Oct 12, 2022

Public workspaceSnap-Frozen Tissue Preparation

DOI
https://dx.doi.org/10.17504/protocols.io.n92ldzoyxv5b/v1
  • Stephen Fisher1,
  • Marielena Grijalva1,
  • Rong Guo1,
  • sarahjoh 1,
  • Hieu Nguyen1,
  • John Renz2,
  • Jean G Rosario1,
  • Steven Rudich2,
  • Brian Gregory1,
  • Junhyong Kim1,
  • Kate O'Neill1
  • 1University of Pennsylvania;
  • 2Gift of Life Donor Program
Stephen Fisher
Icon indicating open access to content
QR code linking to this content
DOI: https://dx.doi.org/10.17504/protocols.io.n92ldzoyxv5b/v1
Protocol Citation: Stephen Fisher, Marielena Grijalva, Rong Guo, sarahjoh , Hieu Nguyen, John Renz, Jean G Rosario, Steven Rudich, Brian Gregory, Junhyong Kim, Kate O'Neill 2022. Snap-Frozen Tissue Preparation. protocols.io https://dx.doi.org/10.17504/protocols.io.n92ldzoyxv5b/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 14, 2022
Last Modified: October 12, 2022
Protocol Integer ID: 68611
Keywords: biobanking of fresh frozen tissue, frozen tissue preparation this protocol, frozen tissue preparation, fresh frozen tissue, frozen tissue, preservation of tissue, snap freezing, stable in nonfixed surgical specimen, nci biospecimen evidence, intact tissue morphology, freezing protocol, nonfixed surgical specimen, freezing, national cancer institute biospecimen evidence, biobanking, placing tissue, dry ice, tissue, ideal for downstream assay, biopreservation, downstream assay, biospecimen, preservation, liquid nitrogen, rna, tissue preparation
Funders Acknowledgements:
NIH
Grant ID: U54HD104392
Abstract
This protocol describes preservation of tissue by snap-freezing. Biospecimens preserved with this protocol will be suitable for downstream analysis of DNA, RNA, protein, and morphology endpoints. Multiple workflows can be used for snap freezing including using placing tissue in a vessel either with or without media and exposing it to liquid nitrogen vapor, immersing it in liquid nitrogen, or placing it on dry ice with a cooling device (Micke et al., 2006). The workflow pursued depends on the desired balance between feasibility, time, and cost as well as the possible downstream analyses performed. The snap-freezing protocol detailed below may not be ideal for downstream assays that require intact tissue morphology (Engel, Vaught, & Moore, 2014) (see NCI Biospecimen Evidence-Based Practices).

Micke, P., Ohshima, M., Tahmasebpoor, S., Ren, Z.-P., Ostman, A., Pontén, F., & Botling, J. (2006). Biobanking of fresh frozen tissue: RNA is stable in nonfixed surgical specimens.Laboratory Investigation; a Journal of Technical Methods and Pathology,86(2), 202–211. https://doi.org/10.1038/labinvest.3700372

Engel, K. B., Vaught, J., & Moore, H. M. (2014). National Cancer Institute Biospecimen Evidence-Based Practices: A novel approach to pre-analytical standardization.Biopreservation and Biobanking,12(2), 148–150. https://doi.org/10.1089/bio.2013.0091
Materials
  • Freshly dissected tissue block
  • ReagentDry Ice
  • Reagent1.5 mL LoBind tubes EppendorfCatalog #022431021
  • Access to -80°C ultra-low freezer


Troubleshooting
Place pre-weighed piece of tissue in either a 1.5mL Eppendorf tube or 1.5mL cryo tube.
Quick freeze by immediately placing tube on dry ice.
Transfer tube to a -80℃ ultra-low freezer for Biobanking or until ready for downstream processing.