Voltage imaging and optogenetics reveal behaviour-dependent changes in hippocampal dynamics. Ultrafast two-photon imaging of a high-gain voltage indicator in awake behaving mice. Bright and photostable chemigenetic indicators for extended in vivo voltage imaging. Fast and sensitive GCaMP calcium indicators for imaging neural populations. High-speed, cortex-wide volumetric recording of neuroactivity at cellular resolution using light beads microscopy. Real-time volumetric microscopy of in vivo dynamics and large-scale samples with SCAPE 2.0. A large field of view two-photon mesoscope with subcellular resolution for in vivo imaging. Nature Reviews Methods Primers 2, 67 (2022). Two-photon calcium imaging of neuronal activity. Grienberger, C., Giovannucci, A., Zeiger, W. Finally, we present an online experimental scenario to provide guidance in setting FIOLA parameters and to highlight the trade-offs of our approach.
We demonstrate reliable and scalable performance of FIOLA on both simulated and real calcium and voltage imaging datasets. FIOLA exploits algorithms optimized for parallel processing on GPUs and CPUs. We therefore developed FIOLA, a framework for fluorescence imaging online analysis that extracts neuronal activity from calcium and voltage imaging movies at speeds one order of magnitude faster than state-of-the-art methods. Reliable extraction of neural activity from fluorescence imaging frames at speeds compatible with indicator dynamics and imaging modalities poses a challenge. This gap not only prevents the execution of real-time closed-loop experiments, but also hampers fast experiment–analysis–theory turnover for high-throughput imaging modalities. However, the lack of corresponding advances in online algorithms has slowed progress in retrieving information about neural activity during or shortly after an experiment. Optical microscopy methods such as calcium and voltage imaging enable fast activity readout of large neuronal populations using light.
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