Detecting Green Fluorescent Protein-tagged Cryptococcus neoformans by Immunofluorescence on Paraffin-embedded Brain Sections.
Journal
Applied immunohistochemistry & molecular morphology : AIMM
ISSN: 1533-4058
Titre abrégé: Appl Immunohistochem Mol Morphol
Pays: United States
ID NLM: 100888796
Informations de publication
Date de publication:
01 01 2022
01 01 2022
Historique:
received:
31
03
2021
accepted:
25
08
2021
pubmed:
18
9
2021
medline:
17
3
2022
entrez:
17
9
2021
Statut:
ppublish
Résumé
Cryptococcus neoformans is an important pathogen causing opportunistic fungal meningitis. The pathogenic mechanism of cryptococcal meningitis remains unclear. We aimed to describe a practical approach for studying the pathologic features of cryptococcal central nervous system infection by immunofluorescence on paraffin-embedded brain of mice using different antigen retrieval methods. After 14 days of intratracheal inoculation of green fluorescent protein-tagged C. neoformans (H99-GFP), C57BL/6J mice brains were fixed in 4% paraformaldehyde and embedded in paraffin. Antigen retrieval methods such as microwaves, 1% sodium lauryl sulfate, 1 N HCl, pepsase, and tryptase were used on 5-μm paraffin sections and the effects were compared. The green fluorescence of H99-GFP persisted with antigen retrieval using 1% sodium lauryl sulfate. After immunofluorescent staining, H99-GFP, glial fibrillary acidic protein-tagged astrocytes, and ionized calcium-binding adapter molecule 1-tagged microglia could be observed clearly. Based on our results, we provide a practical approach for the further study of the interaction between C. neoformans and brain cells.
Identifiants
pubmed: 34534990
doi: 10.1097/PAI.0000000000000976
pii: 00129039-202201000-00011
doi:
Substances chimiques
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
72-77Informations de copyright
Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.
Déclaration de conflit d'intérêts
The authors declare no conflict of interest.
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