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用于 FRET 生物传感器晶格光片成像的软件，用一个新的 Rap1 生物传感器进行说明。

Software for lattice light-sheet imaging of FRET biosensors, illustrated with a new Rap1 biosensor.

机构信息

Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC.

Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA.

出版信息

J Cell Biol. 2019 Sep 2;218(9):3153-3160. doi: 10.1083/jcb.201903019. Epub 2019 Aug 23.

DOI:10.1083/jcb.201903019
PMID:31444239
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6719445/
Abstract

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.

摘要

晶格层光片显微镜（LLSM）在减少光漂白和在 3D 中具有出色的时空分辨率方面具有价值。在活细胞中使用 LLSM 对生物传感器进行成像，可以提供对蛋白质构象或翻译后修饰快速、局部变化的前所未有的可视化。然而，对于许多软件包来说，使用 LLSM 对生物传感器进行成像所需的计算操作具有挑战性。即使对于重新定向细胞渲染或测试用户可选设置的效果等简单变化，计算也需要处理大量数据，而晶格成像在阈值处理和比率成像方面存在独特的挑战。我们在这里描述了一个名为 ImageTank 的新软件包，它专门用于使用 LLSM 对生物传感器进行实际成像。为了展示其功能，我们使用一种新的生物传感器来研究小 GTPase Rap1 在小泡和细胞突起中的快速 3D 动力学。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d41/6719445/f2171edb757e/JCB_201903019_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d41/6719445/45bece564618/JCB_201903019_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d41/6719445/c185ae786a2a/JCB_201903019_Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d41/6719445/bfa847ff0dbb/JCB_201903019_Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d41/6719445/f2171edb757e/JCB_201903019_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d41/6719445/45bece564618/JCB_201903019_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d41/6719445/c185ae786a2a/JCB_201903019_Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d41/6719445/bfa847ff0dbb/JCB_201903019_Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d41/6719445/f2171edb757e/JCB_201903019_Fig4.jpg

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