Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae
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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-10-01
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| Series: | BMC Research Notes |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13104-019-4699-9 |
| _version_ | 1818158771924369408 |
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| author | Monika Danchenko Lucia Csaderova Pierre Edouard Fournier Zuzana Sekeyova |
| author_facet | Monika Danchenko Lucia Csaderova Pierre Edouard Fournier Zuzana Sekeyova |
| author_sort | Monika Danchenko |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-11T15:35:23Z |
| format | Article |
| id | doaj.art-524d75d492914bb9a82605e64a4c1b90 |
| institution | Directory Open Access Journal |
| issn | 1756-0500 |
| language | English |
| last_indexed | 2024-12-11T15:35:23Z |
| publishDate | 2019-10-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Research Notes |
| spelling | doaj.art-524d75d492914bb9a82605e64a4c1b902022-12-22T00:59:57ZengBMCBMC Research Notes1756-05002019-10-011211710.1186/s13104-019-4699-9Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiaeMonika Danchenko0Lucia Csaderova1Pierre Edouard Fournier2Zuzana Sekeyova3Biomedical Research Center, Slovak Academy of SciencesBiomedical Research Center, Slovak Academy of SciencesUMR VITROME, AMU, IRD SSA, AP-HM, IHU Mediterranee InfectionBiomedical Research Center, Slovak Academy of SciencesAbstract 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.http://link.springer.com/article/10.1186/s13104-019-4699-9Immunofluorescence assayRickettsiaeCytoskeleton fixationCell cultureActin-based motility |
| spellingShingle | Monika Danchenko Lucia Csaderova Pierre Edouard Fournier Zuzana Sekeyova Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae BMC Research Notes Immunofluorescence assay Rickettsiae Cytoskeleton fixation Cell culture Actin-based motility |
| title | Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae |
| title_full | Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae |
| title_fullStr | Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae |
| title_full_unstemmed | Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae |
| title_short | Optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae |
| title_sort | optimized fixation of actin filaments for improved indirect immunofluorescence staining of rickettsiae |
| topic | Immunofluorescence assay Rickettsiae Cytoskeleton fixation Cell culture Actin-based motility |
| url | http://link.springer.com/article/10.1186/s13104-019-4699-9 |
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