poly-L-lysine : internal reflection

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Chris Katnik Chris Katnik
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poly-L-lysine : internal reflection

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Is there ever a problem with using ploy-L-lysine coated coverslips when doing
confocal imaging of FITC or Alexa Fluor 555 stained cells and internal reflection
of the laser light?
CONFOCAL TL Andresen CONFOCAL TL Andresen
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Re: poly-L-lysine : internal reflection

Search the CONFOCAL archive at
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Hi Chris

Yes, when you stain with FITC it will also react with poly-lysine. And
you will naturally get huge background fluorescence. You need to use a
fluorofor that does not contain a isothiocyante group (which mean TRITC
is no good either).

Best regards, Thomas

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Chris Katnik
Sent: Wednesday, August 27, 2008 5:12 PM
To: [hidden email]
Subject: poly-L-lysine : internal reflection

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

Is there ever a problem with using ploy-L-lysine coated coverslips when
doing
confocal imaging of FITC or Alexa Fluor 555 stained cells and internal
reflection
of the laser light?
Ignatius, Mike Ignatius, Mike
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Re: poly-L-lysine : internal reflection

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Hi Chris,

We were tossing this issue around here and thought one recommendation
was warranted.

We make a product called Image-iT FX Signal Enhancer that works nicely
for blocking non-specific interactions of dyes and dye-labeled
bioconjugates to things with net positive charges like poly-lysine.  

There is a complete listing of dyes that have been tested at
http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applicat
ions/Cell-and-Tissue-Analysis/Cellular-Imaging/Fluorescence-Microscopy-a
nd-Immunofluorescence-IF/Image-iT-FX-Kits.reg.us.html

Commercial Bias acknowledged....

Mike Ignatius

Molecular Probes/Invitrogen

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of CONFOCAL TL Andresen
Sent: Wednesday, August 27, 2008 8:43 AM
To: [hidden email]
Subject: Re: poly-L-lysine : internal reflection

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

Hi Chris

Yes, when you stain with FITC it will also react with poly-lysine. And
you will naturally get huge background fluorescence. You need to use a
fluorofor that does not contain a isothiocyante group (which mean TRITC
is no good either).

Best regards, Thomas

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Chris Katnik
Sent: Wednesday, August 27, 2008 5:12 PM
To: [hidden email]
Subject: poly-L-lysine : internal reflection

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

Is there ever a problem with using ploy-L-lysine coated coverslips when
doing
confocal imaging of FITC or Alexa Fluor 555 stained cells and internal
reflection
of the laser light?