Confocal Microscopy List

Re: Autofluorescence in silk

Posted by Guy Cox on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Autofluorescence-in-silk-tp725908p725938.html

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If these are fixed cells I'd assume it is aldehyde based autofluorescence

since silk is a protein. If they are living cells I've no idea. But you could try

to attack the silk with a protease, which at modest concentrations

probably wouldn't get into living cells.

Guy

Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
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Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
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From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Paul Rigby
Sent: Friday, 15 August 2008 7:05 PM
To: [hidden email]
Subject: Autofluorescence in silk

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Hi All,

One of our users has a question about silk and autofluorescence –

“I am culturing keratinocyte cells on several different varieties of silk (from silk worms) and have discovered that in general, the silk exhibits quite strong autofluorescence (particularly at 488nm excitation). Does anybody have any suggestions to help quench this autofluorescence?”

The normal treatments (Sudan black, Pontamine Sky Blue, Sodium Borohydride, Ammonium Chloride), for one reason or another, are not suitable or relevant in our system. I have suggested trying spectral unmixing as a possible solution, but if there are any other suggestions…..

Does anyone know what the chemistry is in silk that might lead to the autofluorescence?

All comments/thoughts appreciated.

Regards

Paul

Dr Paul Rigby

Senior Lecturer

Centre for Microscopy, Characterisation and Analysis (M510)

The University of Western Australia

35 Stirling Highway

Crawley WA 6009

Ph (61-8) 9346 2819

Fx (61-8) 9346 3469

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