Evaluation of Long-Term Cryostorage of Brain Tissue Sections for Quantitative Histochemistry
| Autor: | Nadine C. Heyworth, Howard Cabral, Douglas L. Rosene, Larissa I. Estrada, Laura B. Ngwenya, Eustathia Lela Giannaris, Amy A. Robinson, Ana C. Amaral, Farzad Mortazavi, Ronald J. Killiany, Debra Roberts |
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| Rok vydání: | 2017 |
| Předmět: |
Doublecortin Domain Proteins
Male 0301 basic medicine Pathology medicine.medical_specialty Histology Cell Count Nerve Tissue Proteins Stereology Cryopreservation 03 medical and health sciences 0302 clinical medicine medicine Animals Frozen Sections Hyaluronic Acid Brain Chemistry Orexins Frozen section procedure biology Chemistry Brain-Derived Neurotrophic Factor Neuropeptides Antigens Nuclear Myelin Basic Protein Articles Immunohistochemistry Macaca mulatta Doublecortin Myelin basic protein Parvalbumins 030104 developmental biology Bromodeoxyuridine biology.protein Female Anatomy Densitometry Microtubule-Associated Proteins 030217 neurology & neurosurgery Parvalbumin |
| Zdroj: | Journal of Histochemistry & Cytochemistry. 65:153-171 |
| ISSN: | 1551-5044 0022-1554 |
| Popis: | Storage of tissue sections for long periods allows multiple samples, acquired over months or years, to be processed together, in the same reagents, for quantitative histochemical studies. Protocols for freezer storage of free-floating frozen sections using sucrose with different additives have been reported and assert that storage has no effect on histochemistry, but no quantitative support has been provided. The present study analyzed the efficacy of long-term storage of brain tissue sections at −80C in buffered 15% glycerol. To determine whether histochemical reactivity is affected, we analyzed 11 datasets from 80 monkey brains that had sections stored for up to 10 years. For processing, sections from multiple cases were removed from storage, thawed, and batch-processed at the same time for different histochemical measures, including IHC for neuronal nuclear antigen, parvalbumin, orexin-A, doublecortin, bromodeoxyuridine, the pro-form of brain-derived neurotrophic factor, and damaged myelin basic protein as well as a histochemical assay for hyaluronic acid. Results were quantified using stereology, optical densitometry, fluorescence intensity, or percent area stained. Multiple regression analyses controlling for age and sex demonstrated the general stability of these antigens for up to a decade when stored in 15% glycerol at −80C. |
| Databáze: | OpenAIRE |
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