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Near Infrared Imaging

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Michal Jarnik

Near Infrared Imaging

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Hi Listers,

A user in my facility needs to image in near infrared (tissue sections)
- excitation of 750 nm, emission 775 nm. Does anyone have experience in
this sort of imaging? As for a light source, merkury arc lamp seems to
be out of question and even xenon does not emit there very strongly.
What are the other options? I would expect Nikon to provide a
corresponding filter set, but is xenon lamp strong enough? Or what else
one could use (short of retrofitting our confocal with a near IR laser,
which would probably be possible).

Any insight would be very much appreciated.

Thanks, Michal

--

Michal Jarnik, Ph.D.
Cell Imaging Facility
Electron Microscope Facility
Fox Chase Cancer Center
333 Cottman Ave.
Philadelphia, PA 19111

Craig Brideau

Re: Near Infrared Imaging

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You can get fairly powerful LEDs at that wavelength if you want to do
widefield imaging. Otherwise if you want confocal you will need an
appropriate laser. Here's a bunch of plug-and-play LEDs for microscopy:
http://www.thorlabs.com/NewGroupPage9.cfm?ObjectGroup_ID=2615
The one that emits at 735 should work for you; LEDs are broad emitters so it
will definitely overlap into the 750nm range.

(no commercial interest -just like playing with LEDs)

Craig

On Tue, Nov 16, 2010 at 11:49 AM, Michal Jarnik <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Listers,
>
> A user in my facility needs to image in near infrared (tissue sections) -
> excitation of 750 nm, emission 775 nm. Does anyone have experience in this
> sort of imaging? As for a light source, merkury arc lamp seems to be out of
> question and even xenon does not emit there very strongly. What are the
> other options? I would expect Nikon to provide a corresponding filter set,
> but is xenon lamp strong enough? Or what else one could use (short of
> retrofitting our confocal with a near IR laser, which would probably be
> possible).
>
> Any insight would be very much appreciated.
>
> Thanks, Michal
>
> --
>
> Michal Jarnik, Ph.D.
> Cell Imaging Facility
> Electron Microscope Facility
> Fox Chase Cancer Center
> 333 Cottman Ave.
> Philadelphia, PA 19111
>

Guy Cox-2

Re: Near Infrared Imaging

In reply to this post by Michal Jarnik

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

You could use a multiphoton microscope as single-photon. It would be
pretty tricky to get the signal back to the confocal detectors but it
sounds like you don't need confocal imaging, so you could use a
non-descanned detector with some custom filters (which you're going to
need however you do it). PMTs aren't too crash hot at that wavelength
but they do still work.

Guy

Optical Imaging Techniques in Cell Biology

by Guy Cox CRC Press / Taylor & Francis

http://www.guycox.com/optical.htm
<http://www.guycox.com/optical.htm>

______________________________________________

Associate Professor Guy Cox, MA, DPhil(Oxon)

Australian Centre for Microscopy & Microanalysis,

Madsen Building F09, University of Sydney, NSW 2006

Phone +61 2 9351 3176 Fax +61 2 9351 7682

Mobile 0413 281 861

______________________________________________

http://www.guycox.net <http://www.guycox.net>

From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Michal Jarnik
Sent: Wednesday, 17 November 2010 5:49 AM
To: [hidden email]
Subject: Near Infrared Imaging

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Listers,

A user in my facility needs to image in near infrared (tissue sections)
- excitation of 750 nm, emission 775 nm. Does anyone have experience in
this sort of imaging? As for a light source, merkury arc lamp seems to
be out of question and even xenon does not emit there very strongly.
What are the other options? I would expect Nikon to provide a
corresponding filter set, but is xenon lamp strong enough? Or what else
one could use (short of retrofitting our confocal with a near IR laser,
which would probably be possible).

Any insight would be very much appreciated.

Thanks, Michal

--

Michal Jarnik, Ph.D.
Cell Imaging Facility
Electron Microscope Facility
Fox Chase Cancer Center
333 Cottman Ave.
Philadelphia, PA 19111

________________________________

No virus found in this message.
Checked by AVG - www.avg.com
Version: 10.0.1153 / Virus Database: 424/3259 - Release Date: 11/15/10

Jim Beacher

Re: Near Infrared Imaging

In reply to this post by Michal Jarnik

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To join, leave or search the confocal microscopy listserv, go to:
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*****

*** Commercial Interest ***

Hi Michal/ List

CoolLED can offer LED illumination at 740nm. LEDs remain powerful into
the NIR and we believe that you will have sufficient intensity. We would
be pleased to lend you a unit to test and compare with other solutions.

Feel free to contact me directly at [hidden email]

JIM Beacher

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Listers,
>
> A user in my facility needs to image in near infrared (tissue sections)
> - excitation of 750 nm, emission 775 nm. Does anyone have experience in
> this sort of imaging? As for a light source, merkury arc lamp seems to
> be out of question and even xenon does not emit there very strongly.
> What are the other options? I would expect Nikon to provide a
> corresponding filter set, but is xenon lamp strong enough? Or what else
> one could use (short of retrofitting our confocal with a near IR laser,
> which would probably be possible).
>
> Any insight would be very much appreciated.
>
> Thanks, Michal
>
> --
>
> Michal Jarnik, Ph.D.
> Cell Imaging Facility
> Electron Microscope Facility
> Fox Chase Cancer Center
> 333 Cottman Ave.
> Philadelphia, PA 19111
>

David Burk

Re: Near Infrared Imaging

In reply to this post by Guy Cox-2

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Please forgive me if this is a stupid question. How can one use their
multiphoton as a single-photon? Is there a way to operate, say, a Mai
Tai at 760nm (or any tunable wavelength) in CW mode? If so, this would
be pretty useful.

Thanks,
David

David H. Burk, PhD.
Cell Biology and Bioimaging Core
Pennington Biomedical Research Center
6400 Perkins Rd.
Baton Rouge, LA 70808

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Guy Cox
Sent: Wednesday, November 17, 2010 12:17 AM
To: [hidden email]
Subject: Re: Near Infrared Imaging

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

You could use a multiphoton microscope as single-photon. It would be
pretty tricky to get the signal back to the confocal detectors but it
sounds like you don't need confocal imaging, so you could use a
non-descanned detector with some custom filters (which you're going to
need however you do it). PMTs aren't too crash hot at that wavelength
but they do still work.

Guy

Optical Imaging Techniques in Cell Biology

by Guy Cox CRC Press / Taylor & Francis

http://www.guycox.com/optical.htm
<http://www.guycox.com/optical.htm>

______________________________________________

Associate Professor Guy Cox, MA, DPhil(Oxon)

Australian Centre for Microscopy & Microanalysis,

Madsen Building F09, University of Sydney, NSW 2006

Phone +61 2 9351 3176 Fax +61 2 9351 7682

Mobile 0413 281 861

______________________________________________

http://www.guycox.net <http://www.guycox.net>

From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Michal Jarnik
Sent: Wednesday, 17 November 2010 5:49 AM
To: [hidden email]
Subject: Near Infrared Imaging

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Listers,

A user in my facility needs to image in near infrared (tissue sections)
- excitation of 750 nm, emission 775 nm. Does anyone have experience in
this sort of imaging? As for a light source, merkury arc lamp seems to
be out of question and even xenon does not emit there very strongly.
What are the other options? I would expect Nikon to provide a
corresponding filter set, but is xenon lamp strong enough? Or what else
one could use (short of retrofitting our confocal with a near IR laser,
which would probably be possible).

Any insight would be very much appreciated.

Thanks, Michal

--

Michal Jarnik, Ph.D.
Cell Imaging Facility
Electron Microscope Facility
Fox Chase Cancer Center
333 Cottman Ave.
Philadelphia, PA 19111

________________________________

No virus found in this message.
Checked by AVG - www.avg.com
Version: 10.0.1153 / Virus Database: 424/3259 - Release Date: 11/15/10

Julio Vazquez

Re: Near Infrared Imaging

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To join, leave or search the confocal microscopy listserv, go to:
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*****

Hi David,

Single photon versus multi-photon excitation is primarily a matter of
photon density (illumination intensity). Otherwise, I don't think
molecules care too much about whether your laser is CW or pulsed, and
in that respect, the so-called "multiphoton laser" is a bit of a
misnomer. Therefore, you can probably use your MaiTai (or other)
pulsed laser to achieve conventional (single photon) fluorescence
excitation.... you may just need to bring down the power
considerably. Alternatively, it is possible to achieve two-photon
excitation with CW lasers, although in this case you need quite high
powers. see for instance:

Gu, Tannous and Shepard. Optics Communications 117: 406-412 (1995)

Booth and Hell. Journal of Microscopy 190: 298-304 (1998)

--
Julio Vazquez
Fred Hutchinson Cancer Research Center
1100 Fairview Ave. N., mailstop DE-512
Seattle, WA 98109-1024

Tel: Office: 206-667-1215/ Lab: 206-667-4205
FAX: 206-667-6845

[hidden email]
http://www.fhcrc.org/science/shared_resources/imaging/

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On Nov 18, 2010, at 10:10 AM, David Burk wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Please forgive me if this is a stupid question. How can one use their
> multiphoton as a single-photon? Is there a way to operate, say, a Mai
> Tai at 760nm (or any tunable wavelength) in CW mode? If so, this
> would
> be pretty useful.
>
> Thanks,
> David
>
> David H. Burk, PhD.
> Cell Biology and Bioimaging Core
> Pennington Biomedical Research Center
> 6400 Perkins Rd.
> Baton Rouge, LA 70808
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]]
> On Behalf Of Guy Cox
> Sent: Wednesday, November 17, 2010 12:17 AM
> To: [hidden email]
> Subject: Re: Near Infrared Imaging
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> You could use a multiphoton microscope as single-photon. It would be
> pretty tricky to get the signal back to the confocal detectors but it
> sounds like you don't need confocal imaging, so you could use a
> non-descanned detector with some custom filters (which you're going to
> need however you do it). PMTs aren't too crash hot at that wavelength
> but they do still work.
>
>
>
>
> Guy
>
>
>
> Optical Imaging Techniques in Cell Biology
>
> by Guy Cox CRC Press / Taylor & Francis
>
> http://www.guycox.com/optical.htm
> <http://www.guycox.com/optical.htm>
>
> ______________________________________________
>
> Associate Professor Guy Cox, MA, DPhil(Oxon)
>
> Australian Centre for Microscopy & Microanalysis,
>
> Madsen Building F09, University of Sydney, NSW 2006
>
>
>
> Phone +61 2 9351 3176 Fax +61 2 9351 7682
>
> Mobile 0413 281 861
>
> ______________________________________________
>
> http://www.guycox.net <http://www.guycox.net>
>
>
>
>
>
> From: Confocal Microscopy List
> [mailto:[hidden email]]
> On Behalf Of Michal Jarnik
> Sent: Wednesday, 17 November 2010 5:49 AM
> To: [hidden email]
> Subject: Near Infrared Imaging
>
>
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Listers,
>
> A user in my facility needs to image in near infrared (tissue
> sections)
> - excitation of 750 nm, emission 775 nm. Does anyone have
> experience in
> this sort of imaging? As for a light source, merkury arc lamp seems to
> be out of question and even xenon does not emit there very strongly.
> What are the other options? I would expect Nikon to provide a
> corresponding filter set, but is xenon lamp strong enough? Or what
> else
> one could use (short of retrofitting our confocal with a near IR
> laser,
> which would probably be possible).
>
> Any insight would be very much appreciated.
>
> Thanks, Michal
>
> --
>
> Michal Jarnik, Ph.D.
> Cell Imaging Facility
> Electron Microscope Facility
> Fox Chase Cancer Center
> 333 Cottman Ave.
> Philadelphia, PA 19111
>
> ________________________________
>
> No virus found in this message.
> Checked by AVG - www.avg.com
> Version: 10.0.1153 / Virus Database: 424/3259 - Release Date: 11/15/10

Mark Cannell

Re: Near Infrared Imaging

In reply to this post by David Burk

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi David

You just need to prevent mode locking. Since the cavity does not start
in a mode locked condition, if you can disable the starter mechanism
the laser will happily run in CW. If you also, reduce intracavity GVD
compensation the laser will be even less likely to mode lock if
bumped. I suggest you contact the Manufacturer on this.

Hope this helps.

Mark

On 19/11/2010, at 7:10 AM, David Burk wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Please forgive me if this is a stupid question. How can one use their
> multiphoton as a single-photon? Is there a way to operate, say, a Mai
> Tai at 760nm (or any tunable wavelength) in CW mode? If so, this
> would
> be pretty useful.
>
> Thanks,
> David
>
> David H. Burk, PhD.
> Cell Biology and Bioimaging Core
> Pennington Biomedical Research Center
> 6400 Perkins Rd.
> Baton Rouge, LA 70808
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]
> ]
> On Behalf Of Guy Cox
> Sent: Wednesday, November 17, 2010 12:17 AM
> To: [hidden email]
> Subject: Re: Near Infrared Imaging
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> You could use a multiphoton microscope as single-photon. It would be
> pretty tricky to get the signal back to the confocal detectors but it
> sounds like you don't need confocal imaging, so you could use a
> non-descanned detector with some custom filters (which you're going to
> need however you do it). PMTs aren't too crash hot at that wavelength
> but they do still work.
>
>
>
>
> Guy
>
>
>
> Optical Imaging Techniques in Cell Biology
>
> by Guy Cox CRC Press / Taylor & Francis
>
> http://www.guycox.com/optical.htm
> <http://www.guycox.com/optical.htm>
>
> ______________________________________________
>
> Associate Professor Guy Cox, MA, DPhil(Oxon)
>
> Australian Centre for Microscopy & Microanalysis,
>
> Madsen Building F09, University of Sydney, NSW 2006
>
>
>
> Phone +61 2 9351 3176 Fax +61 2 9351 7682
>
> Mobile 0413 281 861
>
> ______________________________________________
>
> http://www.guycox.net <http://www.guycox.net>
>
>
>
>
>
> From: Confocal Microscopy List [mailto:[hidden email]
> ]
> On Behalf Of Michal Jarnik
> Sent: Wednesday, 17 November 2010 5:49 AM
> To: [hidden email]
> Subject: Near Infrared Imaging
>
>
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Listers,
>
> A user in my facility needs to image in near infrared (tissue
> sections)
> - excitation of 750 nm, emission 775 nm. Does anyone have experience
> in
> this sort of imaging? As for a light source, merkury arc lamp seems to
> be out of question and even xenon does not emit there very strongly.
> What are the other options? I would expect Nikon to provide a
> corresponding filter set, but is xenon lamp strong enough? Or what
> else
> one could use (short of retrofitting our confocal with a near IR
> laser,
> which would probably be possible).
>
> Any insight would be very much appreciated.
>
> Thanks, Michal
>
> --
>
> Michal Jarnik, Ph.D.
> Cell Imaging Facility
> Electron Microscope Facility
> Fox Chase Cancer Center
> 333 Cottman Ave.
> Philadelphia, PA 19111
>
> ________________________________
>
> No virus found in this message.
> Checked by AVG - www.avg.com
> Version: 10.0.1153 / Virus Database: 424/3259 - Release Date: 11/15/10

Guy Cox-2

Re: Near Infrared Imaging

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

You can still do single photon excitation with a mode-locked laser. But
if you want to prevent two-photon excitation (eg of another dye) you can
set it to cw mode. On my old Coherent Mira you just pressed the CW
button (which disabled the starter) and opened the slit (so that Kerr
lensing didn't do anything). I'd imagine a manual Spectra-Physics
would have similar controls - on the new computer-controlled ones I'd
imagine it's in the software somewhere.

Guy

Optical Imaging Techniques in Cell Biology

by Guy Cox CRC Press / Taylor & Francis

http://www.guycox.com/optical.htm
<http://www.guycox.com/optical.htm>

______________________________________________

Associate Professor Guy Cox, MA, DPhil(Oxon)

Australian Centre for Microscopy & Microanalysis,

Madsen Building F09, University of Sydney, NSW 2006

Phone +61 2 9351 3176 Fax +61 2 9351 7682

Mobile 0413 281 861

______________________________________________

http://www.guycox.net <http://www.guycox.net>

From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Mark Cannell
Sent: Friday, 19 November 2010 8:11 AM
To: [hidden email]
Subject: Re: Near Infrared Imaging

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi David

You just need to prevent mode locking. Since the cavity does not start
in a mode locked condition, if you can disable the starter mechanism
the laser will happily run in CW. If you also, reduce intracavity GVD
compensation the laser will be even less likely to mode lock if
bumped. I suggest you contact the Manufacturer on this.

Hope this helps.

Mark

On 19/11/2010, at 7:10 AM, David Burk wrote:

[mailto:[hidden email]

> ]
> On Behalf Of Guy Cox
> Sent: Wednesday, November 17, 2010 12:17 AM
> To: [hidden email]
> Subject: Re: Near Infrared Imaging
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> You could use a multiphoton microscope as single-photon. It would be
> pretty tricky to get the signal back to the confocal detectors but it
> sounds like you don't need confocal imaging, so you could use a
> non-descanned detector with some custom filters (which you're going to
> need however you do it). PMTs aren't too crash hot at that wavelength
> but they do still work.
>
>
>
>
> Guy
>
>
>
> Optical Imaging Techniques in Cell Biology
>
> by Guy Cox CRC Press / Taylor & Francis
>
> http://www.guycox.com/optical.htm
> <http://www.guycox.com/optical.htm>
>
> ______________________________________________
>
> Associate Professor Guy Cox, MA, DPhil(Oxon)
>
> Australian Centre for Microscopy & Microanalysis,
>
> Madsen Building F09, University of Sydney, NSW 2006
>
>
>
> Phone +61 2 9351 3176 Fax +61 2 9351 7682
>
> Mobile 0413 281 861
>
> ______________________________________________
>
> http://www.guycox.net <http://www.guycox.net>
>
>
>
>
>
> From: Confocal Microscopy List

[mailto:[hidden email]

> ]
> On Behalf Of Michal Jarnik
> Sent: Wednesday, 17 November 2010 5:49 AM
> To: [hidden email]
> Subject: Near Infrared Imaging
>
>
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Listers,
>
> A user in my facility needs to image in near infrared (tissue
> sections)
> - excitation of 750 nm, emission 775 nm. Does anyone have experience
> in
> this sort of imaging? As for a light source, merkury arc lamp seems to
> be out of question and even xenon does not emit there very strongly.
> What are the other options? I would expect Nikon to provide a
> corresponding filter set, but is xenon lamp strong enough? Or what
> else
> one could use (short of retrofitting our confocal with a near IR
> laser,
> which would probably be possible).
>
> Any insight would be very much appreciated.
>
> Thanks, Michal
>
> --
>
> Michal Jarnik, Ph.D.
> Cell Imaging Facility
> Electron Microscope Facility
> Fox Chase Cancer Center
> 333 Cottman Ave.
> Philadelphia, PA 19111
>
> ________________________________
>
> No virus found in this message.
> Checked by AVG - www.avg.com
> Version: 10.0.1153 / Virus Database: 424/3259 - Release Date: 11/15/10

________________________________

No virus found in this message.
Checked by AVG - www.avg.com
Version: 10.0.1153 / Virus Database: 424/3263 - Release Date: 11/17/10

David Burk

Re: Near Infrared Imaging

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Thanks to everyone who has responded. I appreciate it. My confocal
representative contacted me and we are checking with our laser rep on
how to switch to CW from mode-locked. It never occurred to me to A)
just try exciting a near IR fluorophore with a mode-locked laser or B)
simply asking the manufacturer how to switch to CW. For some reason I
did not think this was an option. I guess my reasoning, or lack
thereof, can best be explained by this semi-analogous question - "Should
I expect to find cup holders in an Aston Martin?"

David

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Guy Cox
Sent: Thursday, November 18, 2010 3:38 PM
To: [hidden email]
Subject: Re: Near Infrared Imaging

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

You can still do single photon excitation with a mode-locked laser. But
if you want to prevent two-photon excitation (eg of another dye) you can
set it to cw mode. On my old Coherent Mira you just pressed the CW
button (which disabled the starter) and opened the slit (so that Kerr
lensing didn't do anything). I'd imagine a manual Spectra-Physics
would have similar controls - on the new computer-controlled ones I'd
imagine it's in the software somewhere.

Guy

Optical Imaging Techniques in Cell Biology

by Guy Cox CRC Press / Taylor & Francis

http://www.guycox.com/optical.htm
<http://www.guycox.com/optical.htm>

______________________________________________

Associate Professor Guy Cox, MA, DPhil(Oxon)

Australian Centre for Microscopy & Microanalysis,

Madsen Building F09, University of Sydney, NSW 2006

Phone +61 2 9351 3176 Fax +61 2 9351 7682

Mobile 0413 281 861

______________________________________________

http://www.guycox.net <http://www.guycox.net>

From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Mark Cannell
Sent: Friday, 19 November 2010 8:11 AM
To: [hidden email]
Subject: Re: Near Infrared Imaging

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi David

You just need to prevent mode locking. Since the cavity does not start
in a mode locked condition, if you can disable the starter mechanism
the laser will happily run in CW. If you also, reduce intracavity GVD
compensation the laser will be even less likely to mode lock if
bumped. I suggest you contact the Manufacturer on this.

Hope this helps.

Mark

On 19/11/2010, at 7:10 AM, David Burk wrote:

[mailto:[hidden email]

________________________________

No virus found in this message.
Checked by AVG - www.avg.com
Version: 10.0.1153 / Virus Database: 424/3263 - Release Date: 11/17/10

Kate Luby-Phelps

Re: Near Infrared Imaging

In reply to this post by Michal Jarnik

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I'd be interested to know if you can really operate the multiphoton laser in
CW. I was advised by my manufacturer not to try.

As an alternative we have been imaging dyes in this EX/EM range (especially
IR Dye800) with a standard wide-field scope and a Cy7 filterset from Chroma.
We have tried both the halogen lamp (switched to the epi illumination
position) or the Xenon. Both work surprisingly well. You might want to use a
back-thinned CCD with extended red sensitivity. We have been using the
Hamamatsu Orca BT 1024.

This arrangement won't work for very weak signals but for most
immunofluorescence applications we have been successful.

Kate Luby-Phelps

Craig Brideau

Re: Near Infrared Imaging

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If you can't unmodelock your laser, one thing you can do is fire it through
a big block of glass to stretch the pulses out into the many picosecond
regime. This will help keep 2 photon effects from happening so you can
concentrate on 'CW' effects. It's not perfect, but it is really easy to
implement. Just put a glass block between the laser and your microscope, an
inch thick ought to do it.

Craig

On Fri, Nov 19, 2010 at 6:57 AM, Kate Luby-Phelps <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I'd be interested to know if you can really operate the multiphoton laser
> in
> CW. I was advised by my manufacturer not to try.
>
> As an alternative we have been imaging dyes in this EX/EM range (especially
> IR Dye800) with a standard wide-field scope and a Cy7 filterset from
> Chroma.
> We have tried both the halogen lamp (switched to the epi illumination
> position) or the Xenon. Both work surprisingly well. You might want to use
> a
> back-thinned CCD with extended red sensitivity. We have been using the
> Hamamatsu Orca BT 1024.
>
> This arrangement won't work for very weak signals but for most
> immunofluorescence applications we have been successful.
>
> Kate Luby-Phelps
>

Guy Cox-2

Re: Near Infrared Imaging

In reply to this post by Kate Luby-Phelps

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As I said, I don't know about these computer-controlled ones, but on my
manual Mira I was advised always to put it in CW mode if I had to do a
substantial alignment, and only try to get it mode-locking when the beam
was back and fully tweaked. This was just because there are fewer
variables in CW. This has now been replaced with a Mai-Tai DeepSee
(installed last week) but I haven't yet had a play with it.

Guy

Optical Imaging Techniques in Cell Biology

by Guy Cox CRC Press / Taylor & Francis

http://www.guycox.com/optical.htm
<http://www.guycox.com/optical.htm>

______________________________________________

Associate Professor Guy Cox, MA, DPhil(Oxon)

Australian Centre for Microscopy & Microanalysis,

Madsen Building F09, University of Sydney, NSW 2006

Phone +61 2 9351 3176 Fax +61 2 9351 7682

Mobile 0413 281 861

______________________________________________

http://www.guycox.net <http://www.guycox.net>

From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Kate Luby-Phelps
Sent: Saturday, 20 November 2010 12:57 AM
To: [hidden email]
Subject: Re: Near Infrared Imaging

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I'd be interested to know if you can really operate the multiphoton
laser in
CW. I was advised by my manufacturer not to try.

As an alternative we have been imaging dyes in this EX/EM range
(especially
IR Dye800) with a standard wide-field scope and a Cy7 filterset from
Chroma.
We have tried both the halogen lamp (switched to the epi illumination
position) or the Xenon. Both work surprisingly well. You might want to
use a
back-thinned CCD with extended red sensitivity. We have been using the
Hamamatsu Orca BT 1024.

This arrangement won't work for very weak signals but for most
immunofluorescence applications we have been successful.

Kate Luby-Phelps

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