Chris Katnik |
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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? |
CONFOCAL TL Andresen |
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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? |
Ignatius, Mike |
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal 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? |
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