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Hi David and Mark,

Good points. By specialized laser and optics, I meant a UV argon laser and UV quartz optics. I had such a beast once to image NADH, but multiphoton is much easier and more versatile.

Regarding multiphoton photodamage, David Piston has a paper showing that 2 photon imaging is more damaging than 1 photon imaging for cells in culture and that the multiphoton photodamage has a 3 photon power profile. Hence most folks will tell you to use conventional 1 photon imaging for cells in culture -- but you'll still need 2-photon microscopy to image things like NADH and NADPH unless, of course, you have a UV laser and UV optics.

John

--
John J. Lemasters, MD, PhD
Professor and GlaxoSmithKline Distinguished Endowed Chair
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Medical University of South Carolina
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of David Knecht charter
Sent: Saturday, April 23, 2011 6:03 PM
To: [log in to unmask]
Subject: Re: Live cell imaging with multiphoton confocal

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Is there any disadvantage to multiphoton for cultured cells (besides cost and complexity)? Cell viability?  Phototoxicity?  Dave

On Apr 23, 2011, at 11:21 AM, Kate Luby-Phelps wrote:

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> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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> 
> Multiphoton has no clear advantage for cells in culture. For cell 
> culture samples, we use two photon  only to excite DAPI or other UV 
> and near-UV fluors since we had to make a choice between the Ti:S and a 405 laser on our scope.
> 
> Kate Luby-Phelps

Dr. David Knecht    
Department of Molecular and Cell Biology Co-head Flow Cytometry and Confocal Microscopy Facility
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)