Performing an unbiased stereological study begins long before the investigator is at the microscope. The collection and preparation of sample tissue is important. Damage occurring during tissue preparation may lead to tissue that is not of the quality needed and thus great care must be taken during organ harvesting, as well as during the sectioning of tissue. During dissection care must be taken not to puncture, cut or otherwise damage the organ in question, as doing so could damage deep structures that are the target of analysis. The most likely time to damage or lose tissue is during sectioning. Before an investigator begins sectioning tissue for stereology it may be helpful to practice sectioning techniques on unimportant tissue whose loss will not affect the study.
Tips for collecting tissue:
1) Try to collect every section that comes off your cryostat, vibratome, etc… This will mean more tissue to work with later.
2) Maintain the sequential order of your tissue. Using a 96 well tissue culture plate is a great way to keep your sections in order.
3) Handle your tissue carefully. Frozen tissue is very easy to tear.
4) Use landmarks to identify the ‘handedness’ of your tissue. It may be helpful to notch the side of your organ of interest before sectioning. This will provide an unambiguous reference for left and right. This is critical when analyzing brain structures.
Sponsored by MBF Bioscience
developers of Stereo Investigator, the most cited stereology system