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Hi-
We'd be using one or the other but I can imagine a case where you'd want
to do one TIRF experiment followed (or preceeded by) a confocal
experiment on the same sample/ field of view. I've heard that
polarization issues may be important, but I'm not interested in any
anisotropy experiments, so I don't really understand why that would be
the case.
Sarah
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Holly Aaron
Sent: Tuesday, June 01, 2004 12:22 PM
To: [log in to unmask]
Subject: Re: Fiber optic splitter?
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Hi, Sarah -
I have not done this (yet), but at least in theory it is possible. It
probably depends on the power of your laser.
I *think* that TILL Photonics offers such a solution "off the shelf"
more or less, but don't know for sure. Check out:
http://www.till-photonics.com/products/accessories/tirf1.html
We have been talking to McBain Instruments
[http://www.mcbaininstruments.com/] about sharing lasers for spinning
disk confocal (more laser required than for laser scanning applications)
and TIRF. They are working out a beam-splitting option which would
direct percentage of total laser to one application or the other (90/10
to 10/90). Do you want to be able to do both simultaneously or would you
be using only one OR the other? Lastly, alignment may always be an
issue if switching applications.
Best of luck to you!
-Holly
__________________________
Holly L. Aaron
Imaging Specialist
Molecular Imaging Center
Cancer Research Laboratory
447 Life Sciences Addition #2751
University of California Berkeley
Berkeley, CA 94720-2751
510.642.2901
510.642.5741 fax
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http://imaging.berkeley.edu
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Michael Cammer
Sent: Tuesday, June 01, 2004 7:36 AM
To: [log in to unmask]
Subject: Re: Fiber optic splitter?
Search the CONFOCAL archive at
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A lot depends on the efficiency of the fluorescent probe and the S/N of
the detector. For instance, with TIRF 0.2 mW 488 nm at the back
aperture of objective is fine for our samples that can involve long (>
2s) exposure times but inadequate for dim signals. 2.5 mW may be fine
for small granules moving quickly in living cells (but exposure times of
approx. 100ms which is still too slow for some applications), but the
trade off is bleaching or killing the cells. -Michael C.
At 10:28 AM 5/28/2004 -0400, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Hi-
>We have a confocal that we are planning to add TIRF capabilities to.
>Would it be possible to share our Ar/Kr laser between the two inputs
>with a fiber-optic splitter(i.e. half light going to the scan head,
>half to the TIRF module)? I'm new to TIRF so I'm not sure what sort of
>laser power one needs. It seems like for confocal at least we have
>laser power to spare, so losing half of it wouldn't be a big deal. Any
>thoughts about this?
>Thanks-
>Sarah
>
>-----------------------------------------------------------------------
>-
>---------------
>Sarah Locknar, Ph.D.
>Director, Neuroscience COBRE Imaging / Physiology Core
>College of Medicine, University of Vermont
>E015 Given Building
>89 Beaumont Ave.
>Burlington, VT 05405
>802-656-0413
>802-656-8704 (fax)
>-----------------------------------------------------------------------
>-
>---------------
________________________________________________________________________
____
Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of
Med.
Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY
10461
(718) 430-2890 Fax: 430-8996 URL:
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