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Just to correct my mistake : the laser lines I tested are 532 nm (Leica
SPE) and 543 nm (Zeiss LSM 780), not 561 nm.

Christophe
Le 5 juil. 2013 23:11, "Christophe Leterrier" <
[log in to unmask]> a écrit :

> 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
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
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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
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
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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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>