Strange Photo conversion

Posted by Cameron Nowell-3 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Strange-Photo-conversion-tp7586797.html

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi List,

We are trying to get some experiments up and running and have hit a bizarre
artifact with Cy5 getting photo converted and becoming very very bright.

The experimental desing is as follows
- fluorodish or ibidi channel slide
- coated with Poly-D-Lysine in PBS
- Washed PBS
- Washed Bi-carbonate pH 8.6
- Couple Cy 5 succinamide ester bi-carb pH 8.6
- Wash Bi-carb 8.6
- Wash PBS

IMaging on either a TIRF or confocal system we get a rapid and robust
increase in fluorescene in the Cy5 channel. Exited at any wavelegth (405,
488 or 647).

Any ideas on what could be going on?

Cheers

Cam


--

*Cameron J. Nowell*

Head



Imaging, FACS and Analysis Core

Monash Institute of Pharmaceutical Sciences

Monash University

399 Royal Parade

Parkville, VIC, 3052

Australia



*Email:* [hidden email]

*Phone: *[phone]



*LinkedIn: *Profile <http://au.linkedin.com/pub/cameron-nowell/23/57/884/>

*Research Gate: * Profile
<http://www.researchgate.net/profile/Cameron_Nowell>

*Google Scholar:* Profile
<https://scholar.google.com/citations?user=qqQfgZIAAAAJ&hl=en>

*PubMed Bibliography: *Profile
<http://www.ncbi.nlm.nih.gov/sites/myncbi/1TK3WeparcvQr/bibliography/47922177/public/?sort=date&direction=ascending>