Aug 30, 2024

Public workspaceKAPP-Sen TMC: Tissue blocks paraffin embedding by JAX Histology Services (RNase-free)

DOI
dx.doi.org/10.17504/protocols.io.rm7vzj9z2lx1/v1
  • Anne Marchini1,
  • Tim Adams1,
  • Elaine Bechtel1,
  • Lesley Bechtold1
  • 1The Jackson Laboratory, Bar Harbor, ME, USA
  • Cellular Senescence Network (SenNet) Method Development Community
  • KAPP-Sen TMC
Sergii Domanskyi
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DOI: dx.doi.org/10.17504/protocols.io.rm7vzj9z2lx1/v1
Protocol Citation: Anne Marchini, Tim Adams, Elaine Bechtel, Lesley Bechtold 2024. KAPP-Sen TMC: Tissue blocks paraffin embedding by JAX Histology Services (RNase-free). protocols.io https://dx.doi.org/10.17504/protocols.io.rm7vzj9z2lx1/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: August 27, 2024
Last Modified: August 30, 2024
Protocol Integer ID: 106502
Keywords: KAPP-Sen TMC
Funders Acknowledgement:
National Institute on Aging (NIA) KAPP-Sen Tissue Mapping Collaborative
Grant ID: U54 AG075941
Abstract
This protocol describes embedding of formalin-fixed tissues in paraffin (FFPE). Paraffin-embedded blocks are then kept at 4 °C and protected from light until RNA quality and spatial omics assays are performed.
Wear prescribed PPE throughout all of the procedures described in this document. Change gloves if they get wet.
Histology does not process tissue by hand for RNase free requests. The usual tissue processing unit (Leica Histocore Peloris 3) is used. Use RNase free best practices when handling the tissue (wear gloves that have been wiped with RNAse Zap wipes) to load the tissue onto the processor and then into the embedding station.
Place cassettes containing tissue samples into a clean and empty paraffin bath within the embedding unit (Sakura Tissue-Tek TEC5 Embedding Console System). To avoid contamination, do not immerse in liquid paraffin within the embedding center.
Wipe gloves, molds and forceps with RNase Zap wipes before use. Allow the molds and forceps to dry completely. Use a pair of clean forceps for each tissue type. Do not re-use molds without cleaning between samples.
After the blocks are cooled and removed from the embedding molds, don gloves cleaned with RNase Zap wipes. Then place the blocks in a plastic bag labeled with the job number and store them in a 4 °C refrigerator protected from light until they are sectioned.
Store paraffin blocks in a 4 °C refrigerator prior to cutting; RNA continues to degrade.