Tissue block staining and domestic adhesive tape yield qualified integral sections of adult mouse orbits and eyeballs.

Autor: Li Z; Advancecor GmbH, Martinsried, Germany., Ungerer M; ISAR Bioscience Institute, Planegg, Germany., Faßbender J; Advancecor GmbH, Martinsried, Germany., Wenhart C; Advancecor GmbH, Martinsried, Germany., Holthoff HP; Advancecor GmbH, Martinsried, Germany., Muench G; Advancecor GmbH, Martinsried, Germany.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2021 Aug 04; Vol. 16 (8), pp. e0255363. Date of Electronic Publication: 2021 Aug 04 (Print Publication: 2021).
DOI: 10.1371/journal.pone.0255363
Abstrakt: The standard histological processing procedure, which produces excellent staining of sections for most tissues, fails to yield satisfactory results in adult mouse orbits or eyeballs. Here, we show that a protocol using tissue block staining and domestic adhesive tapes resulted in qualified integral serial cryo-sections of whole orbits or eyeballs, and the fine structures were well preserved. The histological processing protocol comprises paraformaldehyde fixation, ethylenediaminetetraacetic acid decalcification, tissue block staining with hematoxylin and eosin, embedding, adhesive tape aided sectioning, and water-soluble mounting. This protocol was proved to be the best in comparison with seven other related existing histological traditional or non-traditional processing methods, according to the staining slice quality. We observed a hundred percent success rate in sectioning, collection, and mounting with this method. The reproducibility tested on qualified section success rates and slice quality scores confirmed that the technique is reliable. The feasibility of the method to detect target molecules in orbits was verified by successful trial tests on block immunostaining and adhesive tape-aided sectioning. Application of this protocol in joints, brains, and so on,-the challenging integral sectioning tissues, also generated high-quality histological staining sections.
Competing Interests: The authors have read the journal’s policy and have the following competing interests: Julia Faßbender, Clara Wenhart, Hans-Peter Holthoff, and Goetz Muench are paid employees of Advancecor GmbH of Germany. There are no patents, products in development or marketed products associated with this research to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials.
Databáze: MEDLINE
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