Confocal scanning microscope

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roudi
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Confocal scanning microscope

A microscope that uses a Nipkow disc or a laser to produce point-probing raster scanning, yielding images with very high contrast in the third dimension. A small aperture at the secondary focus of the objective lens narrows the depth of focus and obstructs most of the light reflected from out-of-focus objects.
Can we use convocal scanning microscope to study live cells or they should be fixed?

Fraser Moss
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Live cell imaging is very

Live cell imaging is very possible. See the link below and the Imaging Modes section.

http://www.microscopyu.com/articles/confocal/

There are plenty on articles on the net if you search live cell imaging in google and in Entrez -Pubmed, or even the major microscope manufacturers (Leica, Nikon, Olympus).

If you want to do live cell imaging you may want to look into using a heated stage, and or heated perfusion systems to keep everything at physiological temperature for each experiment.

samm
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The Olympus DSU attachment is

The Olympus DSU attachment is a great tool for live cell work with minimal bleaching, provided your sections/cells are on the thicker side - going by our experience, a minimum of 20 uM is good (the instrument is capable of 100 nm sections, but you need a thick section for a proper 3D reconstruction. - Its not quite confocal, but at less than a third of the cost, and several fold less bleaching, it does a good job!
If you need to look at multiple fields, you'll (in addition to heated stage etc mentioned by frasermoss) need a fully automated stage (e.g. Prior).

MariaMaria
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samm wrote:The Olympus DSU

samm wrote:

The Olympus DSU attachment is a great tool for live cell work with minimal bleaching, provided your sections/cells are on the thicker side - going by our experience, a minimum of 20 uM is good (the instrument is capable of 100 nm sections, but you need a thick section for a proper 3D reconstruction. - Its not quite confocal, but at less than a third of the cost, and several fold less bleaching, it does a good job!
If you need to look at multiple fields, you'll (in addition to heated stage etc mentioned by frasermoss) need a fully automated stage (e.g. Prior).

Actually the website for Olympus Confocal microscopes is:
http://www.olympusamerica.com/seg_section/seg_confocal.asp

roudi
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Can I study live cells using

Can I study live cells using Bio-Rad Laser scanning Confocal Microscopy?

The BIO-RAD Radiance 2000 is a laser scanning confocal microscope system used in epifluorescence and reflected light imaging. A finely focused beam of laser light is scanned across a sample and the resultant light emitted passed through a pinhole aperture to exclude any out of focus light. The image thus produced includes only the plane of focus for a given objective lens. By varying the focussing depth a series of sequential images through the thickness of a single sample can be collected and used to project a three-dimensional image of the sample. A basic overview of the system configuration is given in the following tables