject: Re: Reflected light
> >
> >
> > ---------------------- Information from the mail header
> > -----------------------
> > Sender: Confocal Microscopy List <[log in to unmask]>
> > Poster: Hiroyuki Hakozaki <[log in to unmask]>
> > Subject: Re: Reflected light
> > --------------------------------------------------------------
> > -----------------
> >
> > Search the CONFOCAL archive at
> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> > Hi Carol,
> > If you put polarizer and quoter wave plate in optical path, you might
> > be able to prevent reflected excitation light.
> >
> > Put polarizer at excitation side and polarizer axis to be same
> > direction with laser polarization if laser is polarized. Locate quoter
> > wave plate for laser wavelength after polarizer - sample side. Quoter
> > wave plate axis need to be 45 degree to polarizer axis to create
> > circular polarization. Reflected laser light has reverse
> > rotation. After
> > passing quoter wave plate, it will be polarized and it is
> > perpendicular
> > to original laser polarization and blocked by polarizer.
> >
> > Drawback of this method is that You will lose more than half of your
> > fluorescence signal because of polarizer.
> >
> > We have Radiance2000 from Bio-rad and it has quoter wave plate and
> > polarizer in the optical path as default. I'm not familiar
> > with Leica system.
> >
> > Hiro.
> >
> > > Search the CONFOCAL archive at
> > > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> > >
> > > I run a core facility and several of the users grow their cells on
> > > silicon chips, which are very reflective. If they try to
> > see red and
> > > green fluorescence in the cells with our Leica SP2 confocal system,
> > > the reflected light is so bright that the fluorescence is
> > > overwhelmed. We tried closing the pinhole to .65 airy disk and that
> > > helped, but the background is still high and signal is
> > lost. It also
> > > helped to switch to the triple dichroic, relative to the double.
> > > Interestingly, with our old Bio-Rad MRC 600 and our 1024, the
> > > reflected light is not a problem. However, the signal is
> > low and the
> > > noise is much higher on these systems.
> > > Is there anything I can do to reduce the reflected light on the
> > > Leica? Why the difference between the systems?
> > >
> > > Thanks,
> > > Carol
> > >
> > > --
> > > <><><><><><><><><><><><><><><><><><><><><><>
> > > Carol Bayles, Manager
> > > Microscopy, Imaging & Fluorimetry (MIF)
> > > Biotechnology Resource Center
> > > 160a Biotech Bldg
> > > 607-254-4860
> > > www.brc.cornell.edu
> > >
> > > Confocal and Multiphoton Microscopy
> > > Nanobiotechnology Center
> > > www.nbtc.cornell.edu
> > >
> > > Cornell University
> > > Ithaca NY 14853
> >
> > =======================================
> > Hiroyuki Hakozaki
> >
> > National Center for Microscopy & Imaging Research
> > University of California San Diego
> >
> > Address:
> > 9500 Gilman Drive
> > Basic Science Building #1000
> > La Jolla, CA 92093-0608, USA
> > Tel: 858 534-2583
> > Fax: 858 534-7497
> > mailto:[log in to unmask]
> > NCMIR web site: http://ncmir.ucsd.edu
> > =======================================
> >
=======================================
Hiroyuki Hakozaki
National Center for Microscopy & Imaging Research
University of California San Diego
Address:
9500 Gilman Drive
Basic Science Building #1000
La Jolla, CA 92093-0608, USA
Tel: 858 534-2583
Fax: 858 534-7497
mailto:[log in to unmask]
NCMIR web site: http://ncmir.ucsd.edu
=======================================
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