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.

Mascadri, F., Bolognesi, M., Pilla, D., Cattoretti, G. (2021). Rejuvenated Vintage Tissue Sections Highlight Individual Antigen Fate During Processing and Long-term Storage. JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 69(10), 659-667 [10.1369/00221554211047287].

Rejuvenated Vintage Tissue Sections Highlight Individual Antigen Fate During Processing and Long-term Storage

Bolognesi, Maddalena M
Co-primo
Membro del Collaboration Group
;
Cattoretti, Giorgio
Ultimo
Membro del Collaboration Group
2021

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.
Articolo in rivista - Articolo scientifico
antigen retrieval; antigenic variation; fluorescent image analysis; formalin-fixed paraffin-embedded tissue section; immunohistochemistry; long-term storage of FFPE tissue section; quantitative methods
English
19-set-2021
2021
69
10
659
667
none
Mascadri, F., Bolognesi, M., Pilla, D., Cattoretti, G. (2021). Rejuvenated Vintage Tissue Sections Highlight Individual Antigen Fate During Processing and Long-term Storage. JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 69(10), 659-667 [10.1369/00221554211047287].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/327763
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