***** 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 <[log in to unmask]> On Behalf Of Shuce Zhang Sent: Thursday, March 4, 2021 11:00 AM To: [log in to unmask] 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 [log in to unmask] | 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 <[log in to unmask]> 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 > > [log in to unmask] > > > > On 4/03/21, 3:46 PM, "Confocal Microscopy List on behalf of MODEL, MICHAEL" <[log in to unmask] on behalf of [log in to unmask]> 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 >
