UPitt TriState SenNet TMC Cryopreserved Tissue Grinding
Marta Bueno
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Abstract
This document outlines the tissue pulverization protocol of lung and heart specimens at the TriState SenNet TMC Biospecimen Core at the University of Pittsburgh, as part of the Cellular Senescence Network Program (SenNet).
Before start
Use clean and/or autoclaved tools.* Use RNAZap over all sufaces and be aware of cross-contamination sources.
Steps
Tissue pulverization
Pour liquid nitrogen into a mortar and pestle until frozen.
Remove tissue from tube and place into liquid nitrogen with a clean cold spatula (precool in liquid
nitrogen).
Grind up the sample with liquid nitrogen using the mortar and pestle.
Use a clean cold spatula to scoop ground tissue to 1.5 mL LoBind tube (Eppendorf).
Place the sample back onto dry ice or directly to -80 °C.
Clean the mortar and pestle with 10% bleach and 70% ethanol before using it for the next sample.
