Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Glen,
but what do you do if you have 4 or 5 stainings in your sample - you
have to involve blue. Or you select a green look-up-table for Alexa 405.
Or for quantitative approaches, one can use spectrum or fire lut.
Fiddling with the histogram is not always a good idea, at least when it
has to be scientific.
Michael
Glen MacDonald wrote:
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> All of the Alexa fluor conjugates have a tendency to stick
> non-specifically via charge interactions of the anionic dye moiety. the
> Image-iT fx does helps greatly, especially in thick samples.
>
> In regard to using the blue fluorophores for the more prominent targets
> in the tissue, the human visual system is only weakly responsive to blue
> wavelengths and lacks blue-sensitive cones in the fovea, the region of
> greatest visual acuity. In general, if the smallest or rarest bits get
> the green, red or gray channel, they are noticeable with less histogram
> manipulation than if the smallest bits are viewed in the blue channel.
>
> Regards,
> Glen
>
>
>
> Glen MacDonald
> Core for Communication Research
> Virginia Merrill Bloedel Hearing Research Center
> Box 357923
> University of Washington
> Seattle, WA 98195-7923 USA
> (206) 616-4156
> [log in to unmask]
>
> ******************************************************************************
>
> The box said "Requires WindowsXP or better", so I bought a Macintosh.
> ******************************************************************************
>
>
>
> On Mar 6, 2008, at 7:09 AM, Junya HIROI wrote:
>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Christophe,
>>
>> I was also looking for a fluorescent dye for 405 nm excitation, and I
>> compared Alexa 405
>> with Pacific Blue. Both dyes bleached to some extent with
>> ProlongGold, but Pacific Blue was
>> absolutely brighter than Alexa 405, when compared at the same
>> dilution. So, I recommend
>> Pacific Blue, although Molecular Probes sells Pacific Blue for flow
>> cytometry.
>>
>> I found another problem on Alexa 405: it nonspecifically binds to
>> nuclei ! I asked Molecular
>> Probes/Invitrogen about this problem, and they recommended me to use
>> their blocking
>> solution, Image-iT FX Signal Enhancer. It actually worked well, but
>> Pacific Blue does not have
>> such a problem, so that is another reason I like Pacific Blue.
>>
>> Junya
>>
>> --
>> _____________________________________
>> 廣井準也
>> 聖マリアンナ医科大学 解剖学教室
>> 〒216-8511 川崎市宮前区菅生2-16-1
>> tel: 044-977-8111 内線3629
>> fax: 044-976-7083
>> e-mail: [log in to unmask]
>> _____________________________________
|
