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November 2003

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Subject:
Two photon
From:
Jason Goh <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Tue, 18 Nov 2003 10:25:29 +0000
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Hi all,

At the moment I'm designing a variant on a two-photon fluorescence microscope and as an optical engineer, have a question on a practical aspect of the operation.  

I'll be using fluorescein (FITC - excitation peak 494nm) as my fluorophore which would suggest a two-photon excitation wavelength of 988nm.  In the literature, I've not seen anyone use an excitation wavelength this high - mainly seems to be <800nm.  Is there any practical reason for this?  Is it due to decreased laser efficiency outside the optimum running wavelength (approx. 800nm).  I would have thought it was always best to operate at the excitation peak of the fluorophore to get more emission for the same amount of incident power...

Thanks in advance for your help,
Jason

----------------------------------------------------------
Dr Jason Goh
School of Electrical & Electronic Eng.
University of Nottingham
TEL:  +44 (0) 115 951 5556
FAX:  +44 (0) 115 951 5616

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