Utilize este identificador para referenciar este registo:
https://hdl.handle.net/1822/43632
Título: | Inducible fluorescent speckle microscopy |
Autor(es): | Pereira, António J. Aguiar, Paulo Belsley, M. Maiato, Hélder |
Palavras-chave: | Animals Cell Line, Transformed Drosophila Humans Imaging, Three-Dimensional Metaphase Microscopy, Fluorescence Microtubules Spindle Apparatus |
Data: | 18-Jan-2016 |
Editora: | Rockefeller University Press |
Revista: | Journal of Cell Biology |
Citação: | Pereira, A. J., Aguiar, P., Belsley, M., & Maiato, H. (2016). Inducible fluorescent speckle microscopy. Journal of Cell Biology, 212(2), 245-255. doi: 10.1083/jcb.201506128 |
Resumo(s): | The understanding of cytoskeleton dynamics has benefited from the capacity to generate fluorescent fiducial marks on cytoskeleton components. Here we show that light-induced imprinting of three-dimensional (3D) fluorescent speckles significantly improves speckle signal and contrast relative to classic (random) fluorescent speckle microscopy. We predict theoretically that speckle imprinting using photobleaching is optimal when the laser energy and fluorophore responsivity are related by the golden ratio. This relation, which we confirm experimentally, translates into a 40% remaining signal after speckle imprinting and provides a rule of thumb in selecting the laser power required to optimally prepare the sample for imaging. This inducible speckle imaging (ISI) technique allows 3D speckle microscopy to be performed in readily available libraries of cell lines or primary tissues expressing fluorescent proteins and does not preclude conventional imaging before speckle imaging. As a proof of concept, we use ISI to measure metaphase spindle microtubule poleward flux in primary cells and explore a scaling relation connecting microtubule flux to metaphase duration. |
Tipo: | Artigo |
URI: | https://hdl.handle.net/1822/43632 |
DOI: | 10.1083/jcb.201506128 |
ISSN: | 0021-9525 |
Arbitragem científica: | yes |
Acesso: | Acesso aberto |
Aparece nas coleções: | CDF - FAMO - Artigos/Papers (with refereeing) |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
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J Cell Biol-2016-Pereira-245-55.pdf | 1,81 MB | Adobe PDF | Ver/Abrir |