Zeiss spinning disc confocal microscope
Locked
 
|
Julia Edgar |
Zeiss spinning disc confocal microscope
|
*****
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 Can anybody tell me how to adjust the camera focus so that it synchronises with the focal plane we observe by eye? At present we have to adjust the focal plane after switching to camera. I don't believe we should have to do this. Many thanks Best wishes Julia Julia M Edgar BSc(Hons), PhD, FHEA Institute of Infection, Immunity and Inflammation College of Medical Veterinary and Life Sciences University of Glasgow Sir Graeme Davies Building 120 University Place Glasgow G12 8TA UK Tel 0141 330 2082 https://www.facebook.com/Neuroimmunology-group-at-the-University-of-Glasgow-1741018122797881/ www.facebook.com/iiiiglasgow And Department of Neurogenetics Max Planck Institute for Experimental Medicine Hermann-Rein-Strasse 3 D-37075 Goettingen Germany |
|
|
George McNamara |
Re: Zeiss spinning disc confocal microscope
|
|
*****
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 Julia, Best to adjust the EYEPIECES to match the camera. Most research microscopes have adjustable eyepieces. Typically these have a "tick mark" (or ring) to mark the eyepiece focus for 20/20 vision. Best to start with a medium to long working distance objective lens. Procedure: Get close to focus. Set both eyepieces to "20/20" setting. Use the camera to focus (if far from focus, be careful not to smash anything). Look in the eyepieces - should match for users with 20/20 vision. IF NOT - could be the user(s) do not have 20/20 vision and/or the way the camera is mounted on the microscope is "way off" (there is not much that can go wrong on a simple 1x C-mount adapter). Personally, I rarely used the eyepieces. Many visitors to my former microscope room at MDACC saw my (still) patent pending cover for the Leica DMI6000 microscope to keep light from entering the eyepieces and reaching the ORCA FLASH 4.2 sCMOS. I rarely removed the cover (not that I paid myself royalties to do so). NOTE: some objective lenses have "correction collars" for coverglass thickness or refractive index. Adjust eyepiece focus (and if needed deal with any weird camera mount issues) with a standard dry objective lens (i.e. 20x/0.8NA lens, no correction collar), then switch objective lenses, correct the correction collar to match the standard lens. enjoy, George p.s. hopefully the spinning disk component does not severely mess with the parfocality, or have some secret parfocality adjustment that is far from adjustment). On 10/18/2016 1:11 PM, Julia Edgar 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 > Can anybody tell me how to adjust the camera focus so that it synchronises with the focal plane we observe by eye? > At present we have to adjust the focal plane after switching to camera. I don't believe we should have to do this. > Many thanks > Best wishes > Julia > > Julia M Edgar BSc(Hons), PhD, FHEA > Institute of Infection, Immunity and Inflammation > College of Medical Veterinary and Life Sciences > University of Glasgow > Sir Graeme Davies Building > 120 University Place > Glasgow > G12 8TA > UK > Tel 0141 330 2082 > https://www.facebook.com/Neuroimmunology-group-at-the-University-of-Glasgow-1741018122797881/ > > www.facebook.com/iiiiglasgow > > And > > Department of Neurogenetics > Max Planck Institute for Experimental Medicine > Hermann-Rein-Strasse 3 > D-37075 Goettingen > Germany -- 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 |
|
|
John Oreopoulos |
Re: Zeiss spinning disc confocal microscope
|
|
*****
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. ***** If the adjustment is beyond the range of the eyepieces as George suggests, then you might need to change the axial position of the confocal scan head relative to the microscope side port. You'll also get a more symmetric PSF and better axial sectioning when the pinhole disk is matched to the tube lens focus. If your scan head is hard mounted to the microscope side port, you should talk to your Zeiss rep. It might need to be adjusted, or maybe you've got the wrong mount on there. John Oreopoulos Quoting George McNamara <[hidden email]>: > ***** > 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 Julia, > > Best to adjust the EYEPIECES to match the camera. > > Most research microscopes have adjustable eyepieces. Typically these > have a "tick mark" (or ring) to mark the eyepiece focus for 20/20 > vision. Best to start with a medium to long working distance objective > lens. > > Procedure: > > Get close to focus. > > Set both eyepieces to "20/20" setting. > > Use the camera to focus (if far from focus, be careful not to smash > anything). > > Look in the eyepieces - should match for users with 20/20 vision. IF > NOT - could be the user(s) do not have 20/20 vision and/or the way the > camera is mounted on the microscope is "way off" (there is not much > that can go wrong on a simple 1x C-mount adapter). > > Personally, I rarely used the eyepieces. Many visitors to my former > microscope room at MDACC saw my (still) patent pending cover for the > Leica DMI6000 microscope to keep light from entering the eyepieces and > reaching the ORCA FLASH 4.2 sCMOS. I rarely removed the cover (not that > I paid myself royalties to do so). > > NOTE: some objective lenses have "correction collars" for coverglass > thickness or refractive index. Adjust eyepiece focus (and if needed > deal with any weird camera mount issues) with a standard dry objective > lens (i.e. 20x/0.8NA lens, no correction collar), then switch objective > lenses, correct the correction collar to match the standard lens. > > enjoy, > > George > > p.s. hopefully the spinning disk component does not severely mess with > the parfocality, or have some secret parfocality adjustment that is far > from adjustment). > > > On 10/18/2016 1:11 PM, Julia Edgar 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 >> Can anybody tell me how to adjust the camera focus so that it >> synchronises with the focal plane we observe by eye? >> At present we have to adjust the focal plane after switching to >> camera. I don't believe we should have to do this. >> Many thanks >> Best wishes >> Julia >> >> Julia M Edgar BSc(Hons), PhD, FHEA >> Institute of Infection, Immunity and Inflammation >> College of Medical Veterinary and Life Sciences >> University of Glasgow >> Sir Graeme Davies Building >> 120 University Place >> Glasgow >> G12 8TA >> UK >> Tel 0141 330 2082 >> https://www.facebook.com/Neuroimmunology-group-at-the-University-of-Glasgow-1741018122797881/ >> >> www.facebook.com/iiiiglasgow >> >> And >> >> Department of Neurogenetics >> Max Planck Institute for Experimental Medicine >> Hermann-Rein-Strasse 3 >> D-37075 Goettingen >> Germany > > -- > > > 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 |
|
|
Julia Edgar |
Re: Zeiss spinning disc confocal microscope
|
|
In reply to this post by George McNamara
*****
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. ***** Many thanks George That's what I thought I should do, but this one doesn't have adjustable eyepieces (as far as I can tell). Best wishes Julia -----Original Message----- From: George McNamara [mailto:[hidden email]] Sent: 18 October 2016 19:29 To: Confocal Microscopy List; Julia Edgar Subject: Re: Zeiss spinning disc confocal microscope Hi Julia, Best to adjust the EYEPIECES to match the camera. Most research microscopes have adjustable eyepieces. Typically these have a "tick mark" (or ring) to mark the eyepiece focus for 20/20 vision. Best to start with a medium to long working distance objective lens. Procedure: Get close to focus. Set both eyepieces to "20/20" setting. Use the camera to focus (if far from focus, be careful not to smash anything). Look in the eyepieces - should match for users with 20/20 vision. IF NOT - could be the user(s) do not have 20/20 vision and/or the way the camera is mounted on the microscope is "way off" (there is not much that can go wrong on a simple 1x C-mount adapter). Personally, I rarely used the eyepieces. Many visitors to my former microscope room at MDACC saw my (still) patent pending cover for the Leica DMI6000 microscope to keep light from entering the eyepieces and reaching the ORCA FLASH 4.2 sCMOS. I rarely removed the cover (not that I paid myself royalties to do so). NOTE: some objective lenses have "correction collars" for coverglass thickness or refractive index. Adjust eyepiece focus (and if needed deal with any weird camera mount issues) with a standard dry objective lens (i.e. 20x/0.8NA lens, no correction collar), then switch objective lenses, correct the correction collar to match the standard lens. enjoy, George p.s. hopefully the spinning disk component does not severely mess with the parfocality, or have some secret parfocality adjustment that is far from adjustment). On 10/18/2016 1:11 PM, Julia Edgar 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 > Can anybody tell me how to adjust the camera focus so that it synchronises with the focal plane we observe by eye? > At present we have to adjust the focal plane after switching to camera. I don't believe we should have to do this. > Many thanks > Best wishes > Julia > > Julia M Edgar BSc(Hons), PhD, FHEA > Institute of Infection, Immunity and Inflammation College of Medical > Veterinary and Life Sciences University of Glasgow Sir Graeme Davies > Building > 120 University Place > Glasgow > G12 8TA > UK > Tel 0141 330 2082 > https://www.facebook.com/Neuroimmunology-group-at-the-University-of-Gl > asgow-1741018122797881/ > > www.facebook.com/iiiiglasgow > > And > > Department of Neurogenetics > Max Planck Institute for Experimental Medicine Hermann-Rein-Strasse 3 > D-37075 Goettingen > Germany -- 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 |
|
|
Gary Laevsky |
Re: Zeiss spinning disc confocal microscope
|
|
*****
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. ***** Depending on the configuration of the c-mount, you might even be able to pull it out a few mm from the stand. Best, Gary > On Oct 18, 2016, at 3:19 PM, Julia Edgar <[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. > ***** > > Many thanks George > That's what I thought I should do, but this one doesn't have adjustable eyepieces (as far as I can tell). > Best wishes > Julia > > -----Original Message----- > From: George McNamara [mailto:[hidden email]] > Sent: 18 October 2016 19:29 > To: Confocal Microscopy List; Julia Edgar > Subject: Re: Zeiss spinning disc confocal microscope > > Hi Julia, > > Best to adjust the EYEPIECES to match the camera. > > Most research microscopes have adjustable eyepieces. Typically these have a "tick mark" (or ring) to mark the eyepiece focus for 20/20 vision. Best to start with a medium to long working distance objective lens. > > Procedure: > > Get close to focus. > > Set both eyepieces to "20/20" setting. > > Use the camera to focus (if far from focus, be careful not to smash anything). > > Look in the eyepieces - should match for users with 20/20 vision. IF NOT > - could be the user(s) do not have 20/20 vision and/or the way the camera is mounted on the microscope is "way off" (there is not much that can go wrong on a simple 1x C-mount adapter). > > Personally, I rarely used the eyepieces. Many visitors to my former microscope room at MDACC saw my (still) patent pending cover for the Leica DMI6000 microscope to keep light from entering the eyepieces and reaching the ORCA FLASH 4.2 sCMOS. I rarely removed the cover (not that I paid myself royalties to do so). > > NOTE: some objective lenses have "correction collars" for coverglass thickness or refractive index. Adjust eyepiece focus (and if needed deal with any weird camera mount issues) with a standard dry objective lens (i.e. 20x/0.8NA lens, no correction collar), then switch objective lenses, correct the correction collar to match the standard lens. > > enjoy, > > George > > p.s. hopefully the spinning disk component does not severely mess with the parfocality, or have some secret parfocality adjustment that is far from adjustment). > > >> On 10/18/2016 1:11 PM, Julia Edgar 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 >> Can anybody tell me how to adjust the camera focus so that it synchronises with the focal plane we observe by eye? >> At present we have to adjust the focal plane after switching to camera. I don't believe we should have to do this. >> Many thanks >> Best wishes >> Julia >> >> Julia M Edgar BSc(Hons), PhD, FHEA >> Institute of Infection, Immunity and Inflammation College of Medical >> Veterinary and Life Sciences University of Glasgow Sir Graeme Davies >> Building >> 120 University Place >> Glasgow >> G12 8TA >> UK >> Tel 0141 330 2082 >> https://www.facebook.com/Neuroimmunology-group-at-the-University-of-Gl >> asgow-1741018122797881/ >> >> www.facebook.com/iiiiglasgow >> >> And >> >> Department of Neurogenetics >> Max Planck Institute for Experimental Medicine Hermann-Rein-Strasse 3 >> D-37075 Goettingen >> Germany > > -- > > > 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 |
|
|
Craig Brideau |
Re: Zeiss spinning disc confocal microscope
|
|
*****
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.
*****
The Nikon Ti frame with a C1 or C2 scanhead has a set of spacers on the tube going into the port. The spacers are selected based on the frame and set the scanhead at the correct distance. You might have a similar feature on the mount of your own scanhead. CraigOn Tue, Oct 18, 2016 at 4:34 PM, Gary Laevsky <[hidden email]> wrote: ***** |
«
Return to Confocal Microscopy List
|
1 view|%1 views
| Free forum by Nabble | Edit this page |