Usefulness of 1040-1080 nm in 2P Excitation

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Mayandi Sivaguru Mayandi Sivaguru
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Usefulness of 1040-1080 nm in 2P Excitation


Hi all, I have seen articles using 1030 nm Ex for mCherry, but the Coherent site showing data that it is best excited with 1060 nm, how significant the range 1040-1080 nm apart from mCherry in your experience for any other probes. In my personal experience, attempts to tune the laser closer to low and high end of the tuning range (730-930nm, particularly trying to obtain an excitation profile), in a previous version of Chameleon often tend to send the laser out of mode lock and requires a 15-30 min PZT recovery.

Thanking you in advance for your insights in this regard
Shiv


Mayandi Sivaguru, PhD, PhD
Microscopy Facility Manager
8, Institute for Genomic Biology
University of Illinois at Urbana-Champaign
1206 West Gregory Dr.
Urbana, IL 61801 USA

Office: 217.333.1214
Fax: 217.244.2496
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Periasamy, Ammasi (ap3t) Periasamy, Ammasi (ap3t)
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Re: Usefulness of 1040-1080 nm in 2P Excitation

Dear Shiv

Is your Verdi is 10W? If you have 10W pump power you should be able to auto-tune it easily in the range of 720-980nm. Yes, it is true that it may be noisy about 20nm on both ends for 10W pumping. If it’s a manual tuning and if you are an expert, you have lot of options to make the laser stable and mode locked.

I think we discussed in the email server before, most of the red fluorophores works pretty good between 740-760nm. We are using 740 for mCherry, mOrange, mKate, mKO and tdTomato, provide very good signal with our Biorad Radiance2100 with Nikon 60xIR lens. We use only few microwatts average power at the specimen plane. Of course double the wavelength will be nice.

Hope this helps

ammasi

 

 

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Mayandi Sivaguru
Sent: Friday, October 17, 2008 3:53 PM
To: [hidden email]
Subject: Usefulness of 1040-1080 nm in 2P Excitation

 


Hi all, I have seen articles using 1030 nm Ex for mCherry, but the Coherent site showing data that it is best excited with 1060 nm, how significant the range 1040-1080 nm apart from mCherry in your experience for any other probes. In my personal experience, attempts to tune the laser closer to low and high end of the tuning range (730-930nm, particularly trying to obtain an excitation profile), in a previous version of Chameleon often tend to send the laser out of mode lock and requires a 15-30 min PZT recovery.

Thanking you in advance for your insights in this regard
Shiv

 

Ammasi Periasamy, Ph.D.

Director, Keck Center for Cellular Imaging (KCCI)

Professor of Biology and Biomedical Engineering

Biology, Gilmer Hall (064), McCormick Rd

University of Virginia

Charlottesville, VA 22904

Voice: 434-243-7602 (Office); 982-4869 (lab)

Fax:434-982-5210; Email:[hidden email]

http//:www.kcci.virginia.edu

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