期刊
JOURNAL OF NEUROSCIENCE METHODS
卷 347, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.jneumeth.2020.108952
关键词
selective light-sheet microscopy; spim; micromanager; acquisition; toolbox; toolset; uspim; mu SPIM
资金
- Wellcome Trust [102905/Z/13/Z]
- Biotechnology and Biological Sciences Research Council [BB/P022197/1]
- BBSRC [BB/P022197/1] Funding Source: UKRI
- Wellcome Trust [102905/Z/13/Z] Funding Source: Wellcome Trust
The mu SPIM Toolset is an open-source software built around the MicroManager platform that optimizes SPIM imaging, allowing high-frequency imaging of neural activity with relative independence from the microscope design. It provides a flexible solution for monitoring neural activity in the larval zebrafish brain.
Background: Selective Plane Illumination Microscopy (SPIM) is a fluorescence imaging technique that allows volumetric imaging at high spatio-temporal resolution to monitor neural activity in live organisms such as larval zebrafish. A major challenge in the construction of a custom SPIM microscope using a scanned laser beam is the control and synchronization of the various hardware components. New Method: We present an open-source software, mu SPIM Toolset, built around the widely adopted MicroManager platform, that provides control and acquisition functionality for a SPIM. A key advantage of mu SPIM Toolset is a series of calibration procedures that optimize acquisition for a given set-up, making it relatively independent of the optical design of the microscope or the hardware used to build it. Results: mu SPIM Toolset allows imaging of calcium activity throughout the brain of larval zebrafish at rates of 100 planes per second with single cell resolution. Comparison with Existing Methods: Several designs of SPIM have been published but are focused on imaging of developmental processes using a slower setup with a moving stage and therefore have limited use for functional imaging. In comparison, mu SPIM Toolset uses a scanned beam to allow imaging at higher acquisition frequencies while minimizing disturbance of the sample. Conclusions: The mu SPIM Toolset provides a flexible solution for the control of SPIM microscopes and demonstrated its utility for brain-wide imaging of neural activity in larval zebrafish.
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