Posted by
Moulding, Dale on
Apr 26, 2021; 5:16pm
URL: http://confocal-microscopy-list.275.s1.nabble.com/widefield-autofluorescence-in-unlabeled-cells-a-filter-mystery-tp7592150p7592153.html
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Hi Doug,
Does it have an LED for transmitted light? Is it the LED shining back at the sample? I had this problem on an old scope, when I put my hand between condenser and the 'fluorescence' went away. Fixed with a green interference filter in the transmitted light path.
Cheers
Dale
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Sent: 26 April 2021 17:48
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Subject: widefield autofluorescence in unlabeled cells - a filter mystery?
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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
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