Posted by
George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/HCS-in-vivo-imaging-system-tp591433p591434.html
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocalHi Nishigandha,
As always - get a demo! Really more a question of budget and throughput.
For in vivo, the Caliper/Xenogen IVIS Spectrum looks very good.
http://www.caliperls.com/products/optical-imaging/ivis-spectrum.htm
My colleagues at UM are upgrading our IVIS 200 to the Spectrum in a
month. Advantage of the IVIS systems is that using cryogenic
back-illuminated CCD's enables luciferase bioluminescence imaging
(BLI) in vivo (and ex vivo - extremely valuable to take out the
organs at the end and re-image without the rest of the animal in the
way. also, can image multi-well plates, so can test the performance
of cell lines before putting them in the mouse).
A huge advantage of the Xenogen systems is that they have made the
effort to provide quantitative data for bioluminescence. They use
photons/second/cm^2/steradian. This enables every paper published
with a Xenogen system to be compared to each other (of course, the
depth of the cells, expression level, and amount of luciferin at the
cells are additional variables). If you get a non-Xenogen BLI
instrument, make sure the vendor validates the same units. Xenogen
now does something similar for fluorescence tomography (a trickier issue).
CRi has been in the in vivo fluorescence field for several years.
Check out their
http://www.cri-inc.com/products/maestro2.aspIf your budget is on the low end, arrange a demo from John Fox of
Lightools (
http://www.lightools.com/tutorial.htm). I was especially
impressed by a demo of his a couple of years ago, that included the
Pan-a-See-Ya (
http://www.lightools.com/LRTPDFS/panaseeypdf.pdf), a
simple RGB color camera, and mouse in hand without anesthesia (you
need an isofluorene rig for the other systems).
Reagents, cells and animals for in vivo imaging:
Marker Gene Technology and Promega both sell gal-luciferin, a LacZ
substrate whose product is a firefly luciferase substrate. Sam
Gambhir has lots of tribrid vectors of FP-luciferase-TK (TK = enzyme
for use with PET reporter and therapy).
Xenogen also has several transgenic mice and various cell lines
expressing luciferase. GFP (and maybe by now RFP) and firefly
luciferase transgenic mice are available from Jackson Labs
(www.jax.org). Lots of plasmids are available from www.addgene.com
Which FP works best is tissue dependent - see
http://www.jhc.org/cgi/content/abstract/55/9/931 for one study.
You can also demo by injecting known volumes of fluorescent beads
(i.e. in matrigel) under the skin, though you should expect most of
the research to be of deeper sites. Ralph Weissleder's company, Visen
Medical, sells a variety of fluorescence reagents for mouse in vivo
imaging (AngioSense 680, etc). Fluorescence angiography with
indocyanine green (ICG) or fluorescein
(
http://msp.rmit.edu.au/Article_02/03d.html) has been clinically
available for years, and is a good way to start with mice.
HCS - Cellomics (Thermo Fisher) founded the field, worth demoing.
They had a nice article on their web site on how adding an Apotome
(Zeiss optical sectioning device) to their instrument improved
Z'-scores, so confocal imaging should help. Drug Discovery Today,
ASSAY and Drug Development Technologies, Nature Reviews Drug
Discovery, and a bunch of other journals cover this field. Start with
the following books:
Methods Enzymol. 2006;414
Methods Mol Biol. 2007;356
Then, search PubMed for high content screen*
George
At 12:56 AM 1/12/2008, you wrote:
>Search the CONFOCAL archive at
>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>
>Dear All,
>
>I am planning to procure High Content Screening system
>(HCS). I would like to know experience of users as I
>could not find many published reports on different
>systems. I am also debating on utility of confocal
>(Nipkow disk based) based HCS versus without confocal
>HCS. I will be thankful if anybody could share their
>experience or opine on the various available makes in
>the market.
>
>I am also looking for in vivo imaging system to
>procure images from small animals such as mouse, rats
>to study tumor formation, metastasis and angeogenesis
>of GFP or RFP expressing. I request to share your
>experience to enable me to choose the proper system
>for these applications.
>
>With thanks and best wishes for the year,
>
>Nishigandha Naik
>
>
>
>
>
>____________________________________________________________________________________
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http://tools.search.yahoo.com/newsearch/category.php?category=shoppingGeorge McNamara, Ph.D.
University of Miami, Miller School of Medicine
Image Core
Miami, FL 33010
[hidden email]
[hidden email]
305-243-8436 office
http://home.earthlink.net/~pubspectra/http://home.earthlink.net/~geomcnamara/http://www.sylvester.org/health_pro/shared_resources/index.asp (see
Analytical Imaging Core Facility)