μSPIM toolset: a software platform for selective plane illumination microscopy

Saska, Daniel, Pichler, Paul, Qian, Chen, Buckley, Christopher L and Lagnado, Leon (2021) μSPIM toolset: a software platform for selective plane illumination microscopy. Journal of Neuroscience Methods, 347. a108952. ISSN 0165-0270

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Abstract

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, μSPIM Toolset, built around the widely adopted MicroManager platform, that provides control and acquisition functionality for a SPIM. A key advantage of μSPIM 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: μSPIM 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, μSPIM Toolset uses a scanned beam to allow imaging at higher acquisition frequencies while minimizing disturbance of the sample.

Conclusions: The μSPIM 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.

Item Type: Article
Keywords: acquisition, micromanager, selective light-sheet microscopy, spim, toolbox, toolset, uspim, μSPIM, Animals, Lighting, Microscopy, Optical Imaging, Software, Zebrafish
Schools and Departments: School of Engineering and Informatics > Informatics
School of Life Sciences > Neuroscience
SWORD Depositor: Mx Elements Account
Depositing User: Mx Elements Account
Date Deposited: 08 Oct 2021 09:01
Last Modified: 08 Oct 2021 09:15
URI: http://sro.sussex.ac.uk/id/eprint/102205

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