vertical stage travel
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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, Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. Mike |
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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. ***** Measure a coverslip -not forgetting about RI effects? HTH Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 3:46 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [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. ***** Dear all, Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. Mike |
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In reply to this post by mmodel
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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 a coverslip is placed on another at one side you've made an air wedge so that now lateral displacement can give small changes in Z position of the coverslip surface HTH Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 3:46 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [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. ***** Dear all, Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. Mike |
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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 Mike, If you can do TIRF on this setup maybe the fluorescently labelled beads of a uniform size would be helpful to roughly estimate Z axis distance. Shuce Shuce Zhang B.Sc., Ph.D. Candidate Department of Chemistry, University of Alberta Centennial Centre for Interdisciplinary Science 4-137 11227 Saskatchewan Drive, Edmonton AB T6G 2G2 Canada [hidden email] | www.ualberta.ca/~shuce > On Mar 4, 2021, at 8:52 AM, Mark Cannell <[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. > ***** > > If a coverslip is placed on another at one side you've made an air wedge so that now lateral displacement can give small changes in Z position of the coverslip surface > > HTH > > Mark B. Cannell. Ph.D. FRSNZ FISHR > Department of Physiology, Pharmacology & Neuroscience > School of Medical Sciences > University Walk > Bristol BS8 1TD > > [hidden email] > > > > On 4/03/21, 3:46 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [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. > ***** > > Dear all, > > Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. > > Mike > |
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In reply to this post by Shuce Zhang
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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. ***** We actually do have TIRF on that scope. Maybe something can be devised using TIRF. Less sure about beads because then you have two unknowns: the z step and the vertical PSF. On the other hand, if we image subresolution beads and assume that PSF is like theoretical, then maybe. Thanks for the idea! -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Shuce Zhang Sent: Thursday, March 4, 2021 11:00 AM To: [hidden email] Subject: Re: vertical stage travel ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7C263ba5bab36649a3676d08d8df280265%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504710228392935%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=lzzHc8siAQrncXLhHUyR9RQSfvN5b%2BowU1%2FSpRllZ9A%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C263ba5bab36649a3676d08d8df280265%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504710228392935%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=QgseNX3lwUiIL2Wto7rKSaxjSDUSvh6hh7JX7NCka4E%3D&reserved=0 and include the link in your posting. ***** Hi Mike, If you can do TIRF on this setup maybe the fluorescently labelled beads of a uniform size would be helpful to roughly estimate Z axis distance. Shuce Shuce Zhang B.Sc., Ph.D. Candidate Department of Chemistry, University of Alberta Centennial Centre for Interdisciplinary Science 4-137 11227 Saskatchewan Drive, Edmonton AB T6G 2G2 Canada [hidden email] | https://nam11.safelinks.protection.outlook.com/?url=http:%2F%2Fwww.ualberta.ca%2F~shuce&data=04%7C01%7Cmmodel%40KENT.EDU%7C263ba5bab36649a3676d08d8df280265%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504710228392935%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=whoxkIUAA%2FAJYFNszXV236R%2FXahdvKPsFPqjSwDrvW8%3D&reserved=0 > On Mar 4, 2021, at 8:52 AM, Mark Cannell <[hidden email]> wrote: > > ***** > To join, leave or search the confocal microscopy listserv, go to: > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists > .umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cm > model%40KENT.EDU%7C263ba5bab36649a3676d08d8df280265%7Ce5a06f4a1ec44d01 > 8f73e7dd15f26134%7C1%7C0%7C637504710228392935%7CUnknown%7CTWFpbGZsb3d8 > eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1 > 000&sdata=lzzHc8siAQrncXLhHUyR9RQSfvN5b%2BowU1%2FSpRllZ9A%3D&r > eserved=0 Post images on > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C263ba5bab36649a3676d08d8df280265%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504710228392935%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=QgseNX3lwUiIL2Wto7rKSaxjSDUSvh6hh7JX7NCka4E%3D&reserved=0 and include the link in your posting. > ***** > > If a coverslip is placed on another at one side you've made an air > wedge so that now lateral displacement can give small changes in Z > position of the coverslip surface > > HTH > > Mark B. Cannell. Ph.D. FRSNZ FISHR > Department of Physiology, Pharmacology & Neuroscience School of > Medical Sciences University Walk Bristol BS8 1TD > > [hidden email] > > > > On 4/03/21, 3:46 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [hidden email]> wrote: > > ***** > To join, leave or search the confocal microscopy listserv, go to: > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7C263ba5bab36649a3676d08d8df280265%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504710228392935%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=lzzHc8siAQrncXLhHUyR9RQSfvN5b%2BowU1%2FSpRllZ9A%3D&reserved=0 > Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C263ba5bab36649a3676d08d8df280265%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504710228392935%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=QgseNX3lwUiIL2Wto7rKSaxjSDUSvh6hh7JX7NCka4E%3D&reserved=0 and include the link in your posting. > ***** > > Dear all, > > Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. > > Mike > |
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In reply to this post by mmodel
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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. ***** Measure the thickness (csthick) of a coverslip and since that coverslip is placed to one side to create the wedge between the normal objective coverslip and a second test coverslip on top (width cswidth) you can use: dz=dx*sin(arctan(csheight/cswidth))) Where dz is the change in height to the test surface and dx is the lateral displacement. How you make the test coverslip surface visible is up to you -dirt, smoke particles, dried (fluorescent?) beads etc. If you want to get fancy you could even measure the change in height by observing the fringes in the wedge under monochromatic light ala Newton's fringe experiments __ HTH Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 4:41 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [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. ***** Thanks! But how would you determine the exact height of a coverslip within less then a micron on a widefield microscope? -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Cannell Sent: Thursday, March 4, 2021 10:52 AM To: [hidden email] Subject: Re: vertical stage travel ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2BogpChJQXFCwavjPUSymecSHTqnhzwdxqD9yfUhn7Q%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=i2y8SAPoXZ7KUAL6iuPZ%2F2WhXz4oB2kRtRmXZRThmZU%3D&reserved=0 and include the link in your posting. ***** If a coverslip is placed on another at one side you've made an air wedge so that now lateral displacement can give small changes in Z position of the coverslip surface HTH Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 3:46 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [hidden email]> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2BogpChJQXFCwavjPUSymecSHTqnhzwdxqD9yfUhn7Q%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=i2y8SAPoXZ7KUAL6iuPZ%2F2WhXz4oB2kRtRmXZRThmZU%3D&reserved=0 and include the link in your posting. ***** Dear all, Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. Mike |
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In reply to this post by mmodel
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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. ***** When doing these calculations, remember that each change in refractive index will change the perceived height. I forgot to do this a few years ago when measuring the depth of a microfabricated chamber for someone with reflection microscopy on a confocal. I was puzzled why the measurements were so far off. Correcting for RI gave the correct measurement. Cheers- Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY 10016 Office: 646-501-0567 Cell: 914-309-3270 [hidden email] http://nyulmc.org/micros http://microscopynotes.com/ -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of MODEL, MICHAEL Sent: Thursday, March 4, 2021 11:33 AM To: [hidden email] Subject: Re: vertical stage travel [EXTERNAL] ***** To join, leave or search the confocal microscopy listserv, go to: https://urldefense.com/v3/__http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy__;!!MXfaZl3l!J6dqBvIoDHd_olJ7YGD6cXS5EPW5EOrK2ziyUZsIX2NApail1DNiZ2Xvf_W5IwkaKt7oJUA$ Post images on https://urldefense.com/v3/__http://www.imgur.com__;!!MXfaZl3l!J6dqBvIoDHd_olJ7YGD6cXS5EPW5EOrK2ziyUZsIX2NApail1DNiZ2Xvf_W5Iwka6bxdk5w$ and include the link in your posting. ***** Thanks! But how would you determine the exact height of a coverslip within less then a micron on a widefield microscope? -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Cannell Sent: Thursday, March 4, 2021 10:52 AM To: [hidden email] Subject: Re: vertical stage travel ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2BogpChJQXFCwavjPUSymecSHTqnhzwdxqD9yfUhn7Q%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=i2y8SAPoXZ7KUAL6iuPZ%2F2WhXz4oB2kRtRmXZRThmZU%3D&reserved=0 and include the link in your posting. ***** If a coverslip is placed on another at one side you've made an air wedge so that now lateral displacement can give small changes in Z position of the coverslip surface HTH Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 3:46 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [hidden email]> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2BogpChJQXFCwavjPUSymecSHTqnhzwdxqD9yfUhn7Q%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=i2y8SAPoXZ7KUAL6iuPZ%2F2WhXz4oB2kRtRmXZRThmZU%3D&reserved=0 and include the link in your posting. ***** Dear all, Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. Mike |
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In reply to this post by Mark Cannell-2
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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. ***** As an example, for a wedge created by a coverslip 20 mm wide placed on a 160 um thick coverslip (measured with a micrometer) the height change will be 8 um per mm lateral movement of the wedge (actually don’t need trig can use similar triangles -silly me . This should be fine enough to measure your z stage accuracy (and backlash?) If the field curvature of an objective is not large you'll see particles on its surface over in and out of focus across the field .... Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 4:55 PM, "Confocal Microscopy List on behalf of Mark Cannell" <[hidden email] on behalf of [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. ***** Measure the thickness (csthick) of a coverslip and since that coverslip is placed to one side to create the wedge between the normal objective coverslip and a second test coverslip on top (width cswidth) you can use: dz=dx*sin(arctan(csheight/cswidth))) Where dz is the change in height to the test surface and dx is the lateral displacement. How you make the test coverslip surface visible is up to you -dirt, smoke particles, dried (fluorescent?) beads etc. If you want to get fancy you could even measure the change in height by observing the fringes in the wedge under monochromatic light ala Newton's fringe experiments __ HTH Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 4:41 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [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. ***** Thanks! But how would you determine the exact height of a coverslip within less then a micron on a widefield microscope? -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Cannell Sent: Thursday, March 4, 2021 10:52 AM To: [hidden email] Subject: Re: vertical stage travel ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2BogpChJQXFCwavjPUSymecSHTqnhzwdxqD9yfUhn7Q%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=i2y8SAPoXZ7KUAL6iuPZ%2F2WhXz4oB2kRtRmXZRThmZU%3D&reserved=0 and include the link in your posting. ***** If a coverslip is placed on another at one side you've made an air wedge so that now lateral displacement can give small changes in Z position of the coverslip surface HTH Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 3:46 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [hidden email]> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2BogpChJQXFCwavjPUSymecSHTqnhzwdxqD9yfUhn7Q%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C3b543bf01bc840e13f3008d8df258baf%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504699639178501%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=i2y8SAPoXZ7KUAL6iuPZ%2F2WhXz4oB2kRtRmXZRThmZU%3D&reserved=0 and include the link in your posting. ***** Dear all, Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. Mike |
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In reply to this post by mmodel
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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. ***** Talking about accurate focus, have you ever looked at how small particles go from dark to light? If they are scattered over the surface and you move the wedge apparatus sideways you can bring the band of light to dark transition move back to the center with the focus mechanism... Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 5:16 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [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. ***** Mark, Thank you for you patience with me. Well, yes, if I can reproducibly focus on something within much less than a micron - and if that something can be much less than a micron (so as to create a well-defined distance within << um) then... I will think about it. I am afraid the fringe experiment may not work because fringes are formed by reflection between two fixed pieces of glass, not between the glass and the objective. Best wishes Mike -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Cannell Sent: Thursday, March 4, 2021 11:55 AM To: [hidden email] Subject: Re: vertical stage travel ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe834df7c0dd4ffd51a108d8df2e5175%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504737478991307%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=h%2F5VDe51SYmJQ7aMCRcV%2F8yBOkKrEfoSRWCHfl6W6Qg%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe834df7c0dd4ffd51a108d8df2e5175%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504737479001305%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=c79xe0MpHsujLcSVuQH%2FsgYw0ZZ7gcLun6B6MCOqvgY%3D&reserved=0 and include the link in your posting. ***** Measure the thickness (csthick) of a coverslip and since that coverslip is placed to one side to create the wedge between the normal objective coverslip and a second test coverslip on top (width cswidth) you can use: dz=dx*sin(arctan(csheight/cswidth))) Where dz is the change in height to the test surface and dx is the lateral displacement. How you make the test coverslip surface visible is up to you -dirt, smoke particles, dried (fluorescent?) beads etc. If you want to get fancy you could even measure the change in height by observing the fringes in the wedge under monochromatic light ala Newton's fringe experiments __ HTH Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 4:41 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [hidden email]> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe834df7c0dd4ffd51a108d8df2e5175%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504737479001305%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=U%2B6Mg1lfRxm9nH84jU%2FfbIvLIJBBNvrBEbPRNw60lSc%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe834df7c0dd4ffd51a108d8df2e5175%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504737479001305%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=c79xe0MpHsujLcSVuQH%2FsgYw0ZZ7gcLun6B6MCOqvgY%3D&reserved=0 and include the link in your posting. ***** Thanks! But how would you determine the exact height of a coverslip within less then a micron on a widefield microscope? -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Mark Cannell Sent: Thursday, March 4, 2021 10:52 AM To: [hidden email] Subject: Re: vertical stage travel ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe834df7c0dd4ffd51a108d8df2e5175%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504737479001305%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=U%2B6Mg1lfRxm9nH84jU%2FfbIvLIJBBNvrBEbPRNw60lSc%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe834df7c0dd4ffd51a108d8df2e5175%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504737479001305%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=c79xe0MpHsujLcSVuQH%2FsgYw0ZZ7gcLun6B6MCOqvgY%3D&reserved=0 and include the link in your posting. ***** If a coverslip is placed on another at one side you've made an air wedge so that now lateral displacement can give small changes in Z position of the coverslip surface HTH Mark B. Cannell. Ph.D. FRSNZ FISHR Department of Physiology, Pharmacology & Neuroscience School of Medical Sciences University Walk Bristol BS8 1TD [hidden email] On 4/03/21, 3:46 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[hidden email] on behalf of [hidden email]> wrote: ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe834df7c0dd4ffd51a108d8df2e5175%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504737479001305%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=U%2B6Mg1lfRxm9nH84jU%2FfbIvLIJBBNvrBEbPRNw60lSc%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe834df7c0dd4ffd51a108d8df2e5175%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637504737479001305%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=c79xe0MpHsujLcSVuQH%2FsgYw0ZZ7gcLun6B6MCOqvgY%3D&reserved=0 and include the link in your posting. ***** Dear all, Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. Mike |
 
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Arnaud ROYON |
Re: vertical stage travel - VENDOR RESPONSE
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In reply to this post by mmodel
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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 Mike, You could use the 3D patterns in our calibration slides, for example the 3D crossing stairs. They consists of sub-micron empty cylinders embedded at different depths, like two crossing stairs. One of their applications is to control the axial distances. You can have a look at our applications guide (page 36) here: http://argolight.com/files/Argolight-solutions_Applications-guide.pdf Best regards, Arnaud Arnaud ROYON, Ph.D. CSO & CTO, Member of the Executive Board, Co-founder Argolight Cité de la Photonique, Bat. Elnath 11 avenue de Canteranne 33600 Pessac, FRANCE Email:[hidden email] Tel: (+33) 5 64 31 08 50 Web site:www.argolight.com Le 04/03/2021 à 16:43, MODEL, MICHAEL a écrit : > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > Post images onhttp://www.imgur.com and include the link in your posting. > ***** > > Dear all, > > Does anyone know of a way to verify the distance of a small vertical stage travel on a widefield scope? It is easy enough for large shifts by tens of microns, but we suspect that our small steps, such as by 1-5 microns, may be off. Of course, we can take 10 consecutive 3 um steps and check that they add up to 30, but maybe there is a better way? Would appreciate an advice. > > Mike |
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In reply to this post by mmodel
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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 you can borrow a Mirau-type interferometry microscope objective, then the wedge method is really easy, and it will work for reflection off surface. With a reasonably monochromatic light you will be able to see the fringes without any problem. I used the Nikon DI objective on our old Zeiss Axiophot some years ago and it worked well. You will also see very easily if your microscope needs a better vibration isolation table. Stan Vitha Microsocpy and Imaging Center Texas A&M University |
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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. ***** Thank you, Stan - That's about what I was looking for! In fact, such an objective may be available on campus. I only don't know if it will fit the thread Best wishes Mike -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Stanislav Vitha Sent: Friday, March 5, 2021 10:45 AM To: [hidden email] Subject: Re: vertical stage travel ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe782493af8249aa10db08d8dff795c0%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637505602334100576%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=mNU7YRNo%2F7iAa5vNQDDzDsnN9lkEB4PCow70y4GvRlw%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe782493af8249aa10db08d8dff795c0%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637505602334110572%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=ivQhEGzMTltujou9nLI19vdHwJz2LI%2FN2%2FQK38HStYo%3D&reserved=0 and include the link in your posting. ***** If you can borrow a Mirau-type interferometry microscope objective, then the wedge method is really easy, and it will work for reflection off surface. With a reasonably monochromatic light you will be able to see the fringes without any problem. I used the Nikon DI objective on our old Zeiss Axiophot some years ago and it worked well. You will also see very easily if your microscope needs a better vibration isolation table. Stan Vitha Microsocpy and Imaging Center Texas A&M University |
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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. ***** Would a test indicator not work in this case? One of these: https://www.amazon.com/Mitutoyo-Indicator-Horizontal-Metric-Diameter/dp/B003UATP1W?th=1 or similar has a position resolution of 1 µm. Accuracy is likely ~2x that or more, but you can likely resolve errors of a few microns, especially if you think there are quantization errors or similar at small step sizes. You could also use the linearity of a bounced return beam or the air wedge provided distance instead of the same from the test indicator. If there's an error at small step sizes, the response vs setting plot would deviate from a straight line at small step sizes. At this point you seem to want more to know if there is an error rather than its absolute magnitude. So a question wholly of precision, not accuracy, to determine that. The test indicator would be sufficient, assuming the stage will not deflect or fail to move normally with that extra force (stage can lift a well plate full of water, so it should be fine). Checking the linearity of the response of any method would minimize the impact of systematic errors in accuracy (such as the question on how to use the air wedge to derive absolute distance). These errors would show up as an offset or different slope in the response curve, neither of which is the first thing to measure when trying to establish a deviation from that linearity. Thanks, Rusty On Fri, Mar 5, 2021 at 9:16 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. > ***** > > Thank you, Stan - > That's about what I was looking for! In fact, such an objective may be > available on campus. I only don't know if it will fit the thread > > Best wishes > > Mike > > -----Original Message----- > From: Confocal Microscopy List <[hidden email]> On > Behalf Of Stanislav Vitha > Sent: Friday, March 5, 2021 10:45 AM > To: [hidden email] > Subject: Re: vertical stage travel > > ***** > To join, leave or search the confocal microscopy listserv, go to: > > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe782493af8249aa10db08d8dff795c0%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637505602334100576%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=mNU7YRNo%2F7iAa5vNQDDzDsnN9lkEB4PCow70y4GvRlw%3D&reserved=0 > Post images on > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7Cfe782493af8249aa10db08d8dff795c0%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637505602334110572%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=ivQhEGzMTltujou9nLI19vdHwJz2LI%2FN2%2FQK38HStYo%3D&reserved=0 > and include the link in your posting. > ***** > > If you can borrow a Mirau-type interferometry microscope objective, then > the wedge method is really easy, and it will work for reflection off > surface. With a reasonably monochromatic light you will be able to see the > fringes without any problem. I used the Nikon DI objective on our old > Zeiss Axiophot some years ago and it worked well. > You will also see very easily if your microscope needs a better vibration > isolation table. > > > > Stan Vitha > Microsocpy and Imaging Center > Texas A&M University > |
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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. ***** Thank you, Rusty. I decided to borrow a Mirau objective, as Stan suggested. If it does not fit our microscope, I will do it the easy way: large distances, such as 50 um, are easily measurable, and then I will just check if 5x10 = 50. Mike -----Original Message----- From: Confocal Microscopy List <[hidden email]> On Behalf Of Rusty Nicovich Sent: Saturday, March 6, 2021 1:13 PM To: [hidden email] Subject: Re: vertical stage travel ***** To join, leave or search the confocal microscopy listserv, go to: https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cmmodel%40KENT.EDU%7C8629d5ab3b0540c72d2108d8e1f53573%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637507790575714555%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=OizTYKbEVzZYuqAAvVGYTRjxScQEWERvlJjhXoxR0FM%3D&reserved=0 Post images on https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C8629d5ab3b0540c72d2108d8e1f53573%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637507790575714555%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=38hTVBVKltMDbNKGW0UYBzuYPjVe1QfzV1y5Ua36P5A%3D&reserved=0 and include the link in your posting. ***** Would a test indicator not work in this case? One of these: https://nam11.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.amazon.com%2FMitutoyo-Indicator-Horizontal-Metric-Diameter%2Fdp%2FB003UATP1W%3Fth%3D1&data=04%7C01%7Cmmodel%40KENT.EDU%7C8629d5ab3b0540c72d2108d8e1f53573%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637507790575714555%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=kX1%2BdmqdxeEy2uKSDRuW8W1VTelb5Hd94RYv7mBdVms%3D&reserved=0 or similar has a position resolution of 1 µm. Accuracy is likely ~2x that or more, but you can likely resolve errors of a few microns, especially if you think there are quantization errors or similar at small step sizes. You could also use the linearity of a bounced return beam or the air wedge provided distance instead of the same from the test indicator. If there's an error at small step sizes, the response vs setting plot would deviate from a straight line at small step sizes. At this point you seem to want more to know if there is an error rather than its absolute magnitude. So a question wholly of precision, not accuracy, to determine that. The test indicator would be sufficient, assuming the stage will not deflect or fail to move normally with that extra force (stage can lift a well plate full of water, so it should be fine). Checking the linearity of the response of any method would minimize the impact of systematic errors in accuracy (such as the question on how to use the air wedge to derive absolute distance). These errors would show up as an offset or different slope in the response curve, neither of which is the first thing to measure when trying to establish a deviation from that linearity. Thanks, Rusty On Fri, Mar 5, 2021 at 9:16 AM MODEL, MICHAEL <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists > .umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cm > model%40KENT.EDU%7C8629d5ab3b0540c72d2108d8e1f53573%7Ce5a06f4a1ec44d01 > 8f73e7dd15f26134%7C1%7C0%7C637507790575714555%7CUnknown%7CTWFpbGZsb3d8 > eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1 > 000&sdata=OizTYKbEVzZYuqAAvVGYTRjxScQEWERvlJjhXoxR0FM%3D&reser > ved=0 Post images on > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C8629d5ab3b0540c72d2108d8e1f53573%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C637507790575724548%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=3gxyLkHRwjvrDJD4sY7El%2FTYtU6D1UsXWwzi%2F6AvQjM%3D&reserved=0 and include the link in your posting. > ***** > > Thank you, Stan - > That's about what I was looking for! In fact, such an objective may be > available on campus. I only don't know if it will fit the thread > > Best wishes > > Mike > > -----Original Message----- > From: Confocal Microscopy List <[hidden email]> On > Behalf Of Stanislav Vitha > Sent: Friday, March 5, 2021 10:45 AM > To: [hidden email] > Subject: Re: vertical stage travel > > ***** > To join, leave or search the confocal microscopy listserv, go to: > > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists > .umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Cm > model%40KENT.EDU%7C8629d5ab3b0540c72d2108d8e1f53573%7Ce5a06f4a1ec44d01 > 8f73e7dd15f26134%7C1%7C0%7C637507790575724548%7CUnknown%7CTWFpbGZsb3d8 > eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1 > 000&sdata=LWgMqPmFceTKE8Nbdyhd98xPRi15gnCehBr4%2Fwutevo%3D&res > erved=0 > Post images on > https://nam11.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.i > mgur.com%2F&data=04%7C01%7Cmmodel%40KENT.EDU%7C8629d5ab3b0540c72d2 > 108d8e1f53573%7Ce5a06f4a1ec44d018f73e7dd15f26134%7C1%7C0%7C63750779057 > 5724548%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLC > JBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=3gxyLkHRwjvrDJD4sY7El%2F > TYtU6D1UsXWwzi%2F6AvQjM%3D&reserved=0 > and include the link in your posting. > ***** > > If you can borrow a Mirau-type interferometry microscope objective, > then the wedge method is really easy, and it will work for reflection > off surface. With a reasonably monochromatic light you will be able to > see the fringes without any problem. I used the Nikon DI objective on > our old Zeiss Axiophot some years ago and it worked well. > You will also see very easily if your microscope needs a better > vibration isolation table. > > > > Stan Vitha > Microsocpy and Imaging Center > Texas A&M University > |
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*****
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. ***** I forgot to state the obvious that for the Mirau objective, you will need a reflected light illumination setup. Stan Texas A&M University Microscopy and Imaging Center |
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