A Multiregional Functional Ultrasound Dataset of Brain Responses to Innocuous and Noxious Mechanical Stimulation in Awake Mice

This dataset was generated to systematically investigate brain-wide hemodynamic and neuronal responses to graded mechanical stimulation in awake mice using a multimodal neuroimaging approach. Adult male C57BL/6J mice (8–10 weeks old, n = 8) underwent cranial window surgery under sterile conditions, during which the exposed cortex was covered with a polyethylene membrane and an aluminum alloy headplate was implanted to enable stable head fixation. Following a recovery period of at least 7 days, mice were habituated to head fixation prior to imaging to minimize motion artifacts. Functional ultrasound imaging (fUS) was performed using a commercial ultrasound system equipped with a 15 MHz linear array transducer, with power Doppler signals acquired at a temporal resolution of 3.57 Hz and a spatial resolution of 100 μm.Mechanical stimulation was delivered to the plantar surface of the right hind paw using six calibrated von Frey filaments (0.16–4.0 g) in a pseudorandomized design. Each stimulus lasted 10 seconds and was repeated five times per intensity, with inter-trial intervals of at least 50 seconds and 5-minute intervals between intensity blocks to minimize sensitization and order effects. Imaging was conducted across three coronal planes defined according to the Paxinos and Franklin mouse brain atlas (Bregma −0.71 mm, −1.55 mm, and −3.07 mm), corresponding to cortical, thalamic, and midbrain levels. A separate cohort of transgenic mice (n = 8, expressing GCaMP6s) was used for fiber photometry recordings under the same stimulation paradigm, with calcium signals acquired at 40 Hz from the primary somatosensory hindlimb cortex (S1HL) and anterior pretectal nucleus (APT), providing a neuronal reference for the hemodynamic measurements. Data acquisition was conducted over a 6-month period from September 2025 to February 2026.The dataset comprises multiple types of data files. Raw functional ultrasound data are stored in MATLAB (.mat) format and record cerebral blood volume changes across different mice, coronal planes, and stimulus intensities. Processed data include ΔI/I₀ (relative change in power Doppler signal) hemodynamic time-series and ROI-averaged response signals. Functional connectivity data are provided as Pearson correlation matrices derived from ROI signals, representing inter-regional connectivity strength. ROI mask files are registered to the Paxinos and Franklin mouse brain atlas and cover multiple key regions involved in somatosensory and nociceptive processing, including the primary somatosensory hindlimb cortex (S1HL), motor cortex (M), anterior cingulate cortex (ACC), paraventricular thalamus (PVT), ventral posterolateral/ventral posteromedial nuclei (VPL/VPM), anterior pretectal nucleus (APT), periaqueductal gray (PAG), and median raphe nucleus (MRN), among others. In addition, the dataset includes stimulation timing files indicating stimulus onset and offset, atlas registration images (PNG format), and MATLAB scripts for data processing and analysis.Fiber photometry data are provided in Excel (.xlsx) format. Each table contains 1634 rows and 7 columns. The first column represents time (in seconds), with stimulus onset defined as 0 s. Columns 2–7 correspond to the averaged ΔF/F₀ calcium signals under the six mechanical stimulus intensities (in grams, g), representing mean responses across eight mice. The first row serves as column headers, where the first column indicates time and the remaining columns indicate stimulus intensities. This tabular structure captures neuronal activity dynamics across both time and stimulus intensity dimensions.All data are stored in standard formats to ensure accessibility and reproducibility. Functional ultrasound and ROI-related data are provided in MATLAB (.mat) format, which can be accessed using MATLAB (R2020b or later) or Python libraries such as h5py. Fiber photometry data are provided in Excel (.xlsx) format, and atlas images are provided in PNG format. No systematic missing values are present in the dataset. Potential sources of variability include minor motion artifacts during awake imaging and inter-animal differences; these effects were minimized through habituation procedures and standardized experimental protocols. This dataset provides a reliable resource for studies of sensory processing, pain mechanisms, neurovascular coupling, and large-scale brain network dynamics.