Posted by
Carol Heckman on
URL: http://confocal-microscopy-list.275.s1.nabble.com/widefield-autofluorescence-in-unlabeled-cells-a-filter-mystery-tp7592150p7592157.html
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopyPost images on
http://www.imgur.com and include the link in your posting.
*****
Unlabeled cells do fluoresce in the yellow-green range. I have always thought it is because they have soluble molecules such as flavenoids in the cytoplasm. If the cells are permeabilized, for example, to use them in immunofluorescence procedures, there is much less of this background.
Carol Heckman
Bowling Green State University
________________________________
From: Confocal Microscopy List <
[hidden email]> on behalf of Cromey, Douglas W - (dcromey) <
[hidden email]>
Sent: Monday, April 26, 2021 12:47 PM
To:
[hidden email] <
[hidden email]>
Subject: [EXTERNAL] widefield autofluorescence in unlabeled cells - a filter mystery?
*****
To join, leave or search the confocal microscopy listserv, go to:
https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Flists.umn.edu%2Fcgi-bin%2Fwa%3FA0%3Dconfocalmicroscopy&data=04%7C01%7Checkman%40BGSU.EDU%7Cf170296dad8f4744d55f08d908d3772c%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C0%7C637550526594680955%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=dCTyUY%2F8ngiEn%2FVHtKCIEu74ht85UrmxHLeIHH%2FWxIA%3D&reserved=0Post images on
https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.imgur.com%2F&data=04%7C01%7Checkman%40BGSU.EDU%7Cf170296dad8f4744d55f08d908d3772c%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C0%7C637550526594680955%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=2losqYJ9Z9qpo5yOu5KBsmNg4AM6XWpo5p1oBLSDCTg%3D&reserved=0 and include the link in your posting.
*****
A colleague asked me to take a look at their widefield microscope. It is an inverted microscope with a 100W Hg source, excitation filter wheel, a couple of choices for dichroics in the microscope filter changer and a filter wheel in front of the camera. They are seeing unlabeled cells fluoresce green (FITC/GFP set) with an otherwise black background where there are no cells. The microscope is approximately 15 years old.
My guess is that the excitation filters have failed (or are failing) after being on the receiving end of a 100W Hg lamp for all this time. Any other thoughts?
Doug
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Associate Scientific Investigator
Dept. of Cellular & Molecular Medicine, University of Arizona
Life Sciences North, Room 463
1333 N. Martin Ave, Tucson, AZ 85721 USA
office: LSN 463 email:
[hidden email]<mailto:
[hidden email]>
voice: 520-626-2824 fax: 520-626-2097
UA Microscopy Alliance -
https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fmicroscopy.arizona.edu%2F&data=04%7C01%7Checkman%40BGSU.EDU%7Cf170296dad8f4744d55f08d908d3772c%7Ccdcb729d51064d7cb75ba30c455d5b0a%7C1%7C0%7C637550526594680955%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&sdata=qV%2BaW43NPHbyhJnzTKGCzr2KRicX1uPf%2BxkC2%2Fnx3yg%3D&reserved=0A collaborative effort to bring information about shared microscopy
facilities to the University of Arizona and the community.