Changing system from NIR (two-photon) to UV
Locked
 
|
G. Esteban Fernandez |
Changing system from NIR (two-photon) to UV
|
Hello all,
On our Zeiss LSM 510 META two-photon system we have a Coherent Chameleon NIR laser that may need major repair. Coherent has been very good in providing remote technical assistance to us but we're at a point where a service visit is required and we're not under contract. Since we do more shallow imaging of UV dyes like DAPI on that system that two-photon imaging, we would replace the NIR laser with UV if it is more cost-effective than repairing the NIR. Zeiss says a switch from NIR to UV would require a major (and pricey) overhaul of the optics. I don't know what that entails but it is understandable given the huge difference in wavelength. However, I wonder how badly out of focus or aberrated the UV image would actually be with the optics that are already in place if we put a UV laser in there in place of the NIR; we do have a UV laser dichroic. Might it be good enough for at least localizing nuclei? (I do know how to do non-confocal UV imaging on the system) Can people advise on an NIR-to-UV swap of lasers?
Thanks,
Esteban
-- G. Esteban Fernandez, Ph.D. Associate Director Molecular Cytology Core Facility University of Missouri 120 Bond Life Sciences Center Columbia, MO 65211 http://www.biotech.missouri.edu/mcc/ 573-882-4895 573-884-9395 fax |
|
|
Hi Esteban,
Just a few thoughts, because I've never attempted such a swap of lasers: for DAPI, we use a 405 nm laser, and we found no need for real UV. However, even with this near-UV line, the light path is separate from the visible light paths and the near-UV optics contains correction pinhole lenses (one pinhole lens for each objective lens) in order to be able to align the visible and near-UV generated confocal images (even a slight misalignment of these corrective elements results in a 1 to 5 micron misalignment between the two images). We also image nuclei regularly and the light paths have to be well aligned if we want to have images of any use, thus I very much doubt that you would get away with a simple swap. I hope this helps a bit, I'm sure our more experienced list members will fill us in with great info soon. Best regards, Zoltan On Tue, Dec 23, 2008 at 4:47 PM, G. Esteban Fernandez <[hidden email]> wrote:
-- Zoltan Cseresnyes Facility manager, Imaging Suite |
|
|
Boswell, Carl A - (cboswell) |
Re: Changing system from NIR (two-photon) to UV
|
|
Quoting Zoltan Cseresnyes <[hidden email]>:
> Hi Esteban, > > Just a few thoughts, because I've never attempted such a swap of lasers: for > DAPI, we use a 405 nm laser, and we found no need for real UV. However, > even with this near-UV line, the light path is separate from the visible > light paths and the near-UV optics contains correction pinhole lenses (one > pinhole lens for each objective lens) in order to be able to align the > visible and near-UV generated confocal images (even a slight misalignment of > these corrective elements results in a 1 to 5 micron misalignment between > the two images). We also image nuclei regularly and the light paths have to > be well aligned if we want to have images of any use, thus I very much doubt > that you would get away with a simple swap. > I hope this helps a bit, I'm sure our more experienced list members will > fill us in with great info soon. Best regards, > > Zoltan > > On Tue, Dec 23, 2008 at 4:47 PM, G. Esteban Fernandez < > [hidden email]> wrote: > >> Hello all, >> >> On our Zeiss LSM 510 META two-photon system we have a Coherent Chameleon >> NIR laser that may need major repair. Coherent has been very good in >> providing remote technical assistance to us but we're at a point where a >> service visit is required and we're not under contract. Since we do more >> shallow imaging of UV dyes like DAPI on that system that two-photon imaging, >> we would replace the NIR laser with UV if it is more cost-effective than >> repairing the NIR. Zeiss says a switch from NIR to UV would require a major >> (and pricey) overhaul of the optics. I don't know what that entails but >> it is understandable given the huge difference in wavelength. However, I >> wonder how badly out of focus or aberrated the UV image would actually be >> with the optics that are already in place if we put a UV laser in there in >> place of the NIR; we do have a UV laser dichroic. Might it be good enough >> for at least localizing nuclei? (I do know how to do non-confocal UV imaging >> on the system) Can people advise on an NIR-to-UV swap of lasers? >> >> Thanks, >> Esteban >> >> -- >> G. Esteban Fernandez, Ph.D. >> Associate Director >> Molecular Cytology Core Facility >> University of Missouri >> 120 Bond Life Sciences Center >> Columbia, MO 65211 >> >> http://www.biotech.missouri.edu/mcc/ >> >> 573-882-4895 >> 573-884-9395 fax >> > > > > -- > > Zoltan Cseresnyes > Facility manager, Imaging Suite Hi Esteban, It is my understanding that the requirement for UV optics is from the excitation source to the microscope port. After that, you should be good to go and your objectives should be fine for the imaging. "Normal" fiber optics won't work for the 405 laser. In addition, collimators or other lenses in this path need to be UV capable. If this was not the focus of your question, my apologies. Carl |
|
|
Rosemary.White |
Re: Changing system from NIR (two-photon) to UV
|
|
In reply to this post by G. Esteban Fernandez
If you’re going to real UV, rather than just 405 nm, you need UV-transmitting optics, which means all of the optics in the light path, including the objectives, need to be replaced, and this really is quite expensive. The instrument usually has to go back to the factory, too. We added 405 nm laser to our confocal, which was much less expensive and could be done on site (was done in a day, tested and calibrated the next day), but still needed some change in scan head optics because this laser (in the Leica SP2 system) has a separate light path. The short answer is that it’s not straightforward and a major service visit and repair could well be cheaper than installing even the 405 nm laser. good luck, cheers, Rosemary Rosemary White CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia ph 61 2 6246 5475 fx 61 2 6246 5334 On 24/12/08 3:47 AM, "G. Esteban Fernandez" <g.esteban.fernandez@...> wrote: Hello all, |
|
|
Steve Ruzin |
Re: Changing system from NIR (two-photon) to UV
|
|
Hello Esteban:
I have a LSM 510 with a 365 UV laser (water cooled Ar laser). It's coupled to the scan head via a quartz fiber. Inside the head is a (factory) UV collimator lens. The objectives are all planNeo or Plan Apo. We have no "UV" (Ultrafluor) lenses. The cost is considerable: The laser is a Coherent Enterprise II system with a water-to-water heat exchanger. The laser was >$60k, and is now obsolete. I have a warranty directly from Coherent. To change your non-UV scan head would require sending back to the factory (Germany, not the US) to replace the primary dichroic, add a second fiber coupler, add the collimator, add the UV AOTF (factory-installed at the output window of the Coherent), and whatever other internal optics CZ requires. Given that the Enterprise is obsolete, you'll probably have to go with a UV diode, but they're not cheap either, and you still have the fiber and the AOTF to contend with. Steve... > > >On 24/12/08 3:47 AM, "G. Esteban Fernandez" ><<>[hidden email]> wrote: > >Hello all, > >On our Zeiss LSM 510 META two-photon system we have a Coherent >Chameleon NIR laser that may need major repair. Coherent has been >very good in providing remote technical assistance to us but we're >at a point where a service visit is required and we're not under >contract. Since we do more shallow imaging of UV dyes like DAPI on >that system that two-photon imaging, we would replace the NIR laser >with UV if it is more cost-effective than repairing the NIR. Zeiss >says a switch from NIR to UV would require a major (and pricey) >overhaul of the optics. I don't know what that entails but it is >understandable given the huge difference in wavelength. However, I >wonder how badly out of focus or aberrated the UV image would >actually be with the optics that are already in place if we put a UV >laser in there in place of the NIR; we do have a UV laser dichroic. > Might it be good enough for at least localizing nuclei? (I do know >how to do non-confocal UV imaging on the system) Can people advise >on an NIR-to-UV swap of lasers? > >Thanks, >Esteban -- ____________________________________________________________________________ Steven E. Ruzin, Ph.D. Director, Biological Imaging Facility 381 Koshland Hall College of Natural Resources University of California Berkeley CA 94720-3102 510-642-6602 510-642-4995 (fax) http://microscopy.berkeley.edu |
|
|
Holly L. AARON |
Re: Changing system from NIR (two-photon) to UV
|
|
Hi, Esteban - Echoing Steve's and others' comments, I would strongly advise
you to NOT send your scanhead back to Germany for an IR to UV change. It will be costly and you will be without your scanhead AND microscope during that time. At least, that's how we had to do it. We did not swap an IR for a UV, but added 2 VIS lasers and this required considerable time (and $$) in Germany. Zeiss considers the 405 diode to be a UV source, so it would definitely require a factory round-trip. Of course, it is possible if that is really what you want to do. Can you ask Coherent for an estimated price for service visit and replacement of diodes? Usually the only thing that can really go wrong on these lasers is the end of life on the diodes, or maybe some cleaning if you have really dusty labs like we have here. Also, the IR laser may be used primarily for DAPI now, but in the future has more potential uses... I would repair it. I'd be happy to listen to the symptoms of your laser and take a guess at the cause, if you want. I am sure the rest of the list could give you some good ideas on what is ailing, too. Best for 2009, -Holly Holly L. Aaron CRL Molecular Imaging Center [hidden email] The 6th Annual Advanced Imaging Methods Workshop is This January! Click the link below for more info: http://guest.cvent.com/EVENTS/Info/Summary.aspx?i=efa9c51b-9eba-4da5-a7e9-cd 0253f52cd3 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Steven Ruzin Sent: Tuesday, December 23, 2008 2:10 PM To: [hidden email] Subject: Re: Changing system from NIR (two-photon) to UV Hello Esteban: I have a LSM 510 with a 365 UV laser (water cooled Ar laser). It's coupled to the scan head via a quartz fiber. Inside the head is a (factory) UV collimator lens. The objectives are all planNeo or Plan Apo. We have no "UV" (Ultrafluor) lenses. The cost is considerable: The laser is a Coherent Enterprise II system with a water-to-water heat exchanger. The laser was >$60k, and is now obsolete. I have a warranty directly from Coherent. To change your non-UV scan head would require sending back to the factory (Germany, not the US) to replace the primary dichroic, add a second fiber coupler, add the collimator, add the UV AOTF (factory-installed at the output window of the Coherent), and whatever other internal optics CZ requires. Given that the Enterprise is obsolete, you'll probably have to go with a UV diode, but they're not cheap either, and you still have the fiber and the AOTF to contend with. Steve... > > >On 24/12/08 3:47 AM, "G. Esteban Fernandez" ><<>[hidden email]> wrote: > >Hello all, > >On our Zeiss LSM 510 META two-photon system we have a Coherent >Chameleon NIR laser that may need major repair. Coherent has been >very good in providing remote technical assistance to us but we're >at a point where a service visit is required and we're not under >contract. Since we do more shallow imaging of UV dyes like DAPI on >that system that two-photon imaging, we would replace the NIR laser >with UV if it is more cost-effective than repairing the NIR. Zeiss >says a switch from NIR to UV would require a major (and pricey) >overhaul of the optics. I don't know what that entails but it is >understandable given the huge difference in wavelength. However, I >wonder how badly out of focus or aberrated the UV image would >actually be with the optics that are already in place if we put a UV >laser in there in place of the NIR; we do have a UV laser dichroic. > Might it be good enough for at least localizing nuclei? (I do know >how to do non-confocal UV imaging on the system) Can people advise >on an NIR-to-UV swap of lasers? > >Thanks, >Esteban -- ____________________________________________________________________________ Steven E. Ruzin, Ph.D. Director, Biological Imaging Facility 381 Koshland Hall College of Natural Resources University of California Berkeley CA 94720-3102 510-642-6602 510-642-4995 (fax) http://microscopy.berkeley.edu |
|
|
R. Eric King |
Re: Changing system from NIR (two-photon) to UV
|
|
In reply to this post by Steve Ruzin
Hello Steve & List Members,
Pardon my commercial intrusion, but
---
Coherent may have discontinued the 651
& 653 UV models of the Enterprise I and II series
water cooled Ion laser systems, but we have
not.
Laser Innovations has plenty of these and many
other Coherent laser systems, plasma tubes,
and parts to service all OEM and end-user
customers, alike.
These Enterprise II UV laser systems retail from us
complete with the Coherent LP-5i "Water-to-Air"
heat exchanger for much lower than originally from
Coherent (<$30K), with the same
warranty.
Please let me know if you have any
questions, or require any further information. Thank you for your time,
and Best Regards,
R. Eric
King
LASER INNOVATIONS
www.CoherentLaser.com ----- Original Message -----
From: "Steven Ruzin" <[hidden email]>
To: <[hidden email]>
Sent: Tuesday, December 23, 2008 2:10
PM
Subject: Re: Changing system from NIR (two-photon)
to UV > > I have a LSM 510 with a 365 UV laser (water cooled Ar laser). It's > coupled to the scan head via a quartz fiber. Inside the head is a > (factory) UV collimator lens. The objectives are all planNeo or Plan > Apo. We have no "UV" (Ultrafluor) lenses. The cost is considerable: > The laser is a Coherent Enterprise II system with a water-to-water > heat exchanger. The laser was >$60k, and is now obsolete. I have a > warranty directly from Coherent. To change your non-UV scan head > would require sending back to the factory (Germany, not the US) to > replace the primary dichroic, add a second fiber coupler, add the > collimator, add the UV AOTF (factory-installed at the output window > of the Coherent), and whatever other internal optics CZ requires. > Given that the Enterprise is obsolete, you'll probably have to go > with a UV diode, but they're not cheap either, and you still have the > fiber and the AOTF to contend with. > > Steve... >> >> >>On 24/12/08 3:47 AM, "G. Esteban Fernandez" >><<>[hidden email]> wrote: >> >>Hello all, >> >>On our Zeiss LSM 510 META two-photon system we have a Coherent >>Chameleon NIR laser that may need major repair. Coherent has been >>very good in providing remote technical assistance to us but we're >>at a point where a service visit is required and we're not under >>contract. Since we do more shallow imaging of UV dyes like DAPI on >>that system that two-photon imaging, we would replace the NIR laser >>with UV if it is more cost-effective than repairing the NIR. Zeiss >>says a switch from NIR to UV would require a major (and pricey) >>overhaul of the optics. I don't know what that entails but it is >>understandable given the huge difference in wavelength. However, I >>wonder how badly out of focus or aberrated the UV image would >>actually be with the optics that are already in place if we put a UV >>laser in there in place of the NIR; we do have a UV laser dichroic. >> Might it be good enough for at least localizing nuclei? (I do know >>how to do non-confocal UV imaging on the system) Can people advise >>on an NIR-to-UV swap of lasers? >> >>Thanks, >>Esteban > > > -- > ____________________________________________________________________________ > Steven E. Ruzin, Ph.D. > Director, Biological Imaging Facility > 381 Koshland Hall College of Natural Resources > University of California Berkeley CA 94720-3102 > 510-642-6602 510-642-4995 > (fax) > http://microscopy.berkeley.edu |
|
|
Arvydas Matiukas |
Optimal LSM710 configuration
|
|
In reply to this post by G. Esteban Fernandez
Hello all,
we are considering buying Zeiss LSM710, most probably even LSM 7 DUO confocal system.
Current users/potential buyers of the LSM 7 family confocal microscopes please share your
experience and advice on optimal excitation, laser, scanning system, detector, and software
configuration.
We want to detemine some universal configuration that could serve most basic biomedical
imaging needs at medical university, and wouldn't require major upgrades for 5-7 years.
Thanks,
Arvydas
Director of Confocal&Two-Photon Imaging Core Facility Department of Pharmacology SUNY Upstate Medical University 766 Irving Ave., WH 3159 Syracuse, NY 13210 tel.: 315-464-7997 fax: 315-464-8014 email: [hidden email] |
«
Return to Confocal Microscopy List
|
1 view|%1 views
| Free forum by Nabble | Edit this page |