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Some people in our lab use Alexa 546 because they want to use the same antibody they use for FACS, but in general we recommend Alexa 568 which we use successfully with diode laser excitation at 532 (TIRF and dSTORM/PALM/GSD), 543 (confocal) and 561 nm (widefield and TIRF).
________________________________________________________
Michael Cammer, Assistant Research Scientist
Microscopy Core, NYU Langone Medical Center & Skirball Institute of Biomolecular Medicine
Cell: (914) 309-3270 Microscopy Lab: (212) 263-7099 Dustin Lab: (212) 263-3208
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Xuejun SUN
Sent: Thursday, July 04, 2013 11:17 AM
To: [log in to unmask]
Subject: Re: Alexa Fluor 532 with 543nm laser
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Hi, Scott,
I have not tried Alexa 532. However, we did have a lot of experiences with Alexa 546. Put a long story short, I was told (by then a Mol. Probe person) that it is a "bad" dye to work with as it is almost incompatible with any antifading agent and it works best without any antifading product. So we switched to Alexa 555 or 568 and are much happier. Maybe you could try a mounting media without antifade?
Good luck,
Xuejun
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Scott Wong
Sent: Tuesday, July 02, 2013 5:08 PM
To: [log in to unmask]
Subject: Alexa Fluor 532 with 543nm laser
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Hi everyone,
I was wondering if anyone has successfully used the Alexa Fluor 532 secondary to view tyrosine hydroxylase in dopaminergic neurons. I'll be imaging it using a 543nm laser with a Nikon confocal microscope. Based on its excitation wavelength it seems to be an ideal match but I don't any experience with it.
If not, can anyone comment on its anti-quenching properties? I'm currently using the Alexa Fluor 546 for the same purpose but it quenches almost unusably fast with every mounting media I've tried.
Thanks for the help,
Scott Wong
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