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Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor
Lattice light-sheet microscopy (LLSM) is valuable for its combination of reduced photobleaching and outstanding spatiotemporal resolution in 3D. Using LLSM to image biosensors in living cells could provide unprecedented visualization of rapid, localized changes in protein conformation or posttransla...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Rockefeller University Press
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719445/ https://www.ncbi.nlm.nih.gov/pubmed/31444239 http://dx.doi.org/10.1083/jcb.201903019 |
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author | O’Shaughnessy, Ellen C. Stone, Orrin J. LaFosse, Paul K. Azoitei, Mihai L. Tsygankov, Denis Heddleston, John M. Legant, Wesley R. Wittchen, Erika S. Burridge, Keith Elston, Timothy C. Betzig, Eric Chew, Teng-Leong Adalsteinsson, David Hahn, Klaus M. |
author_facet | O’Shaughnessy, Ellen C. Stone, Orrin J. LaFosse, Paul K. Azoitei, Mihai L. Tsygankov, Denis Heddleston, John M. Legant, Wesley R. Wittchen, Erika S. Burridge, Keith Elston, Timothy C. Betzig, Eric Chew, Teng-Leong Adalsteinsson, David Hahn, Klaus M. |
author_sort | O’Shaughnessy, Ellen C. |
collection | PubMed |
description | Lattice light-sheet microscopy (LLSM) is valuable for its combination of reduced photobleaching and outstanding spatiotemporal resolution in 3D. Using LLSM to image biosensors in living cells could provide unprecedented visualization of rapid, localized changes in protein conformation or posttranslational modification. However, computational manipulations required for biosensor imaging with LLSM are challenging for many software packages. The calculations require processing large amounts of data even for simple changes such as reorientation of cell renderings or testing the effects of user-selectable settings, and lattice imaging poses unique challenges in thresholding and ratio imaging. We describe here a new software package, named ImageTank, that is specifically designed for practical imaging of biosensors using LLSM. To demonstrate its capabilities, we use a new biosensor to study the rapid 3D dynamics of the small GTPase Rap1 in vesicles and cell protrusions. |
format | Online Article Text |
id | pubmed-6719445 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-67194452020-03-02 Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor O’Shaughnessy, Ellen C. Stone, Orrin J. LaFosse, Paul K. Azoitei, Mihai L. Tsygankov, Denis Heddleston, John M. Legant, Wesley R. Wittchen, Erika S. Burridge, Keith Elston, Timothy C. Betzig, Eric Chew, Teng-Leong Adalsteinsson, David Hahn, Klaus M. J Cell Biol Research Articles Lattice light-sheet microscopy (LLSM) is valuable for its combination of reduced photobleaching and outstanding spatiotemporal resolution in 3D. Using LLSM to image biosensors in living cells could provide unprecedented visualization of rapid, localized changes in protein conformation or posttranslational modification. However, computational manipulations required for biosensor imaging with LLSM are challenging for many software packages. The calculations require processing large amounts of data even for simple changes such as reorientation of cell renderings or testing the effects of user-selectable settings, and lattice imaging poses unique challenges in thresholding and ratio imaging. We describe here a new software package, named ImageTank, that is specifically designed for practical imaging of biosensors using LLSM. To demonstrate its capabilities, we use a new biosensor to study the rapid 3D dynamics of the small GTPase Rap1 in vesicles and cell protrusions. Rockefeller University Press 2019-09-02 2019-08-23 /pmc/articles/PMC6719445/ /pubmed/31444239 http://dx.doi.org/10.1083/jcb.201903019 Text en © 2019 O'Shaughnessy et al. http://www.rupress.org/terms/https://creativecommons.org/licenses/by-nc-sa/4.0/This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Research Articles O’Shaughnessy, Ellen C. Stone, Orrin J. LaFosse, Paul K. Azoitei, Mihai L. Tsygankov, Denis Heddleston, John M. Legant, Wesley R. Wittchen, Erika S. Burridge, Keith Elston, Timothy C. Betzig, Eric Chew, Teng-Leong Adalsteinsson, David Hahn, Klaus M. Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor |
title | Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor |
title_full | Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor |
title_fullStr | Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor |
title_full_unstemmed | Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor |
title_short | Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor |
title_sort | software for lattice light-sheet imaging of fret biosensors, illustrated with a new rap1 biosensor |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719445/ https://www.ncbi.nlm.nih.gov/pubmed/31444239 http://dx.doi.org/10.1083/jcb.201903019 |
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