title: Quantification of larval zebrafish motor function in multiwell plates using open-source MATLAB applications.
abstract: This article describes a method to quantify the movements of larval zebrafish in multiwell plates, using the open-source MATLAB applications LSRtrack and LSRanalyze. The protocol comprises four stages: generation of high-quality, flatly illuminated video recordings with exposure settings that facilitate object recognition; analysis of the resulting recordings using tools provided in LSRtrack to optimize tracking accuracy and motion detection; analysis of tracking data using LSRanalyze or custom MATLAB scripts; and implementation of validation controls. The method is reliable, automated and flexible, requires <1 h of hands-on work for completion once optimized and shows excellent signal:noise characteristics. The resulting data can be analyzed to determine the following: positional preference; displacement, velocity and acceleration; and duration and frequency of movement events and rest periods. This approach is widely applicable to the analysis of spontaneous or stimulus-evoked zebrafish larval neurobehavioral phenotypes resulting from a broad array of genetic and environmental manipulations, in a multiwell plate format suitable for high-throughput applications.
authors.affiliation: 1] Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. [2] Department of Neurology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. [3] Tsinghua University Medical School, Beijing, China.
authors.forename: Yangzhong
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: 1] Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. [2] Department of Neurology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Richard T
authors.initials: RT
authors.lastname: Cattley
authors.affiliation: 1] Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. [2] Department of Neurology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Clinton L
authors.initials: CL
authors.lastname: Cario
authors.affiliation: 1] Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. [2] Department of Neurology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
authors.forename: Qing
authors.initials: Q
authors.lastname: Bai
authors.affiliation: 1] Pittsburgh Institute for Neurodegenerative Diseases, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. [2] Department of Neurology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. [3] Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, USA. [4] Geriatric Research, Education and Clinical Center, Pittsburgh Veterans' Affairs Healthcare System, Pittsburgh, Pennsylvania, USA. [5] Department of Neurology, Pittsburgh Veterans' Affairs Healthcare System, Pittsburgh, Pennsylvania, USA.
authors.forename: Edward A
authors.initials: EA
authors.lastname: Burton
mesh_terms: D000818:Animals; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods*; D007814:Larva; D009043:Motor Activity / Q000502:physiology*; D012984:Software; D014741:Video Recording / Q000295:instrumentation / Q000379:methods; D015027:Zebrafish / Q000502:physiology*