Assessment of 2-Year Storage Conditions on Protein, RNA, and DNA in Unstained Human Tissue Sections, Including a Novel Multiplex Digital Gene Expression Profiling Method with Implications for Biobanking
- 1 December 2022
- journal article
- research article
- Published by Mary Ann Liebert Inc in Biopreservation and Biobanking
- Vol. 20 (6), 473-484
- https://doi.org/10.1089/bio.2021.0023
Abstract
Background: Formalin-fixed, paraffin-embedded (FFPE) tissues are a valuable resource for clinical and basic science research. Paraffin blocks and the resulting unstained sections (USS) are often stored for years before being used. Previous studies have evaluated the effects of time, temperature, humidity, and inert gases on preservation of USS; however, no study has examined all four variables together. Methods: In the current work, we prospectively and blindly assessed time points from 0 to 24 months, room versus refrigerated temperature, and presence of a desiccant and/or nitrogen atmosphere on a variety of benign and malignant tissues from North America and Africa. End points included immunohistochemistry (IHC), in situ hybridization (ISH), extracted RNA and DNA quantity and quality, and messenger RNA performance in a novel, multiplexed digital gene expression profiling assay of both housekeeping and tumor-specific genes. Results: We found that using current methods of antigen retrieval, staining, and extraction, the end points of IHC, ISH, RNA, and DNA were well preserved under the various conditions tested, with implications that pre-embedding factors contribute to variability in subsequent tissue integrity. We also document that spectrophotometric estimations of nucleic acid concentrations were in general estimated to be higher than with fluorimetric methods, which may be pertinent to end assay development. We further describe a new multiplex assay, the PlexSet digital gene expression assay, suitable for evaluating RNA quality in FFPE tissues. Conclusion: Altogether, these results may provide helpful guidance with regard to approaches for long-term storage conditions for USS.Keywords
This publication has 29 references indexed in Scilit:
- Effects of Long-Term Storage on the Detection of Proteins, DNA, and mRNA in Tissue Microarray SlidesJournal of Histochemistry & Cytochemistry, 2011
- Factors Influencing the Degradation of Archival Formalin-Fixed Paraffin-Embedded Tissue SectionsJournal of Histochemistry & Cytochemistry, 2011
- A Novel Method of Amplification of FFPET-Derived RNA Enables Accurate Disease Classification with MicroarraysThe Journal of Molecular Diagnostics, 2010
- Combined effects of formalin fixation and tissue processing on immunorecognitionBiotechnic & Histochemistry, 2009
- Combined effects of formalin fixation and tissue processing on immunorecognitionBiotechnic & Histochemistry, 2009
- Technical pitfalls potentially affecting diagnoses in immunohistochemistryJournal of Clinical Pathology, 2008
- Direct multiplexed measurement of gene expression with color-coded probe pairsNature Biotechnology, 2008
- Antigenicity testing by immunohistochemistry after tissue oxidationJournal of Clinical Pathology, 2007
- Long-term preservation of antigenicity on tissue microarraysLaboratory Investigation, 2004
- Loss of Tumor Marker-Immunostaining Intensity on Stored Paraffin Slides of Breast CancerJNCI Journal of the National Cancer Institute, 1996