http://confocal-microscopy-list.275.s1.nabble.com/Syto-dyes-for-plant-nuclei-in-living-cells-tp592149p592158.html
I only used the red Sytos once on live brain slices. The red Syto
membrane compartments on their way to the nucleus. You have to wait
> Search the CONFOCAL archive at
>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>
> Hi John and Tobias,
>
> This may be a completely unhelpful response because I was not
> working on plants and I was looking for dyes in the red/far red
> range, not blue. However, a few years ago I tested a whole range of
> the red Syto dyes to try to use them to follow cell division using
> live cell imaging (together with a GFP construct) and I found
> something very interesting with Syto 61. Although I had the same
> problem as Tobias when I imaged through Texas Red filter sets -
> i.e. the mitochondria obscured the nuclear signal - I found that if
> I acquired through the far red (Cy5) filters the signal was now
> predominantly nuclear. The longer I left it, the more the nuclear
> signal seemed to intensify while the remaining mitochondrial signal
> soon bleached away. I called Mol. Probes to ask them about this and
> they said that the dye becomes red-shifted when it's highly
> concentrated - which fits with the fact that the mitotic cells were
> brightest. Anyway, unlike most of the nuclear dyes it didn't
> affect cell division so I could image away for hours/days.
>
> One slight problem - the signal and localisation appeared to be
> highly cell specific. It was great in MDCK cells but I seem to
> remember I couldn't get the same effect in HeLas. Still - might be
> worth a shot if a far red dye would do?
>
> Best wishes,
> Alison
>
>
> Tobias Baskin wrote:
>> Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/
>> cgi-bin/wa?S1=confocal
>> John,
>> One or two years ago, we got a group of syto dyes for this
>> very purpose, imaging in arabidopsis roots. We were hoping to
>> image DNA along with a GFP reporter and therefore we tested sytos
>> that have long wavelength emission (we were working on confocal,
>> without ability to do uv excitation). None of them worked for us.
>> They tended to be either toxic or dim. The other thing we noticed
>> was that because the cells are crammed with mitochondria that move
>> all over the place, it was difficult to follow nuclear DNA against
>> this hive of mitochondrial motion. But we never tested blue ones.
>> Good luck,
>> Tobias
>>> Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/
>>> cgi-bin/wa?S1=confocal
>>>
>>> Do any of the plant biologists out there have experience using
>>> the Syto dyes from Molecular Probes to stain nuclei of living
>>> cells. In particular, they list 5 different dyes in the blue
>>> emission range (Syto 40-45). Syto 42 looks good to me on paper.
>>> What do you think. Thanks, John.
>>> --
>>> Runions signature
>>> *********************************
>>> C. John Runions, Ph.D.
>>> School of Life Sciences
>>> Oxford Brookes University
>>> Oxford, UK
>>> OX3 0BP
>>>
>>> email:
[hidden email] <mailto:
[hidden email]>
>>> phone: +44 (0) 1865 483 964
>>> Runions' lab web site <
http://www.brookes.ac.uk/lifesci/runions/
>>> HTMLpages/index.html%21>
>>> Visit The Illuminated Plant Cell <
http://www.illuminatedcell.com/
>>> ER.html> dot com
>>> Oxford Brookes Master's in Bioimaging with Molecular Technology
>>> <
http://www.brookes.ac.uk/studying/courses/postgraduate/2007/bmt>
>> --
>> _ ____ __ ____ / \ / /
>> \ / \ \ Tobias I. Baskin
>> / / / / \ \ \ Biology Department
>> /_ / __ /__ \ \ \__ 611 N. Pleasant St.
>> / / / \ \ \ University of
>> Massachusetts
>> / / / \ \ \ Amherst, MA,
>> 01003
>> / / ___ / \ \__/ \ ____
>>
http://www.bio.umass.edu/biology/baskin/>> Voice: 413 - 545 - 1533 Fax:/ 413 - 545 - 3243/
>
> --
> Alison J. North, Ph.D.,
> Research Assistant Professor and
> Director of the Bio-Imaging Resource Center,
> The Rockefeller University,
> 1230 York Avenue,
> New York,
> NY 10065.
> Tel: office ++ 212 327 7488
> Tel: lab ++ 212 327 7486
> Fax: ++ 212 327 7489