An open-source platform for 3D transcranial Ultrasound Localization Microscopy in awake mice

Abstract We present an open-source code for 3D super-resolution ultrasound imaging. Open-3DULM was applied to transcranial imaging in habituated awake mice. Comparative analysis reveals that isoflurane anesthesia induces significant vasodilation and higher cerebral blood flow compared to the awake state. This method could serve as a reference for developing new types of 3D vascular quantifications, particularly emphasizing the importance of awake and 3D imaging for accurate cerebral blood flow measurement in neuroscience research.