imaging spheroids

[Cromey, Douglas W - (dcromey)]

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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:  [phone]

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/>

[Alberto Diaspro]

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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:  [phone]
>
> 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/>

[Feinstein, Timothy]

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Hi Doug,

The short answer is to use a water immersion lens on an upright scope.
However, Leica makes a 20x NA 0.7 multi-immersion objective that you
should check out (no commercial interest).  I have used both the Nikon and
Zeiss equivalents on inverted scopes when we needed max imaging depth.

The deeper you image the more you will lose to spherical aberration unless
you match the refractive index of lens immersion with your sample medium.
A multi-immersion lens lets you do that better than most alternatives.  On
the other hand flexibility always necessitates optical trade-offs; others
here can describe the considerations there better than I can.

All the best,


TF

Timothy Feinstein, Ph.D. | Confocal Manager
Van Andel Research Institute
333 Bostwick Ave., N.E., Grand Rapids, Michigan 49503
Phone: [phone] | Email: [hidden email]







On 4/10/14, 11:38 AM, "Cromey, Douglas W - (dcromey)"
<[hidden email]> wrote:

>*****
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>HSH5pdA&u=http%3a%2f%2flists%2eumn%2eedu%2fcgi-bin%2fwa%3fA0%3dconfocalmic
>roscopy
>Post images on
>http://scanmail.trustwave.com/?c=129&d=hrvG04ykaG4omxGFWfNLSjePzRtNR9vkPXV
>GGnQ9JA&u=http%3a%2f%2fwww%2eimgur%2ecom 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:  [phone]
>
>http://scanmail.trustwave.com/?c=129&d=hrvG04ykaG4omxGFWfNLSjePzRtNR9vkPXU
>SHHJkJg&u=http%3a%2f%2fswehsc%2epharmacy%2earizona%2eedu%2fmicro
>Home of: "Microscopy and Imaging Resources on the WWW"
>
>UA Microscopy Alliance -
>http://scanmail.trustwave.com/?c=129&d=hrvG04ykaG4omxGFWfNLSjePzRtNR9vkPSM
>XE3JuIQ&u=http%3a%2f%2fmicroscopy%2earizona%2eedu<http://scanmail.trustwav
>e.com/?c=129&d=hrvG04ykaG4omxGFWfNLSjePzRtNR9vkPXEVGH4-Jw&u=http%3a%2f%2fm
>icroscopy%2earizona%2eedu%2f>

[Peter Gabriel Pitrone]

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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:  [phone]
<|>>
<|>> 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/>
<|>

[jerie]

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*****

FYI recent publication of lightsheet microscopy of tumor spheroids.
http://www.researchgate.net/publication/51862390_Live_cell_division_dynamics_monitoring_in_3D_large_spheroid_tumor_models_using_light_sheet_microscopy
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:  [phone]
> <|>>
> <|>> 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/>
> <|>
>

[Reto Fiolka]

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*****

Hi all,
As a light sheet microscope's detection is widefield, the overall penetration depth
of the system will be limited to less than one scattering mean free path (~in the
order of 100 microns for most tissues). This might be reached in the Zeiss
youtube example and hence things get very blurry.

A two-photon scanning microscope can go up to 5 scattering path lengths, the
strehl ratio will be much reduced, but you can still see cells.

Best,
Reto

[Peter Gabriel Pitrone]

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*****

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_dynamics_monitoring_in_3D_large_spheroid_tumor_models_using_light_sheet_microscopy
<|> 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:  [phone]
<|>> <|>>
<|>> <|>> 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/>
<|>> <|>
<|>>
<|>

[Pamela Young]

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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:  [phone]
><|>> <|>>
><|>> <|>> 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/>
><|>> <|>
><|>>
><|>