Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae.
BMC Res Notes
; 12(1): 657, 2019 Oct 16.
Article
em En
| MEDLINE
| ID: mdl-31619275
ABSTRACT
OBJECTIVE:
The objective was to investigate fixative solutions 3.7% formaldehyde, 4% paraformaldehyde, 4% paraformaldehyde in the cytoskeletal buffer and 4% paraformaldehyde in PHEM buffer (containing PIPES, HEPES, EGTA and MgCl2), applicable for immunofluorescence assay.RESULTS:
Herein we optimized this serological technique, testing four fixative solutions, for the sensitive detection of rickettsial antigens, and preservation of intracellular structures of the host cells, particularly filamentous actin. Rickettsial antigens were presented equally well both with formaldehyde and all paraformaldehyde-based fixations, but only protocol with 4% paraformaldehyde in PHEM buffer allowed accurate imaging of actin filaments, and simultaneously allows monitoring of rickettsiae using actin-based motility during infection inside the host cells.Palavras-chave
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Base de dados:
MEDLINE
Assunto principal:
Rickettsia
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Infecções por Rickettsia
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Citoesqueleto de Actina
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Técnica Indireta de Fluorescência para Anticorpo
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article