Multiphoton calcium imaging is one of the most powerful tools in modern neuroscience. However, multiphoton data require significant pre-processing of images and post-processing of extracted signals. As a result, many algorithms and pipelines have been developed for the analysis of multiphoton data, particularly two-photon imaging data. Most current studies use one of several algorithms and pipelines that are published and publicly available, and add customized upstream and downstream analysis elements to fit the needs of individual researchers. The vast differences in algorithm choices, parameter settings, pipeline composition, and data sources combine to make collaboration difficult, and raise questions about the reproducibility and robustness of experimental results. We present our solution, called NeuroWRAP (www.neurowrap.org), which is a tool that wraps multiple published algorithms together, and enables integration of custom algorithms. It enables development of collaborative, shareable custom workflows and reproducible data analysis for multiphoton calcium imaging data enabling easy collaboration between researchers. NeuroWRAP implements an approach to evaluate the sensitivity and robustness of the configured pipelines. When this sensitivity analysis is applied to a crucial step of image analysis, cell segmentation, we find a substantial difference between two popular workflows, CaImAn and Suite2p. NeuroWRAP harnesses this difference by introducing consensus analysis, utilizing two workflows in conjunction to significantly increase the trustworthiness and robustness of cell segmentation results.
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http://dx.doi.org/10.3389/fninf.2023.1082111 | DOI Listing |
bioRxiv
October 2024
Department of Biological Sciences, University of Illinois Chicago, 60607.
Sensory systems must perform the dual and opposing tasks of being sensitive to weak stimuli while also maintaining information content in dense and variable sensory landscapes. This occurs in the olfactory system, where OSNs are highly sensitive to low concentrations of odors and maintain discriminability in complex odor environments. How olfactory sensory neurons (OSNs) maintain both sensitivity and sparsity is not well understood.
View Article and Find Full Text PDFMethods Mol Biol
October 2024
Institute of Anatomy, University of Zurich, Zürich, Switzerland.
Methods Mol Biol
October 2024
Department of Neurobiology, University of Pittsburgh, Pittsburgh, PA, USA.
Two-photon microscopy enables imaging of calcium signaling at cellular or subcellular resolution up to hundreds of microns deep in the living brain. Changes in the brightness of fluorescent calcium indicators provide a readout of calcium levels over time, affording information about neuronal activity and/or calcium-dependent subcellular signaling. Here, we describe a protocol for repeated two-photon imaging of calcium signals in mice expressing a genetically encoded calcium indicator that have been implanted with a chronic cranial window.
View Article and Find Full Text PDFSTAR Protoc
December 2024
Department of Psychiatry, University of North Carolina at Chapel Hill, Chapel Hill, NC 27514, USA; Neuroscience Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27514, USA; Department of Applied Physical Sciences, University of North Carolina at Chapel Hill, Chapel Hill, NC 27514, USA; Carolina Stress Initiative, University of North Carolina at Chapel Hill, Chapel Hill, NC 27514, USA; Carolina Institute for Development Disorders, University of North Carolina at Chapel Hill, Chapel Hill, NC 27514, USA; Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27514, USA. Electronic address:
A wide selection of behavioral assays in systems neuroscience relies on head-fixation protocols to integrate in vivo multi-photon imaging approaches. For this, simultaneous pupillometry and locomotion tracking in head-fixed mice are used to measure behavioral responses and identify neural correlates. Here, we present an open-source protocol for assembling a complete head-fixation system that integrates pupillometry and locomotion-estimated tracking with multi-photon calcium imaging.
View Article and Find Full Text PDFPharmacol Rep
December 2024
Cancer Neurophysiology Group, Łukasiewicz - PORT, Polish Center for Technology Development, Stabłowicka 147, Wrocław, 54-066, Poland.
In recent years, fluorescent sensors are enjoying a surge of popularity in the field of neuroscience. Through the development of novel genetically encoded sensors as well as improved methods of detection and analysis, fluorescent sensing has risen as a new major technique in neuroscience alongside molecular, electrophysiological, and imaging methods, opening up new avenues for research. Combined with multiphoton microscopy and fiber photometry, these sensors offer unique advantages in terms of cellular specificity, access to multiple targets - from calcium dynamics to neurotransmitter release to intracellular processes - as well as high capability for in vivo interrogation of neurobiological mechanisms underpinning behavior.
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