1. Rejuvenated Vintage Tissue Sections Highlight Individual Antigen Fate During Processing and Long-term Storage
- Author
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Francesco Mascadri, Daniela Pilla, Giorgio Cattoretti, Maddalena Maria Bolognesi, Mascadri, F, Bolognesi, M, Pilla, D, and Cattoretti, G
- Subjects
formalin-fixed paraffin-embedded tissue section ,Tissue Fixation ,Histology ,Protein Conformation ,Reducing agent ,long-term storage of FFPE tissue section ,chemistry.chemical_element ,antigenic variation ,Calcium ,fluorescent image analysis ,Surface-Active Agents ,chemistry.chemical_compound ,Protein structure ,Antigen ,Pulmonary surfactant ,Antigenic variation ,Humans ,antigen retrieval ,quantitative methods ,fluorescent image analysi ,Antigens ,Paraffin Embedding ,Chemistry ,Proteins ,Articles ,Cell biology ,Antigen retrieval ,Tissue Array Analysis ,immunohistochemistry ,Immunohistochemistry ,Anatomy - Abstract
Antigen-bearing proteins become progressively unavailable to immunodetection after prolonged storage of routine sections, exposed to a variety of agents, such as moisture, oxygen, and temperature. By proteomic analysis, the antigens are retained in the sections and definitely in the tissue block, pointing to fixation-independent, storage time–dependent protein modifications. Based on previous experience, we hypothesized that a combined exposure to a reducing agent and to chemicals favoring protein conformation changes would reverse the masking in aged sections. Disaccharides, lactose and sucrose, and a surfactant, added to a standard antigen retrieval buffer, reverse the negative changes in aged sections. Furthermore, they provide enhanced access to antigens in freshly cut sections, but not universally, revealing additional factors, besides heat and calcium chelation, required for antigen retrieval of individual proteins
- Published
- 2021
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