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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hello Listers, I have seen a lot of posts discussing variouos nuclear stains, and I have a question for you all. I work with a Nikon Confocal system that has 3 lasers, a 405, 488 and 543. I am having trouble finding a good nuclear stain that I can use on living cells that will work with these lasers. I have been using hoescht, but it is not optimal excited with the 405 laser. Are there any alternatives that anyone knows of that can be used on living cells??? Thanks for your help! Sarah Aubut |
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The latest issue of Cytometry has several wonderful reviews of DNA dyes
(Syto and DRAQ5) and their properties.....and amazingly it is
available ON-LINE at
http://www3.interscience.wiley.com/journal/33945/home At 08:57 AM 6/11/2008, you wrote: Search the CONFOCAL archive at Cytometry Facility Huck Institute of the Life Sciences 319 Life Sciences Building Penn State University University Park, PA 16802 http://www.huck.psu.edu/facilities/cytometry-up/ 814-863-2762 |
In reply to this post by Sarah Aubut
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Sarah, On that instrument, you likely could utilize Draq5. For multi-color imaging, you could optimally utilize your 543 laser for Draq5 excitation and hopefully in your case a long-pass emission filter is available. With those lasers you likely don't have a band-pass filter that is perfect for Draq5, but different versions of long-pass filters associated with your 543 line would work fine for this scenario. If you have any questions about this, feel free to contact me off-list and I'd be happy to help further. Cheers, -- Samuel A. Connell Director of Light Microscopy Cell & Tissue Imaging Center St. Jude Children's Research Hospital 332 North Lauderdale St., E7061 Memphis, TN 38105-2794 (901) 495-2536 [hidden email] On 6/11/08 7:57 AM, "Sarah Aubut" <[hidden email]> wrote: Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hello Listers, I have seen a lot of posts discussing variouos nuclear stains, and I have a question for you all. I work with a Nikon Confocal system that has 3 lasers, a 405, 488 and 543. I am having trouble finding a good nuclear stain that I can use on living cells that will work with these lasers. I have been using hoescht, but it is not optimal excited with the 405 laser. Are there any alternatives that anyone knows of that can be used on living cells??? Thanks for your help! Sarah Aubut |
In reply to this post by Sarah Aubut
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal you can use ethidium bromide with your laser Sarah Aubut wrote: >Search the CONFOCAL archive at >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > >Hello Listers, > >I have seen a lot of posts discussing variouos nuclear stains, and I have a >question for you all. >I work with a Nikon Confocal system that has 3 lasers, a 405, 488 and 543. I >am having trouble finding a good nuclear stain that I can use on living >cells that will work with these lasers. I have been using hoescht, but it is >not optimal excited with the 405 laser. Are there any alternatives that >anyone knows of that can be used on living cells??? > >Thanks for your help! >Sarah Aubut > > > > |
Kilgore, Jason |
In reply to this post by Elaine Kunze
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**Industry Response**
Another option to the dye options that have been
recommended is Hoechst 34580, which is readily excited at 405nm and can be used
for both live and fixed-cell nuclear labeling the same as the UV-excited Hoechst
dyes.
The excitation is between ~360-430nm. Emission is
between ~460-610nm.
This is sold by Molecular Probes / Invitrogen, product
H21486:
The best live-cell green-fluorescent we sell is SYTO 14,
product S7576:
Cheers,
Jason
Jason A. Kilgore
Invitrogen Corporation
Molecular Probes Labeling and Detection
Technologies
Technical Service Scientist
Eugene, OR 97402-9132
phone: (541) 335-0353
fax: (541) 335-0238
Please visit our website: www.probes.invitrogen.com From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Elaine Kunze Sent: Wednesday, June 11, 2008 6:06 AM To: [hidden email] Subject: Re: Nuclear Stains http://www3.interscience.wiley.com/journal/33945/home At 08:57 AM 6/11/2008, you wrote: Search the CONFOCAL archive at Cytometry Facility Huck Institute of the Life Sciences 319 Life Sciences Building Penn State University University Park, PA 16802 http://www.huck.psu.edu/facilities/cytometry-up/ 814-863-2762 |
In reply to this post by Vickery Trinkaus-Randall
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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi, If not already mentionned, here is an article comparing various nuclear dyes. I've not had super results with Draq5 on live cells. Lots of non-specific staining. M http://www3.interscience.wiley.com/journal/112783064/abstract -----Message d'origine----- De : Vickery Trinkaus-Randall [mailto:[hidden email]] Envoyé : 11 juin 2008 09:54 À : [hidden email] Objet : Re: Nuclear Stains Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal you can use ethidium bromide with your laser Sarah Aubut wrote: >Search the CONFOCAL archive at >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > >Hello Listers, > >I have seen a lot of posts discussing variouos nuclear stains, and I >have a question for you all. >I work with a Nikon Confocal system that has 3 lasers, a 405, 488 and >543. I am having trouble finding a good nuclear stain that I can use on >living cells that will work with these lasers. I have been using >hoescht, but it is not optimal excited with the 405 laser. Are there >any alternatives that anyone knows of that can be used on living cells??? > >Thanks for your help! >Sarah Aubut > > > > |
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