Re: imaging spheroids

Posted by Pamela Young on
URL: http://confocal-microscopy-list.275.s1.nabble.com/imaging-spheroids-tp7581903p7582145.html

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We¹ve also been having this exact problem (and on our Leica SP5).  The 20x
0.7 multi-immersion lens doesn¹t have a long enough working distance.  We
have been using a 20x 0.5 lens with a uge working distance, but we can¹t
get the spheroids to sit down in the bottom of the collagen gel, so even
it doesn¹t get us all of the way through.  I¹d love to try out a 20x/0.95
lens with the 2.5mm working distance, but we don¹t have one.  I¹ve already
brought up the light sheet idea, especially since another group on campus
is getting one.  I really think that is the best thing for spheroids in
collagen gels.  In the mean time, if anyone has spec prep thoughts for
getting spheroids closer to the coverslip when in collagen gels, I¹d love
thoughts.  

THANKS!

Dr Pamela A. Young
 | Light and Optical Microscopist
Australian Centre for Microscopy & Microanalysis

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On 11/04/2014 4:12 am, "Peter Gabriel Pitrone" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com and include the link in your posting.
>*****
>
>Beautiful data!!
>
>--
>Peter Gabriel Pitrone - FRMS TechRMS
>Microscopy/Imaging Specialist
>Prof. Dr. Pavel Tomancak group
>Max Planck Institute for
>Molecular Biology and Genetics
>Pfotenhauerstr. 108
>01307 Dresden
>
>http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html &
>http://www.openspim.org
>
>"If a straight line fit is required, obtain only two data points." - Anon.
>
>
>On Thu, April 10, 2014 19:53, jens rietdorf wrote:
><|> *****
><|> To join, leave or search the confocal microscopy listserv, go to:
><|> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
><|> Post images on http://www.imgur.com and include the link in your
>posting.
><|> *****
><|>
><|> FYI recent publication of lightsheet microscopy of tumor spheroids.
><|>
>http://www.researchgate.net/publication/51862390_Live_cell_division_dynami
>cs_monitoring_in_3D_large_spheroid_tumor_models_using_light_sheet_microsco
>py
><|> https://www.youtube.com/watch?v=72OPWhC-zy4
><|> Cheers, jens
><|>
><|>
><|> On Thu, Apr 10, 2014 at 6:40 PM, Peter Gabriel Pitrone
><|> <[hidden email]>wrote:
><|>
><|>> *****
><|>> To join, leave or search the confocal microscopy listserv, go to:
><|>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
><|>> Post images on http://www.imgur.com and include the link in your
><|>> posting.
><|>> *****
><|>>
><|>> Hello Doug,
><|>>
><|>> I agree with Diaspro (even though he didn't come out and directly say
><|>> it), Lightsheet microscopy would be better in this particular
><|>> experiment.
><|>> I'll make a shameless attempt at self promotion as well, check out
><|>> OpenSPIM: http://www.openspim.org ;-)
><|>>
><|>> Best Regards,
><|>> Pete
><|>>
><|>> --
><|>> Peter Gabriel Pitrone - FRMS TechRMS
><|>> Microscopy/Imaging Specialist
><|>> Prof. Dr. Pavel Tomancak group
><|>> Max Planck Institute for
><|>> Molecular Biology and Genetics
><|>> Pfotenhauerstr. 108
><|>> 01307 Dresden
><|>>
><|>> http://www.mpi-cbg.de/research/research-groups/pavel-tomancak.html &
><|>> http://www.openspim.org
><|>>
><|>> "If a straight line fit is required, obtain only two data points." -
><|>> Anon.
><|>>
><|>>
><|>> On Thu, April 10, 2014 17:43, Alberto Diaspro wrote:
><|>> <|> *****
><|>> <|> To join, leave or search the confocal microscopy listserv, go to:
><|>> <|> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
><|>> <|> Post images on http://www.imgur.com and include the link in your
><|>> posting.
><|>> <|> *****
><|>> <|>
><|>> <|> We got some super resolution on spheroids
><|>> <|> http://www.nature.com/nmeth/journal/v8/n12/full/nmeth.1744.html
><|>> <|>
><|>> <|> alby
><|>> <|>
><|>> <|> www.nic.iit.it
><|>> <|>
><|>> <|>
><|>> <|> Il giorno 10 apr, 2014, alle ore 17:38, Cromey, Douglas W -
><|>> (dcromey)
><|>> <|> <[hidden email]> ha scritto:
><|>> <|>
><|>> <|>> *****
><|>> <|>> To join, leave or search the confocal microscopy listserv, go
>to:
><|>> <|>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
><|>> <|>> Post images on http://www.imgur.com and include the link in your
><|>> <|>> posting.
><|>> <|>> *****
><|>> <|>>
><|>> <|>> I am working with a lab that is interested in doing fluorescence
><|>> <|>> microscopy on spheroid assays (clumps of cancer cells seeded and
><|>> <|>> growing inside a moderately thick collagen matrix).  They are
><|>> looking
><|>> <|>> at a number of different microscopy techniques on campus.
>Because
><|>> our
><|>> <|>> Leica confocal was mostly configured for 2D cultured cell and
><|>> tissue
><|>> <|>> sections, we have quickly discovered the working distance
><|>> limitations
><|>> <|>> of our existing objectives.  Our local Leica technical
><|>> representative
><|>> <|>> will be loaning us Leica's fabulous 25x/0.95 water immersion
><|>> objective
><|>> <|>> (2.5mm WD) to try out.  This should help with WD issues, as well
><|>> as
><|>> <|>> spherical aberration in the sample (their dishes do at least
>have
><|>> #1.5
><|>> <|>> thickness glass coverslip bottoms).
><|>> <|>>
><|>> <|>> Has anyone else worked with these types of assays before?  Any
><|>> <|>> suggestions on the sample prep side or the imaging side to end
>up
><|>> with
><|>> <|>> better image data?
><|>> <|>>
><|>> <|>> Thanks,
><|>> <|>> Doug
><|>> <|>>
><|>> <|>> ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
><|>> <|>> Douglas W. Cromey, M.S. - Associate Scientific Investigator
><|>> <|>> Dept. of Cellular & Molecular Medicine, University of Arizona
><|>> <|>> 1501 N. Campbell Ave, Tucson, AZ  85724-5044 USA
><|>> <|>>
><|>> <|>> office:  AHSC 4212         email: [hidden email]
><|>> <|>> voice:  520-626-2824       fax:  520-626-2097
><|>> <|>>
><|>> <|>> http://swehsc.pharmacy.arizona.edu/micro
><|>> <|>> Home of: "Microscopy and Imaging Resources on the WWW"
><|>> <|>>
><|>> <|>> UA Microscopy Alliance -
><|>> <|>> http://microscopy.arizona.edu<http://microscopy.arizona.edu/>
><|>> <|>
><|>>
><|>