Olympus vs Nikon
Locked
|
This post was updated on .
CONTENTS DELETED
The author has deleted this message.
|
|
|
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****
Dear All,
____
Maria Mukhina
Postdoctoral Fellow Kleckner Lab Department of Molecular and Cellular Biology Harvard University
|
|
|
In reply to this post by muxika
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****
Hi Maria, this is going to be very objective lens dependent. Most
commercial "through objective" TIRF (o-TIRF) systems do not have
UV (such as 355 nm) lasers, so pretty much moot. Three
suggestions: * I suggest you meet with Jennifer Waters, Director of your
(Harvard's) Nikon Imaging Center, to discuss your needs, http://nic.med.harvard.edu/contact * You might also want to re-think about whether you even need UV ... STORM is much higher spatial precision localization than can be achieved with TIRF optical resolution -- Dr. STORM, aka Prof. Xiaowei Zhuang, is at Harvard, http://zhuang.harvard.edu * If you really need UV TIRF, I encourage you to discuss this directly with Dr. Alex Asanov, who I am copying, see http://www.tirf-labs.com/tirfmicroscopy.html
A big advantage of Alex's approach's (prism or light guide) is
you do not need a $20K TIRF objective lens. This would save you a
lot of money, since UV transmitting (assuming you really need to
collect UV emission) is likely a lot less expensive (I suspect
TIRF excitation lenses have even more glass than standard high NA
objective lenses, every lens is likely 2% or more decrease in
transmission ... see also QE curves of conventional sCMOS and
back-illuminated sCMOS, EMCCDs, CCDs for UV performance). The
money you save by going with either "Alex TIRF" approaches, could
also help pay for the UV laser(s) and optionally an LDI laser rack
-- see preliminary doc at
https://www.cairn-research.co.uk/wp-content/uploads/2016/03/A4-Leaflet-2.pdf
... http://www.89north.com would be selling the U.S. version (plus
would want appropriate emission filters). best wishes, George p.s. if you get this working, please let me know how well BUV395 and BV421 monoclonal antibodies work on it (Brilliant Ultraviolet and Brilliant Violet - see BD Biosciences).
On 12/22/2016 8:36 PM, Maria Mukhina
wrote:
***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi All, I wonder if anyone can share any experience in regard to relative transmission efficiency/optics threshold (especially in 300-400 nm UV)/software/automatic focus systems/TIRF modules of Olympus and Nikon microscopes. We are considering using microscopes of these two manufacturers as a base for custom setup and would appreciate any comments you might have. thanks in advance, Maria Maria Mukhina Postdoctoral Fellow Kleckner Lab Department of Molecular and Cellular Biology Harvard University -- George McNamara, PhD Houston, TX 77054 [hidden email] https://www.linkedin.com/in/georgemcnamara https://works.bepress.com/gmcnamara/75/ http://www.ncbi.nlm.nih.gov/myncbi/browse/collection/44962650 |
 
|
0000001ed7f52e4a-dmarc-request |
Re: Olympus vs Nikon
|
|
In reply to this post by muxika
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****
Dear Maria, Olympus has the 4 laser TIRF module where you can adjust the collimation and TIRF angle individually for each laser line. I think this is unique and it allows fast imaging in multi colours when combined with a camera splitter or even fast filter wheel. They publish the transmission curves of the objectives. I have no experience with Nikon. Best wishes Andreas Dear All,
____
Maria Mukhina
Postdoctoral Fellow Kleckner Lab Department of Molecular and Cellular Biology Harvard University
|
|
|
In reply to this post by muxika
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****
There are couple of high transmission UV optics with Olympus dedicated for Fura imaging. You may not good TIRF because of lower NA. Regards Ganesh On Dec 23, 2016 8:19 AM, "Maria Mukhina" <[hidden email]> wrote: ***** |
|
|
In reply to this post by muxika
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****
Sorry to repost.. You have couple of cool motorised TIRF from Olympus and a motorised drift compensator called IX3-ZDC2 ... You can check the same in their websites. One of the TIRF is unique in terms providing 4 parallel laser lines (with many power options) helps you to set same depth for all colors with all lasers focused to the BFP of the objective. You can also find High NA (UpTo 1.7) and a high mag (150X) objectives if you are using 16microm pixel EMCCD. Reg Ganesh On Dec 23, 2016 8:19 AM, "Maria Mukhina" <[hidden email]> wrote: ***** |
|
|
Alex Asanov |
Re: Olympus vs Nikon Commercial response
|
|
In reply to this post by muxika
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****
Hi Maria,
We have compared several TIRF objectives using 638,
532, and 465 nm lines. In agreement with the literature we found that the
intensitity of stray light was 15% or more at the TIRF surface relative to the
intensity of the evanescent wave. Out of 13 objectives tested Nikon lenses
produced less stray light. However, it is not a conclusive
observation, since the number 13 is small. Make a note that the contamination
increases exponentially, since the EW decays
exponentially.
Just a few measurements we performed with 405
nm excitation to realize that the intensity of autofluorescence and
scatter were too high.
The good news is that prism- and lightguide-based geometries are well suited for TIRFing with UV excitation: http://tirf-labs.com/Select_TIRF_geometry_WP.pdf.
Best
regards, Alexander Asanov,
Ph.D. From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Mukhina, Maria Sent: Thursday, December 22, 2016 10:49 PM To: [hidden email] Subject: Olympus vs Nikon Dear All, ____ Maria Mukhina Postdoctoral Fellow Kleckner Lab Department
of Molecular and Cellular Biology Harvard
University
|
|
|
Alex Asanov |
Re: Olympus vs Nikon Commercial response
|
|
In reply to this post by muxika
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****
Hi Maria,
We have compared several TIRF objectives using 638,
532, and 465 nm lines. In agreement with the literature we found that the
intensitity of stray light was 15% or more at the TIRF surface relative to the
intensity of the evanescent wave. Out of 13 objectives tested Nikon lenses
produced less stray light. However, it is not a conclusive
observation, since the number 13 is small. Make a note that the contamination
increases exponentially, since the EW decays
exponentially.
Just a few measurements we performed with 405
nm excitation to realize that the intensity of autofluorescence and
scatter were too high.
The good news is that prism- and lightguide-based geometries are well suited for TIRFing with UV excitation: http://tirf-labs.com/Select_TIRF_geometry_WP.pdf.
Best
regards, Alexander Asanov,
Ph.D. From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Mukhina, Maria Sent: Thursday, December 22, 2016 10:49 PM To: [hidden email] Subject: Olympus vs Nikon Dear All, ____ Maria Mukhina Postdoctoral Fellow Kleckner Lab Department
of Molecular and Cellular Biology Harvard
University
|
|
|
In reply to this post by 0000001ed7f52e4a-dmarc-request
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi All, Thanks a lot for your replies. TIRF is not our priority in this setup. Mostly we are interested in maximal transmission efficiency and high optics threshold. Also we are quite flexible with excitation wavelength in UV. Though the best option is to excite at 300 nm, the excitation wavelength can be shifted to 330-350 nm, and emission wavelength is in the visible range. Since excitation at 300 nm requires very custom optics, we basically excluded this option and are considering using more conventional objectives. It looks like Olympus offers more options for UV imaging, and we are considering UAPON 100XOTIRF objective. In this regard, it would be very helpful to know its power threshold at UV range, especially in TIRF mode. Another important feature is automatic focus system. We currently use Nikon's Perfect Focus System and are satisfied with results. @Ganesh: Have you tried using Olympus drift compensator with living specimens? thanks, Maria ____ Maria Mukhina Postdoctoral Fellow Kleckner Lab Department of Molecular and Cellular Biology Harvard University ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Andreas Bruckbauer <[hidden email]> Sent: Friday, December 23, 2016 3:15:41 AM To: [hidden email] Subject: Re: Olympus vs Nikon ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy<https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=CwMFaQ&c=WO-RGvefibhHBZq3fL85hQ&r=lvsgf9vctNrfAxRUoAziiQY-eNA2rMSgY-3fOZjtYas&m=VQ87tNDZ4wzO9DfQjPsEPEyrALO87R2NN1VXVptSF3g&s=5u0Szuar1C82RB3RPtJ-M_0nroa8j1MQHVbCEPZ9QZ4&e=> Post images on http://www.imgur.com<https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=CwMFaQ&c=WO-RGvefibhHBZq3fL85hQ&r=lvsgf9vctNrfAxRUoAziiQY-eNA2rMSgY-3fOZjtYas&m=VQ87tNDZ4wzO9DfQjPsEPEyrALO87R2NN1VXVptSF3g&s=s8nje2bpAS463c91VsSOkQnoGn4gGCTZJrnY5YjU9tg&e=> and include the link in your posting. ***** Dear Maria, Olympus has the 4 laser TIRF module where you can adjust the collimation and TIRF angle individually for each laser line. I think this is unique and it allows fast imaging in multi colours when combined with a camera splitter or even fast filter wheel. They publish the transmission curves of the objectives. I have no experience with Nikon. Best wishes Andreas ________________________________ From: Mukhina, Maria<mailto:[hidden email]> Sent: ?23/?12/?2016 02:49 To: [hidden email]<mailto:[hidden email]> Subject: Olympus vs Nikon ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy<https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=CwMFaQ&c=WO-RGvefibhHBZq3fL85hQ&r=lvsgf9vctNrfAxRUoAziiQY-eNA2rMSgY-3fOZjtYas&m=VQ87tNDZ4wzO9DfQjPsEPEyrALO87R2NN1VXVptSF3g&s=5u0Szuar1C82RB3RPtJ-M_0nroa8j1MQHVbCEPZ9QZ4&e=> Post images on http://www.imgur.com<https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=CwMFaQ&c=WO-RGvefibhHBZq3fL85hQ&r=lvsgf9vctNrfAxRUoAziiQY-eNA2rMSgY-3fOZjtYas&m=VQ87tNDZ4wzO9DfQjPsEPEyrALO87R2NN1VXVptSF3g&s=s8nje2bpAS463c91VsSOkQnoGn4gGCTZJrnY5YjU9tg&e=> and include the link in your posting. ***** Dear All, I wonder if anyone can share any experience in regard to relative transmission efficiency/optics threshold (especially in 300-400 nm UV)/software/automatic focus systems/TIRF modules of Olympus and Nikon microscopes. We are considering using microscopes of these two manufacturers as a base for custom setup and would appreciate any comments you might have. thanks in advance, Maria ____ Maria Mukhina Postdoctoral Fellow Kleckner Lab Department of Molecular and Cellular Biology Harvard University |
|
|
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Hi All, @Mike Could you please tell what source you use at 340 nm? @Ganesh Thank you. Happy New Year! Maria ____ Maria Mukhina Postdoctoral Fellow Kleckner Lab Department of Molecular and Cellular Biology Harvard University ________________________________ From: Confocal Microscopy List <[hidden email]> on behalf of Ganesh Kadasoor <[hidden email]> Sent: Wednesday, December 28, 2016 1:33:31 AM To: [hidden email] Subject: Re: Olympus vs Nikon ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy Post images on http://www.imgur.com and include the link in your posting. ***** Dear Maria I have used the n IX3-ZDC which goes with IX83 inverted scope. It works fine both in single shot and continuous mode. I have also seen people using it more than 24 hrs without any issues. Regards Ganesh Sent from my iPhone > On 28-Dec-2016, at 12:37 AM, MODEL, MICHAEL <[hidden email]> wrote: > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images on http://www.imgur.com and include the link in your posting. > ***** > > Hi Maria, > > We use Olympus UPlanSApo 20/0.75 (or an old 40x oil) for 340 excitation. At this wavelength, some slides can be very autofluorescent, so it's better to use an open coverslipped dish. > > Happy holidays! > > Mike > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Mukhina, Maria > Sent: Tuesday, December 27, 2016 1:58 PM > To: [hidden email] > Subject: Re: Olympus vs Nikon > > ***** > To join, leave or search the confocal microscopy listserv, go to: > https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=zQTnORo1bprZncYw%2BR3kw3LFLQYTiePWzqtO%2FWNMW6E%3D&reserved=0 > Post images on https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=yqQIkHZ2VbKnHDfSzeqc%2FXShoAstahva2OHqenbJW0I%3D&reserved=0 and include the link in your posting. > ***** > > Hi All, > > > Thanks a lot for your replies. TIRF is not our priority in this setup. Mostly we are interested in maximal transmission efficiency and high optics threshold. Also we are quite flexible with excitation wavelength in UV. Though the best option is to excite at 300 nm, the excitation wavelength can be shifted to 330-350 nm, and emission wavelength is in the visible range. Since excitation at 300 nm requires very custom optics, we basically excluded this option and are considering using more conventional objectives. It looks like Olympus offers more options for UV imaging, and we are considering UAPON 100XOTIRF objective. In this regard, it would be very helpful to know its power threshold at UV range, especially in TIRF mode. > > > Another important feature is automatic focus system. We currently use Nikon's Perfect Focus System and are satisfied with results. @Ganesh: Have you tried using Olympus drift compensator with living specimens? > > > thanks, > > Maria > > > ____ > > > Maria Mukhina > > > Postdoctoral Fellow > > Kleckner Lab > > Department of Molecular and Cellular Biology > > Harvard University > > > ________________________________ > From: Confocal Microscopy List <[hidden email]> on behalf of Andreas Bruckbauer <[hidden email]> > Sent: Friday, December 23, 2016 3:15:41 AM > To: [hidden email] > Subject: Re: Olympus vs Nikon > > ***** To join, leave or search the confocal microscopy listserv, go to: https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=zQTnORo1bprZncYw%2BR3kw3LFLQYTiePWzqtO%2FWNMW6E%3D&reserved=0<https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.proofpoint.com%2Fv2%2Furl%3Fu%3Dhttp-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy%26d%3DCwMFaQ%26c%3DWO-RGvefibhHBZq3fL85hQ%26r%3Dlvsgf9vctNrfAxRUoAziiQY-eNA2rMSgY-3fOZjtYas%26m%3DVQ87tNDZ4wzO9DfQjPsEPEyrALO87R2NN1VXVptSF3g%26s%3D5u0Szuar1C82RB3RPtJ-M_0nroa8j1MQHVbCEPZ9QZ4%26e&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=YLKO3n7IMVOkyOobeT3DLnnVBOJX6xGdjeNvwQxsYSc%3D&reserved=0=> Post images on https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=yqQIkHZ2VbKnHDfSzeqc%2FXShoAstahva2OHqenbJW0I%3D&reserved=0<https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.proofpoint.com%2Fv2%2Furl%3Fu%3Dhttp-3A__www.imgur.com%26d%3DCwMFaQ%26c%3DWO-RGvefibhHBZq3fL85hQ%26r%3Dlvsgf9vctNrfAxRUoAziiQY-eNA2rMSgY-3fOZjtYas%26m%3DVQ87tNDZ4wzO9DfQjPsEPEyrALO87R2NN1VXVptSF3g%26s%3Ds8nje2bpAS463c91VsSOkQnoGn4gGCTZJrnY5YjU9tg%26e&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=i09q85%2FthtbPVgOqcWt3hsJo6vhByv04qkqxp53xRkE%3D&reserved=0=> and include the link in your posting. ***** Dear Maria, Olympus has the 4 laser TIRF module where you can adjust the collimation and TIRF angle individually for each laser line. I think this is unique and it allows fast imaging in multi colours when combined with a camera splitter or even fast filter wheel. They publish the transmission curves of the objectives. I have no experience with Nikon. > > Best wishes > > Andreas > > ________________________________ > From: Mukhina, Maria<mailto:[hidden email]> > Sent: ?23/?12/?2016 02:49 > To: [hidden email]<mailto:[hidden email]> > Subject: Olympus vs Nikon > > ***** To join, leave or search the confocal microscopy listserv, go to: https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=zQTnORo1bprZncYw%2BR3kw3LFLQYTiePWzqtO%2FWNMW6E%3D&reserved=0<https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.proofpoint.com%2Fv2%2Furl%3Fu%3Dhttp-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy%26d%3DCwMFaQ%26c%3DWO-RGvefibhHBZq3fL85hQ%26r%3Dlvsgf9vctNrfAxRUoAziiQY-eNA2rMSgY-3fOZjtYas%26m%3DVQ87tNDZ4wzO9DfQjPsEPEyrALO87R2NN1VXVptSF3g%26s%3D5u0Szuar1C82RB3RPtJ-M_0nroa8j1MQHVbCEPZ9QZ4%26e&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=YLKO3n7IMVOkyOobeT3DLnnVBOJX6xGdjeNvwQxsYSc%3D&reserved=0=> Post images on https://na01.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=yqQIkHZ2VbKnHDfSzeqc%2FXShoAstahva2OHqenbJW0I%3D&reserved=0<https://na01.safelinks.protection.outlook.com/?url=https%3A%2F%2Furldefense.proofpoint.com%2Fv2%2Furl%3Fu%3Dhttp-3A__www.imgur.com%26d%3DCwMFaQ%26c%3DWO-RGvefibhHBZq3fL85hQ%26r%3Dlvsgf9vctNrfAxRUoAziiQY-eNA2rMSgY-3fOZjtYas%26m%3DVQ87tNDZ4wzO9DfQjPsEPEyrALO87R2NN1VXVptSF3g%26s%3Ds8nje2bpAS463c91VsSOkQnoGn4gGCTZJrnY5YjU9tg%26e&data=01%7C01%7Cmmodel%40KENT.EDU%7C55f9a4e29d4942e5052508d42e8a5179%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1&sdata=i09q85%2FthtbPVgOqcWt3hsJo6vhByv04qkqxp53xRkE%3D&reserved=0=> and include the link in your posting. ***** > > Dear All, > > I wonder if anyone can share any experience in regard to relative transmission efficiency/optics threshold (especially in 300-400 nm UV)/software/automatic focus systems/TIRF modules of Olympus and Nikon microscopes. We are considering using microscopes of these two manufacturers as a base for custom setup and would appreciate any comments you might have. > > thanks in advance, > Maria > > > > ____ > > > Maria Mukhina > > > Postdoctoral Fellow > > Kleckner Lab > > Department of Molecular and Cellular Biology > > Harvard University |
«
Return to Confocal Microscopy List
|
1 view|%1 views
| Free forum by Nabble | Edit this page |