Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor.
Journal
The Journal of cell biology
ISSN: 1540-8140
Titre abrégé: J Cell Biol
Pays: United States
ID NLM: 0375356
Informations de publication
Date de publication:
02 09 2019
02 09 2019
Historique:
received:
04
03
2019
revised:
28
06
2019
accepted:
25
07
2019
pubmed:
25
8
2019
medline:
19
5
2020
entrez:
25
8
2019
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31444239
pii: jcb.201903019
doi: 10.1083/jcb.201903019
pmc: PMC6719445
doi:
Substances chimiques
Shelterin Complex
0
TERF2IP protein, human
0
Telomere-Binding Proteins
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
3153-3160Subventions
Organisme : NIEHS NIH HHS
ID : P30 ES010126
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL133668
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM122596
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM127145
Pays : United States
Informations de copyright
© 2019 O'Shaughnessy et al.
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