bubbling media and imaging
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Hi folks, I in the middle of a bunch of experiments where I
need to bubble gas into media while imaging (Going from normoxic to hypoxic media).
Obviously the bubbles perturb the quality of the DIC image. I have tried all
sorts of homemade diffusers to minimize the effect but to no avail... So has
anyone of you developed a solution to this problem? I could exchange the
media, however regassing happens really quickly and the effects we are
measuring are subtle. Thus any changes in ionic concentrations or temperature
may lead to a similar effect. Ideas anyone simon Simon C. Watkins Ph.D, FRCPath Professor and Vice Chair, Cell Biology and Physiology Professor, Immunology Director, Center for Biologic Imaging BSTS 225, University of Pittsburgh 3500 Terrace St. Pittsburgh PA 15261 Tel: 412-352-2277 Fax:412-648-2797 URL: http://www.cbi.pitt.edu |
 
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Rietdorf, Jens |
Re: bubbling media and imaging
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear Simon, there are several transparent plastics
which are permeable for gases. If the volume you image is really small and the
surface is really large, you might be able to avoid the bubbles by blowing oxygen
on the chamber to change the properties of the fluid inside. Ibidi.de (no
commercial interest) build chambers from such materials [these plastics can also
be used for DIC imaging], also ivss.de (again no commercial interest) use gas
permeable plastics. Good luck, jens From: Confocal Microscopy
List [mailto:[hidden email]] On Behalf Of Watkins, Simon
C Search
the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Hi folks, I in the middle of a bunch of experiments where I
need to bubble gas into media while imaging (Going from normoxic to hypoxic
media). Obviously the bubbles perturb the quality of the DIC image.
I have tried all sorts of homemade diffusers to minimize the effect but to no
avail... So has anyone of you developed a solution to this problem? I
could exchange the media, however regassing happens really quickly and the
effects we are measuring are subtle. Thus any changes in ionic
concentrations or temperature may lead to a similar effect. Ideas anyone simon Simon C. Watkins Ph.D, FRCPath Professor and Vice Chair, Cell Biology and Physiology Professor, Immunology Director, Center for Biologic Imaging BSTS 225, University of Pittsburgh 3500 Terrace St. Pittsburgh PA 15261 Tel: 412-352-2277 Fax:412-648-2797 URL: http://www.cbi.pitt.edu |
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In reply to this post by Watkins, Simon C
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear Simon,
Can you pre-bubble a reservoir and then pump (or gravity feed) this into your specimen bath?
Cheers Steve Stephen H. Cody Tip: Learn how to receive reminders about you microscope
booking: -----Original Message-----
Hi folks, I in the middle of a bunch of experiments where I need to bubble gas into media while imaging (Going from normoxic to hypoxic media). Obviously the bubbles perturb the quality of the DIC image. I have tried all sorts of homemade diffusers to minimize the effect but to no avail... So has anyone of you developed a solution to this problem? I could exchange the media, however regassing happens really quickly and the effects we are measuring are subtle. Thus any changes in ionic concentrations or temperature may lead to a similar effect. Ideas anyone simon
Simon C. Watkins Ph.D, FRCPath Professor and Vice Chair, Cell Biology and Physiology Professor, Immunology Director, Center for Biologic Imaging BSTS 225, University of Pittsburgh 3500 Terrace St. Pittsburgh PA 15261 Tel: 412-352-2277 Fax:412-648-2797 URL: http://www.cbi.pitt.edu
This communication is intended only for the named recipient and may contain information that is confidential, legally privileged or subject to copyright; the Ludwig Institute for Cancer Research does not waiver any rights if you have received this communication in error. The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research. |
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In reply to this post by Watkins, Simon C
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Simon I would propose to use a closed perfusion chamber (yes, we sell one, so there's some commercial interest here) and perfuse it with pre-equilibrated medium. Since there is no fluid-gas interface in the chamber, there will be no optical perturbation due to "ripples" on the surface either. There are some other things to consider, so feel free to contact me off the list. Beat At 15:13 28-07-2008, you wrote: >Search the CONFOCAL archive at >http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal >Hi folks, I in the middle of a bunch of experiments where I need to >bubble gas into media while imaging (Going from normoxic to hypoxic >media). Obviously the bubbles perturb the quality of the DIC >image. I have tried all sorts of homemade diffusers to minimize the >effect but to no avail... So has anyone of you developed a solution >to this problem? I could exchange the media, however regassing >happens really quickly and the effects we are measuring are >subtle. Thus any changes in ionic concentrations or temperature may >lead to a similar effect. >Ideas anyone >simon > >Simon C. Watkins Ph.D, FRCPath >Professor and Vice Chair, Cell Biology and Physiology >Professor, Immunology >Director, Center for Biologic Imaging >BSTS 225, University of Pittsburgh >3500 Terrace St. >Pittsburgh PA 15261 >Tel: 412-352-2277 >Fax:412-648-2797 >URL: http://www.cbi.pitt.edu > |
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In reply to this post by Watkins, Simon C
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Simon, If you are just after a hypoxic media, it should be easiest and best to work under an atmosphere of nitrogen, rather than bubbling your samples - the concentration in your solution will depend on the solubility of oxygen in the media and the oxygen concentration in the atmosphere (Henry's law). As far as I know you will not be able to remove 100% oxygen from aqueous solutions anyway (without freezing and/or high vacuum), so you may as well work with nearly oxygen free media. We use a mini-incubation chamber, sold by lots of suppliers, but a petidish with holes for gas inlet and outlet could also be sufficient. Sonja Citat af "Watkins, Simon C" <[hidden email]>: > Search the CONFOCAL archive at > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal > > Hi folks, I in the middle of a bunch of experiments where I need to > bubble gas into media while imaging (Going from normoxic to hypoxic > media). Obviously the bubbles perturb the quality of the DIC image. > I have tried all sorts of homemade diffusers to minimize the > effect but to no avail... So has anyone of you developed a solution > to this problem? I could exchange the media, however regassing > happens really quickly and the effects we are measuring are subtle. > Thus any changes in ionic concentrations or temperature may lead > to a similar effect. > Ideas anyone > simon > > Simon C. Watkins Ph.D, FRCPath > Professor and Vice Chair, Cell Biology and Physiology > Professor, Immunology > Director, Center for Biologic Imaging > BSTS 225, University of Pittsburgh > 3500 Terrace St. > Pittsburgh PA 15261 > Tel: 412-352-2277 > Fax:412-648-2797 > URL: http://www.cbi.pitt.edu > > Sonja Hatz Department of Chemistry University of Aarhus Langelandsgade 140 DK-8000 Aarhus C Denmark Tel: (45) 8942 3860 Fax: (45) 8619 6199 ---------------------------------------------------------------- This message was sent using IMP, the Internet Messaging Program. |
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