Vibratome
A vibratome being used to slice a rat brain.
A vibratome is an instrument that is similar to a microtome but uses a vibrating razor 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 glued 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]
Advantages of Vibratome Sectioning
- No need to dehydrate tissues prior to embedding, thus decreased loss of cell constituents
- No messy paraffin embedding
- 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 parrafin embedding or freezing
- Increased tissue autofluoresecence due to paraffin embedding being avoided
- Less wait period from tissue sampling to time of immunolabelling
Possible Disadvantages of Vibratome Sectioning
- 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
- ↑ http://www.vibratome.com/global/Manuals/VB100AppNote1.PDF
- ↑ http://www4.ncsu.edu/~rgfranks/FRANKS%20LAB%20PROTOCOLS/immuno%20histio%20chemistry/robertson%20vbratmimuno.doc
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