Oct 04, 2023
UPitt TriState SenNet TMC Cryopreserved Tissue Grinding
- 1Division of Pulmonary and Critical Care Medicine. Department of Medicine. School of Medicine
Protocol Citation: Marta Bueno 2023. UPitt TriState SenNet TMC Cryopreserved Tissue Grinding. protocols.io https://dx.doi.org/10.17504/protocols.io.q26g7px41gwz/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 04, 2023
Last Modified: October 04, 2023
Protocol Integer ID: 88807
Funders Acknowledgement:
TriState SenNET (Lung and Heart) Tissue Map and Atlas consortium - NIA
Grant ID: U54AG075931
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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).
Guidelines
Ensure the samples are kept frozen on dry ice throughout pulverization.
Safety warnings
Use universal safety precautions when handling human samples and personal protective equipment (e.g., face mask with shield, gloves, lab coat or apron).
Before start
- Use clean and/or autoclaved tools.
- Use RNAZap over all sufaces and be aware of cross-contamination sources.
Tissue pulverization
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.