A simple and fast method for tissue cryohomogenization enabling multifarious molecular extraction

• Multiple distinct molecular classes are often desired from a given tissue sample.
• We describe a simple method for cryohomogenization of tissue to enable this.
• The method yields RNA and protein with typical abundance and high quality.
• The method minimizes the number of animals required for molecular neuroscience.
• Cryohomogenization may be especially useful for lateralized brain structures.

Research in molecular biology often relies on parallel analysis of nucleic acids, protein and other molecules from a given tissue. When extracted from a single sample however, the quality and quantity of these products can be compromised. One solution is to obtain near-identical samples from multiple animals and dedicate each to a given molecular component, but this approach is not optimal from both an operational and ethical perspective. Thus, we refined the methods for cryohomogenization to allow efficient use of a single experimental sample so that it can easily be divided into fractions for extraction of different molecular components, immediately or after storage. Using western blot, nanodrop UV/V spectrometry, and a bioanalyzer, we show that cryohomogenized hippocampus samples provide high-quality RNA and protein without significant loss in abundance. The method may be particularly advantageous for parallel molecular extraction from brain structures with known hemispheric lateralization, such as the hippocampus, parietal cortex, suprachiasmatic nucleus, and amygdala.

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