Vibratome
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Vibratome
Vibratome produced by Leica Biosystems
 A vibratome is an instrument used to cut thin slices of material (although, usually thicker slices than those cut in paraffin-embedded samples using a microtome). It is similar to a microtome but uses a vibrating blade to cut through tissue.  The vibration amplitude, the speed, and the angle of the blade can all be controlled.  Fixed or fresh tissue pieces are embedded in low gelling temperature agarose.(Some have had success without using the agarose to embed[]) The resulting agarose block containing the tissue piece is then attached to a metal block and sectioned while submerged in a water or buffer bath.  Individual sections are then collected with a fine brush and transferred to slides or multiwell plates for staining.[1]

Vibratome is a registered trademark of Leica Biosystems Richmond, Inc. in the USA.

Advantages

  • No need to dehydrate tissues prior to embedding, thus decreased loss of cell constituents
  • No messy paraffin embedding
  • Allows for agarose embedding of tissue to provide cutting stability
  • No need to deparaffinise and rehydrate sections prior to immunostaining
  • No high temperatures or harsh chemical treatments that may lead to antigen instability
  • No special microtome blades required
  • Less chance of artifacts caused by paraffin embedding or freezing
  • Decreased tissue autofluoresecence due to avoidance of formalin-fixation and paraffin embedding
  • Less wait period from tissue sampling to time of immunolabelling
  • Allows for direct creation of free-floating sections for immunohistochemistry

Disadvantages

  • Instead of ribbons, single sections are cut and collected which are more delicate and difficult to handle.
  • Sections are generally thicker than those obtained with paraffin methods; penetration of antibodies and other reagents may be slower and thus longer incubation times may be necessary. Also, thick sections may be difficult to image with the microscope. (However, thick sections are compatible and sometimes even desirable if using confocal microscopy.)
  • Securing vibratome sections to glass slides can be difficult or impossible, due to the thickness of the sections.[2]

References

  1. ^ "Improved methods for acute brain slice preparation from adult and aging animals" (PDF). Archived from the original (PDF) on July 17, 2011. Retrieved 2009.
  2. ^ http://www4.ncsu.edu/~rgfranks/FRANKS%20LAB%20PROTOCOLS/immuno%20histio%20chemistry/robertson%20vbratmimuno.doc[permanent dead link]

  This article uses material from the Wikipedia page available here. It is released under the Creative Commons Attribution-Share-Alike License 3.0.

Vibratome
 



 



 
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