Integrated two-photon and optoacoustic microscopy for functional neuroimaging

Area of Science:

• Neuroscience
• Biomedical Engineering
• Medical Imaging

Background:

• Cerebral vasculature-neuron interactions are vital for brain function and neurological disorders.
• Current neuroimaging methods have limitations in resolution, depth, and spatiotemporal alignment for in vivo studies.
• A comprehensive understanding of neurovascular coupling requires advanced imaging techniques.

Purpose of the Study:

• To develop and validate a dual-modality imaging system combining optical-resolution optoacoustic microscopy and two-photon fluorescence microscopy.
• To overcome the limitations of existing functional neuroimaging approaches for in vivo studies.
• To enable simultaneous, high-resolution imaging of both microvasculature and neurons in the mouse cortex.

Main Methods:

• Developed a novel dual-modality system integrating optical-resolution optoacoustic microscopy (OR-PAM) and two-photon fluorescence microscopy (2PFM).
• Employed a semi-simultaneous acquisition protocol for alternating data capture across time and depth planes.
• Utilized the system for in vivo imaging of microcapillaries and neurons in the mouse cortex.

Main Results:

• Achieved submicron resolution imaging of microcapillaries up to 140 μm depth.
• Enabled imaging of neurons beyond 300 μm depth in the mouse cortex.
• Ensured spatiotemporal alignment and robust co-registration of multimodal datasets, minimizing motion artifacts.

Conclusions:

• The developed dual-modality system provides complementary information for comprehensive neurovascular studies.
• This advanced imaging approach enhances the study of neurovascular coupling in both healthy and diseased states.
• The system offers improved capabilities for in vivo investigation of brain function and neurological disorders.