High-Speed In Vivo Voltage Imaging Dataset of Hippocampal Pyramidal Neurons and Interneurons

Here we present data used to demonstrate Activity Localization Imaging (ALI), a method that visualizes and extracts neuronal voltage signals from densely labeled population. 
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The detailed principles of ALI and the methods for acquiring these data are described in: 
Chen et al., Imaging neuronal voltage beyond the scattering limit, Nature Methods (2025). 
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File descriptions:
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- “<strong>MouseCA1-PyramidalCells.tsm</strong>”: Contains voltage imaging data of CA1 pyramidal neurons (used in Figs. 2-4 of the paper).
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- “<strong>MouseCA1-PV-Interneurons-Targeted-illumination.tsm</strong>”: Contains voltage imaging data of CA1 PV interneurons acquired using targeted illumination (used Fig. 5a, b). This dataset was previously published in Abdelfattah et al. Neuron (2023). 
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Each of these files contains 360,000 images of Voltron 2-labeled neurons (160 x 190 pixels per image) taken at 2000 frames per second. The pixel size is 7.5 x 7.5 micrometers.
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- “<strong>tsm_read.m</strong>”: A MATLAB script for reading these images. 
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- “<strong>MouseCA1-PyramidalCells_2P_zstack.zip</strong>”: Contains a high-resolution two-photon z-stack covering the same pyramidal cells as in “MouseCA1-PyramidalCells.tsm”. 
--- The raw data file (“<strong>2P_verification_rawdata.tif</strong>”) contains both green and red channels. The Voltron fluorescence is visible in the red channel. 
--- To facilitate analysis, the red channel images were extracted, rotated 90 degrees to match the neuron orientation in voltage imaging, and stored in a separate file (“<strong>2P_verification_RedChannel_Rotated.tif</strong>”). 
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The z-stack contains 100 images taken at 2 micrometer steps. 
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