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P.S. UV-transmitting optics don't have to be quartz. Many lens used for UV illumination of Fura2 or Indo1 transmit UV but are still not chromatically corrected in the UV as is desirable if not necessary for point scanning confocal. 

--
John J. Lemasters, MD, PhD
Professor and GlaxoSmithKline Distinguished Endowed Chair
Director, Center for Cell Death, Injury and Regeneration
Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular Biology
Medical University of South Carolina
QF213 Quadrangle Building
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Charleston, SC 29425

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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Lemasters, John
Sent: Saturday, April 23, 2011 9:21 PM
To: [log in to unmask]
Subject: Re: Live cell imaging with multiphoton confocal

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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 Director, Center for Cell Death, Injury and Regeneration Departments of Pharmaceutical & Biomedical Sciences and Biochemistry & Molecular Biology Medical University of South Carolina
QF213 Quadrangle Building
280 Calhoun Street, MSC 140
Charleston, SC 29425

Office: 843-792-2153
Lab: 843-792-3530
Fax: 843-792-8436
Email: [log in to unmask]
http://academicdepartments.musc.edu/ccdir


-----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)