Re: Fluorophore excitable with 405nm and 488nm lines

Posted by Craig Brideau on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Fluorophore-excitable-with-405nm-and-488nm-lines-tp6432486p6445988.html

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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 <[hidden email]> wrote:

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> 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 [phone]     Fax [phone]
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>      http://www.guycox.net
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>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Cammer, Michael
> Sent: Friday, 3 June 2011 9:00 PM
> To: [hidden email]
> Subject: Re: Fluorophore excitable with 405nm and 488nm lines
>
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> To join, leave or search the confocal microscopy listserv, go to:
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> 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: [phone]  Cell: [phone]
>
> ________________________________________
> From: Confocal Microscopy List [[hidden email]] On
> Behalf Of Guy Cox [[hidden email]]
> Sent: Thursday, June 02, 2011 11:32 PM
> To: [hidden email]
> 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 [phone]     Fax [phone]
>             Mobile [phone]
>
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