Protocol to measure inter-centrosome distance in adherent cells using epifluorescence microscopy
Summary: We present here a protocol to assay the centrosome separation events at late-G2 phase of the cell cycle by immunofluorescence microscopy. We describe the steps required for imaging and measurement of inter-centrosome distance. Here, we use GAS2L1 as an example, but the protocol can be used...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722001071 |
| _version_ | 1818317186512453632 |
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| author | Franco K.C. Au Edna S.W. Lui Robert Z. Qi |
| author_facet | Franco K.C. Au Edna S.W. Lui Robert Z. Qi |
| author_sort | Franco K.C. Au |
| collection | DOAJ |
| description | Summary: We present here a protocol to assay the centrosome separation events at late-G2 phase of the cell cycle by immunofluorescence microscopy. We describe the steps required for imaging and measurement of inter-centrosome distance. Here, we use GAS2L1 as an example, but the protocol can be used to test any protein for a role in centrosome separation and cohesion. The steps below are specific for hTERT RPE-1 cell lines, but other adherent cell lines (e.g., U2OS, MRC-5) are also amenable for this protocol.For complete details on the use and execution of this protocol, please refer to Au et al. (2017) and Au et al. (2020). |
| first_indexed | 2024-12-13T09:33:19Z |
| format | Article |
| id | doaj.art-fc227b3eec264f3bb5b87795b7586266 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-13T09:33:19Z |
| publishDate | 2022-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-fc227b3eec264f3bb5b87795b75862662022-12-21T23:52:27ZengElsevierSTAR Protocols2666-16672022-03-0131101227Protocol to measure inter-centrosome distance in adherent cells using epifluorescence microscopyFranco K.C. Au0Edna S.W. Lui1Robert Z. Qi2Division of Life Science and State Key Laboratory of Molecular Neuroscience, The Hong Kong University of Science and Technology, Hong Kong, ChinaDivision of Life Science and State Key Laboratory of Molecular Neuroscience, The Hong Kong University of Science and Technology, Hong Kong, ChinaDivision of Life Science and State Key Laboratory of Molecular Neuroscience, The Hong Kong University of Science and Technology, Hong Kong, China; Bioscience and Biomedical Engineering Thrust, The Hong Kong University of Science and Technology (Guangzhou), Guangzhou, China; Corresponding authorSummary: We present here a protocol to assay the centrosome separation events at late-G2 phase of the cell cycle by immunofluorescence microscopy. We describe the steps required for imaging and measurement of inter-centrosome distance. Here, we use GAS2L1 as an example, but the protocol can be used to test any protein for a role in centrosome separation and cohesion. The steps below are specific for hTERT RPE-1 cell lines, but other adherent cell lines (e.g., U2OS, MRC-5) are also amenable for this protocol.For complete details on the use and execution of this protocol, please refer to Au et al. (2017) and Au et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166722001071Cell BiologyCell-based AssaysMicroscopy |
| spellingShingle | Franco K.C. Au Edna S.W. Lui Robert Z. Qi Protocol to measure inter-centrosome distance in adherent cells using epifluorescence microscopy STAR Protocols Cell Biology Cell-based Assays Microscopy |
| title | Protocol to measure inter-centrosome distance in adherent cells using epifluorescence microscopy |
| title_full | Protocol to measure inter-centrosome distance in adherent cells using epifluorescence microscopy |
| title_fullStr | Protocol to measure inter-centrosome distance in adherent cells using epifluorescence microscopy |
| title_full_unstemmed | Protocol to measure inter-centrosome distance in adherent cells using epifluorescence microscopy |
| title_short | Protocol to measure inter-centrosome distance in adherent cells using epifluorescence microscopy |
| title_sort | protocol to measure inter centrosome distance in adherent cells using epifluorescence microscopy |
| topic | Cell Biology Cell-based Assays Microscopy |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722001071 |
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