George Mason University Poster Presentation for the National Cancer Institute’s 15th Annual IMAT Principle Investigators Meeting, November 13-15, 2014.
A significant and underappreciated issue is the fact that excised tissue is alive and reacting to ex vivo stress . During this “cold ischemia time” cells within the tissue react and adapt to the absence of vascular perfusion, ischemia, hypoxia, acidosis, and accumulation of cellular waste. Challenged by the realization that phosphoprotein signaling pathways were reactive and fluctuating immediately following procurement, we developed a non-formalin fixative chemistry for the preservation of biomarker molecules and histomorphology in one step, in epithelial and calcified tissues, using standard clinical pathology processing protocols  (US Patent #8,460,859).
Technology and potential advantages:
A novel, non-formalin, one-step phosphoprotein preservation chemistry was created, characterized in a wide variety of human and animal tissues, independently validated by pathologists, published, patented, licensed, and samples were distributed to the scientific community. Our preservative stabilizes phosphoproteins, immunohistochemical antigens, glycoproteins, nucleic acids, decalcifies bone, and renders exquisite diagnostic cellular histomorphology superior, or equivalent, to formalin. This fixative reduces biospecimen pre-analytical variability in research or clinical molecular profiling. Our novel preservative makes it possible to ask, and answer, research questions, and conduct clinical trial studies that were never before possible. For example, we can now assess molecular targets in calcified tissue, and reliably monitor phosphorylated signal transduction proteins – the substrates of kinase drug targets.
Progress to date:
Completed independent performance validation of the fixative in a wide variety of tissues and labile cancer related antigens. The histomorphology of our fixative was equal or superior to formalin under the following histomorphology rankings: overall color and fidelity, cell size, preservation of nuclear membrane, preservation of nucleoli and nuclear chromatin, preservation of overall cell structure, and nuclear:cytoplasmic ratio maintained. RNA quality was equal to frozen brain tissue prior to paraffin embedding, and is compatible with laser capture microdissection and qRT-PCR. In the case of calcified tissues, our fixative aided a simplified processing of bony tissues (18.5 hour reduction in processing time) while supporting immunohistology, histomorphology, and FISH, superior to or equivalent to formalin.
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