Confocal Microscopy & Flow Cytometry

Confocal Microscopy

confocal NSTORM A confocal fluorescence microscope is much the same as a conventional light microscope with added features to enhance its capabilities.

  • The conventional microscope uses visible light (400-700 nanometers) to illuminate and produce a magnified image of a sample.
  • A fluorescence microscope, on the other hand, uses a much higher intensity light source which excites a fluorescent species in a sample of interest. This fluorescent species in turn emits a lower energy light of a longer wavelength that produces the magnified image instead of the original light source.

Fluorescent microscopy is often used to image specific features of small specimens such as microbes.confocalWorking It is also used to visually enhance 3-D features at small scales. This can be accomplished by attaching fluorescent tags to anti-bodies that in turn attach to targeted features, or by staining in a less specific manner. When the reflected light and background fluorescence is filtered in this type of microscopy the targeted parts of a given sample can be imaged. This gives an investigator the ability to visualize desired organelles or unique surface features of a sample of interest.

Flow Cytometry

Flow cytometry is the measurement of cells in a flow system, which delivers the cells singly past a point of measurement which is illuminated by a light source.  The scattered light and fluorescence of different wavelengths are then recorded. Typically, light scattered at two different angles and from one to six or more fluorescences will be measured.

Fluorescent chemicals may used to label cell components, such as DNA, directly; others are attached to antibodies against a wide variety of cellular proteins.

The value of the technique lies in the ability to make measurements on large numbers of single cells within a short period of time (tens of seconds to minutes). The heterogeneity of populations can be revealed and different subsets of cells identified and quantified.


 

These facilities comprise the Confocal Microscopy Suite located in the Philip Lyle Building...

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