Posted by
JOEL B. SHEFFIELD on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Basic-live-cell-imaging-question-tp5687698p5703919.html
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I think Gert's response makes a lot sense. For a newbie, with limited goals
(initially), it is not necessary to have an elaborate setup. In the (very)
old days, there was something called a "hanging drop culture" in which
tissue fragments were placed in a drop of medium on a slide, the slide was
inverted and examined with a conventional microscope. Gert is suggesting a
modern version of the same, assuming that you would visualize the cells
through the cover slip to which they are attached, Ujnfortunately, the
original writer did not specify the kind of optics they intended to use. If
the cells are, indeed, attached to the cover slip, it is possible to use
properly adjusted phase or Hoffman optics in transmissionto increase the
contrast. If fluorescence is needed, then you have to be concerned with
light toxicity, etc., but the optics should work fine, as long as you don't
want to see nuclear speckles, or details of mitochondria.
Joel
On Wed, Nov 3, 2010 at 9:03 PM, Axel Kurt Preuss <
[hidden email]> wrote:
> You need a water immersion object or have to build one
>
>
> Cheers
>
> Axel
> —————
> Axel K Preuss, PhD,
> Central Imaging, IMCB, A*Star, 61 Biopolis Dr, 6-19B, Singapore 138673,
> sent from 9271.5622
>
>
> On Nov 4, 2010, at 4:06 AM, Gert van Cappellen <
>
[hidden email]> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> >
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> > *****
> >
> > Culture your cells on a round coverslip. Take an object glas glue a
> > square piece of non-toxic rubber with a round hole on it. Fill this with
> > CO2 satured medium somewaht more as the volume of the hole. Put your
> > coverslip on it, with the cells to the medium off course. Press it
> > gently down and the glass will seal itself to the rubber ring. Now your
> > cells will survive for a couple of hours, so you can do the first
> > imaging. For real experiments you have to find a way to heat the object
> > glass to 37C.
> >
> > Good luck, Gert
> >
> > Op 29-10-2010 21:00, Dolphin, Colin schreef:
> >> *****
> >> To join, leave or search the confocal microscopy listserv, go to:
> >>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> >> *****
> >>
> >> We would like to do live cell imaging - mammalian cell lines - but only
> have direct access to an upright Olympus BX61. We don't really need
> complicated perfusion chambers, etc just something simple. We're real
> neophytes so all suggestions gratefully received.
> >>
> >> Colin
> >>
>
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--
Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
e-mail:
[hidden email]
URL:
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