Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae

Autor: Monika Danchenko, Lucia Csaderova, Pierre Edouard Fournier, Zuzana Sekeyova
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Zdroj: BMC Research Notes, Vol 12, Iss 1, Pp 1-7 (2019)
Druh dokumentu: article
ISSN: 1756-0500
DOI: 10.1186/s13104-019-4699-9
Popis: 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.
Databáze: Directory of Open Access Journals
Nepřihlášeným uživatelům se plný text nezobrazuje