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

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...

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Published in:PloS one 2021-08, Vol.16 (8), p.e0255363-e0255363
Main Authors: Li, Zhongmin, Ungerer, Martin, Faßbender, Julia, Wenhart, Clara, Holthoff, Hans-Peter, Muench, Goetz
Format: Article
Language:English
Subjects:
Acetic acid
Acids
Adhesives
Analysis
Animal models in research
Biology and Life Sciences
Decalcification
Embedding
Ethanol
Evaluation
Experiments
Integrals
Medicine and Health Sciences
Morphology
Orbits
Physical Sciences
Physiological aspects
Research and Analysis Methods
Retina
Rodents
Sectioning
Staining
Success
Target detection
Tissues
Ultrasonic imaging
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description 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.
doi_str_mv 10.1371/journal.pone.0255363
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subjects Acetic acid
Acids
Adhesives
Analysis
Animal models in research
Biology and Life Sciences
Decalcification
Embedding
Ethanol
Evaluation
Experiments
Integrals
Medicine and Health Sciences
Morphology
Orbits
Physical Sciences
Physiological aspects
Research and Analysis Methods
Retina
Rodents
Sectioning
Staining
Success
Target detection
Tissues
Ultrasonic imaging
title Tissue block staining and domestic adhesive tape yield qualified integral sections of adult mouse orbits and eyeballs
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