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Hello all;
I was also impressed with the STED microscope at SFN. Great
improvement in resolution. However, as Martin said, it only works with
two dyes now (although someone brought a sample with them to the demo,
and found that their dye also worked...now there are three), and only
one dye at a time. The system is only good for fixed samples...it didn't
appear to even be useful for cell surface labeling of live cells.
Because of the requirement of having two lasers at different wavelengths
perfectly aligned, penetration depth was limited to 15-20 microns. I'm
still sending them some samples from our labs, but I too am waiting for
the "second-generation" STED before I send my PO.
Kathy Spencer, PhD
Microscopy Core Manager
ICND
The Scripps Research Institute
La Jolla, CA 92037
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Armstrong, Brian
Sent: Thursday, November 08, 2007 12:38 PM
To: [log in to unmask]
Subject: Re: Leica STED machine
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Thank you for sharing your experience.
I talked with Leica at SFN, but did not arrange a specific demo time
because I was previously aware of the $1.3 mil price tag. I attended a
"Super-Resolution Microscopy" meeting held by Leica at UCLA where the
experts from Germany on STED and 4Pi spoke. However, I was not aware
that the instrument was so particular about the fluorophores.
My take is that Zeiss (Big-Blue) does not like to be outdone and
certainly the alliance of Stefan Hell with Leica could cause them
concern. I have also heard a rumor that Zeiss is coming out with a
STED-like instrument, or at least a sub-diffration limit microscopy
system.
Moreover, it seems that Leica or Zeiss could make a system that utilizes
sub-diffraction techniques such as FPALM or STORM that could be
performed via software after imaging (similar to having a dedicated
deconvolution microscopy system).
As for my prediction for the "cutting edge time" for the STED
microscope???
I have no idea.
Brian D Armstrong PhD
Light Microscopy Core Manager
Beckman Research Institute
City of Hope
1450 E Duarte Rd
Duarte, CA 91010
626-359-8111 x62872
http://www.cityofhope.org/SharedResources/LightMicroscopy
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Martin Wessendorf
Sent: Thursday, November 08, 2007 11:23 AM
To: [log in to unmask]
Subject: Leica STED machine
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Hey folks!
I just got back from the Society for Neuroscience meeting in San Diego,
where Leica was giving a demo of it's Stimulated Emission Depletion
(STED) instrument.
Seeing it was an interesting experience. The microscope appeared to
improve resolution of some specimens (specifically, histones) by a
factor of probably 5-fold--there was a very pronounced improvement and
unless their scale bar was lying, they seemed to be down into the 50 nm
range. It did not seem to offer as much improvement on their muscle
specimen, and photobleaching was a serious problem on that one as well
when they went up to high zooms.
The STED module works only for one color (far red) and does not work
well with all fluorophores (--specifically, Cy5 apparently bleaches too
fast to be useful). The fluorophores they recommended are the ATTO 647
and 655 dyes. Although STED provides an improvement in x-y resolution,
there's little or no improvement in the z-axis resolution.
The instrument is essentially a Leica multiphoton microscope with the
STED unit as an attachment. It can be used in single-photon,
multiphoton or STED modes. Price is $1.3 million USD.
I'd be interested in hearing from other folks who talked to Leica about
this machine and who saw one of the demos. If I were buying one of
these items, it'd be worried about it suddenly becoming obsolete (as
happened with some of the early 2-photon instruments) due to some new
development in the pipeline. Does anyone have any sense of how likely
that is? I'd also be concerned about how suited it is to all
preparations--will it work only with the strongest labeling? Do the
ATTO dyes require particular mounting media?
Thanks in advance!
Martin
--
Martin Wessendorf, Ph.D. office: (612) 626-0145
Assoc Prof, Dept Neuroscience lab: (612) 624-2991
University of Minnesota Preferred FAX: (612) 624-8118
6-145 Jackson Hall, 321 Church St. SE Dept Fax: (612) 626-5009
Minneapolis, MN 55455 E-mail: martinw[at]med.umn.edu
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