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From my parallel tests (but I don't have quantified data), Alexa 555
coupled secondary antibodies give a brighter signal than 546 and 568 ones
with both 561 and 532 nm laser lines on our confocals.

Christophe
Le 5 juil. 2013 18:04, "Cammer, Michael" <[log in to unmask]> a
écrit :

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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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