http://confocal-microscopy-list.275.s1.nabble.com/HCS-in-vivo-imaging-system-tp591433p591435.html
expressing cells in liver and lymphnodes. What is your
Fox. Do you know any users who have sucessfuly used
Once again thanks a lot for the information.
> very good.
>
> 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.asp>
> If 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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