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I would like to thank everyone for their suggestions. I will try some out next week, and I'll let you know what worked for me.
Daniel
----- Original Message -----
From: Craig Brideau <[log in to unmask]>
Date: Monday, June 6, 2011 20:10
Subject: Re: Fluorophore excitable with 405nm and 488nm lines
To: [log in to unmask]
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> That's pretty neat Guy! Where on earth would you find
> synthetic emerald
> though?
>
> Craig
>
>
>
> On Sat, Jun 4, 2011 at 2:05 AM, Guy Cox
> <[log in to unmask]> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Hang on, the requirement was for a sample which would measure the
> > RELATIVE intensities of 505 and 488nm lasers. Mild
> bleaching shouldn't
> > affect that. If a bit is seriously bleached, use another
> spot. A quick
> > scan at very low power will reveal that. You're doing
> pretty well if
> > you can bleach an entire slide in a lifetime - and it you do, the
> > replacement is free. I've never succeeded in bleaching
> any part but I
> > have managed to laser-ablate spots on the surface! As
> for RI mismatch -
> > well of course there is. I'll bet there is with uranyl
> glass too (but
> > in the opposite direction). But the killer with the U
> glass is that you
> > can't actually see it very well in a scanned image. If
> you really want
> > a solid-state unbleachable sample get a slice of
> emerald. I've never
> > actually tried it at 405nm but it's so bright at 488 I'm sure
> it would
> > work. Unless you have a very generous lab budget I would
> recommend the
> > synthetic version over the natural one!
> >
> > Guy
> >
> >
> > Optical Imaging Techniques in Cell Biology
> > by Guy Cox CRC Press / Taylor & Francis
> > http://www.guycox.com/optical.htm
> > ______________________________________________
> > Associate Professor Guy Cox, MA, DPhil(Oxon)
> > Australian Centre for Microscopy & Microanalysis,
> > Madsen Building F09, University of Sydney, NSW 2006
> >
> > Phone +61 2 9351 3176 Fax +61 2 9351 7682
> > Mobile 0413 281 861
> > ______________________________________________
> > http://www.guycox.net
> >
> >
> > -----Original Message-----
> > From: Confocal Microscopy List
> [mailto:[log in to unmask]]> On Behalf Of Cammer,
> Michael> Sent: Friday, 3 June 2011 9:00 PM
> > To: [log in to unmask]
> > Subject: Re: Fluorophore excitable with 405nm and 488nm lines
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Depending on your tolerance, they may or may not be
> stable. I use these
> > all the time for aligning the microscope because they are
> great for
> > this, although there may be an issue with refractive index mismatch
> > especially with the newer TIRF objectives, but I'm not so sure
> as a
> > reproducible sample.
> > Please see
> > http://www.einstein.yu.edu/aif/instructions/fluor-ref-
> slides/01.htm and
> >
> http://www.einstein.yu.edu/aif/instructions/zeiss/liveduo/Zaxis_bleachin> gtest/index.htm
> > for two examples of bleaching.
> > -Michael
> >
> > _________________________________________
> > Michael Cammer, Assistant Research Scientist
> > Skirball Institute of Biomolecular Medicine
> > Lab: (212) 263-3208 Cell: (914) 309-3270
> >
> > ________________________________________
> > From: Confocal Microscopy List
> [[log in to unmask]] On
> > Behalf Of Guy Cox [[log in to unmask]]
> > Sent: Thursday, June 02, 2011 11:32 PM
> > To: [log in to unmask]
> > Subject: Re: Fluorophore excitable with 405nm and 488nm lines
> >
> > I just checked out a Chroma green fluorescent plastic slide -
> it seemed
> > to fluoresce pretty well with both uv and blue LEDs so surely that
> > should work well enough. Doesn't have the ultra-long
> lifetime problem
> > of uranyl glass. Reproducible, robust and best of all free!
> >
> > Guy
> >
> > Optical Imaging Techniques in Cell Biology
> > by Guy Cox CRC Press / Taylor & Francis
> > http://www.guycox.com/optical.htm
> > ______________________________________________
> > Associate Professor Guy Cox, MA, DPhil(Oxon)
> > Australian Centre for Microscopy & Microanalysis,
> > Madsen Building F09, University of Sydney, NSW 2006
> >
> > Phone +61 2 9351 3176 Fax +61 2 9351 7682
> > Mobile 0413 281 861
> >
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Daniel Gitler, Ph.D.
Department of Physiology and Neurobiology
Faculty of Health Sciences
Ben Gurion University of the Negev
Beer-Sheva 84105
Israel
Tel: +972-8-6477345
Cell: +972-54-2110100
Fax: + 972-8-6477628
http://web2.bgu.ac.il/physiology/faculty-members/daniel-gitler/
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