Multimodal Imaging Breakthrough Enables Whole-Cortex Neural and Vascular Monitoring in Awake Mice

Revolutionary Microscopy Platform Overcomes Longstanding Neuroimaging Challenges

Researchers have developed an innovative multimodal microscope that simultaneously captures neural activity and hemodynamic responses across the entire mouse cortex with unprecedented speed and resolution. The technology, termed multiScope, represents a significant advancement in neurovascular imaging by integrating three complementary modalities: widefield calcium fluorescence microscopy, optical-resolution photoacoustic microscopy (OR-PAM), and laser speckle contrast imaging (LSCI). This breakthrough enables researchers to study brain-wide neural-vascular interactions in awake, behaving animals with single-vessel resolution.

Addressing Fundamental Limitations in Rotary Scanning Systems

The core innovation lies in a novel uniform rotary scanning mechanism that eliminates inherent oversampling problems through ultrafast laser modulation. Traditional polar-coordinate scanning systems suffer from nonuniform sampling that reduces pulse efficiency and causes central region overheating. The research team tackled this through a compound strategy combining ultrafast laser modulation, adaptive vascular excitation, and deep-learning-based sparse sampling reconstruction.

The laser modulation system dynamically disables pulse output at centrally oversampled pixels, reducing the normalized oversampling factor from 1 to 0 and decreasing average power density in the center area by 34%. This innovation prevents thermal damage during long-term imaging sessions, even when using higher pulse energies that would typically cause tissue damage.

Advanced Signal Processing and Adaptive Excitation

The system incorporates sophisticated computational methods to enhance imaging performance. An adaptive photoacoustic excitation scheme uses feature-based alignment models and U-Net segmentation to identify blood vessel patterns and generate laser modulation sequences that disable pulses in non-vascular areas. This approach minimizes unnecessary energy deposition while maintaining image quality., according to additional coverage

Further enhancement comes from a sparse sampling strategy powered by transformer-based deep learning algorithms. The system recovers high-quality images from sparsely sampled raw data, producing smoother vessel boundaries, fewer artifacts, and higher signal-to-noise ratios while significantly reducing sampling points and preventing heat damage.

Multimodal Integration and Technical Specifications

The multiScope platform achieves remarkable technical performance through careful optical engineering:, according to related news

• Cortex-wide field of view: Ø 8.6 mm, covering the entire mouse cortex
• High spatial resolution: 10.7 ± 3.1 μm for fluorescence imaging, 7.1 ± 0.8 μm for OR-PAM
• Ultrafast imaging speed: Up to 4 Hz for OR-PAM, 16.6 Hz for widefield modalities
• Compact footprint: 60 cm × 80 cm × 110 cm, comparable to conventional upright microscopes

The infinity-corrected rotary scan engine makes OR-PAM compatible with widefield imaging by employing separate scan engines that minimize water damping forces during acoustic scanning. This design enables simultaneous operation of all three imaging modalities without compromising performance., according to recent studies

In Vivo Validation and Research Applications

Researchers validated the system using transgenic mice expressing GCaMP6f calcium indicators in cortical neurons. The multiScope successfully captured whole-cortex neural activity, blood vessel architecture, and cerebral blood flow velocity in awake mice during various behaviors including rest, running, and grooming., according to expert analysis

In experimental demonstrations, the system monitored neurovascular coupling during anesthesia induction and electric shock-induced epilepsy, revealing dynamic relationships between neural activity and hemodynamic responses. The platform’s capability for long-term imaging sessions exceeding 30 minutes without tissue damage opens new possibilities for studying neurological conditions and drug effects.

Implications for Neuroscience and Medical Research

This technological advancement addresses critical limitations in current neuroimaging approaches by providing simultaneous, high-resolution monitoring of both neural and vascular activity across large brain areas. The ability to study awake, behaving animals eliminates confounding effects of anesthesia and enables researchers to investigate brain function under more natural conditions., as previous analysis

The multiScope platform represents a significant step forward in understanding neurovascular coupling mechanisms, with potential applications in stroke research, epilepsy studies, neurodegenerative disease investigation, and pharmaceutical development. By providing comprehensive, simultaneous monitoring of multiple physiological parameters, this technology may accelerate discoveries in fundamental neuroscience and therapeutic development.

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