Fura2 imaging with Olympus TIRF system

[Andreas Bruckbauer]

Not a confocal topic but maybe someone knows:
I want to image fura2 with 340/380 nm excitation using our Olympus TIRF
system (cellR system, IX81) in widefield mode. The problem is that using the
Hg/Xe burner in the MT20 lamp, i have 20 times more power at 380 than 340
out of the objective, but a more reasonable 3 times out of the fibre. I have
a UApo/340 objective on loan, which should be suitable for 340 nm. The local
Olympus representative mentioned that the mirror in the L-shaped illuminator
can cut off some of the UV light. My question: has anyone experienced such a
problem and is it possible to change the mirror in the illuminator to a UV
reflecting one? The lens in the TIRF part of the illuminator is suitable for
405 nm, what is the transmission at 340 nm? Any suggestions welcome.

[Cameron, Lisa]

If you are going to use an optical fibre, you should also check what wavelengths
can be passed by the fibre. I know that 405 nanometers is on the edge of what my
fibre optic can pass.
Cheers,
Lisa


---------------------------------------
Lisa Cameron, Ph.D.
Director of Confocal and Light Microscopy
Dana Farber Cancer Institute
44 Binney St.; JF 215
Boston, MA 02115
Office phone: [phone]
Fax: [phone]
Office location: Jimmy Fund Bldg. 220B

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Andreas Bruckbauer
Sent: Friday, June 19, 2009 6:54 AM
To: [hidden email]
Subject: [CONFOCALMICROSCOPY] Fura2 imaging with Olympus TIRF system

Not a confocal topic but maybe someone knows:
I want to image fura2 with 340/380 nm excitation using our Olympus TIRF
system (cellR system, IX81) in widefield mode. The problem is that using the
Hg/Xe burner in the MT20 lamp, i have 20 times more power at 380 than 340
out of the objective, but a more reasonable 3 times out of the fibre. I have
a UApo/340 objective on loan, which should be suitable for 340 nm. The local
Olympus representative mentioned that the mirror in the L-shaped illuminator
can cut off some of the UV light. My question: has anyone experienced such a
problem and is it possible to change the mirror in the illuminator to a UV
reflecting one? The lens in the TIRF part of the illuminator is suitable for
405 nm, what is the transmission at 340 nm? Any suggestions welcome.


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