Autofluorescence in silk

[Paul Rigby-2]

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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

 

 

[Guy Cox]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

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
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
______________________________________________
Phone [phone]     Fax [phone]
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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

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

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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[Sebastian Frische]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi

Silk is an aniosotropic protein fibre, so in polarized light it gives strong birefringence. I wonder if the use of a laser may be enough to elicit this to level where the material "light up".

best wishes

Sebastian

Den 15/08/2008 kl. 11.04 skrev Paul Rigby:

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

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

 

 

Lektor, PhD

Sebastian Frische

Anatomisk Institut

Aarhus Universitet

Universitetsparken Bygn. 1234

8000 Århus C

Danmark

tlf [phone]

fax + 45 86198664

[hidden email]

[Zucker.Robert]

Hi Paul
How are you ?
playing with Zeiss 510  unmixing and it is great.
what are you buying? I have some views on the subject. -
did you have luck with the detergent?
I am trying it again.
bob


Robert M. Zucker, PhD
U.S. Environmental Protection Agency
Office of Research and Development
National Health and Environmental Effects Research Laboratory.
Reproductive Toxicology Division
Telephone: [phone]   Fax: [phone]
e-mail: [hidden email]

Mail address: USEPA,ORD,NHEERL,RTD
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
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