In vivo fluorescence microscopy via iterative multi-photon adaptive compensation technique

Area of Science:

• Biomedical Optics
• Neuroimaging
• Adaptive Optics

Background:

• Scattering tissues pose a significant challenge for in vivo optical imaging.
• Previous studies of the iterative multi-photon adaptive compensation technique (IMPACT) focused on fixed tissues.
• Advancements are needed to improve deep tissue imaging and noninvasive brain observation.

Purpose of the Study:

• To demonstrate the advantages of IMPACT for in vivo imaging.
• To report the latest results of IMPACT in biological samples.
• To assess the capability of IMPACT for functional and noninvasive brain imaging.

Main Methods:

• Application of the iterative multi-photon adaptive compensation technique (IMPACT).
• Wavefront measurement and compensation in highly scattering tissues.
• In vivo imaging experiments in awake mice, including functional imaging and imaging through the intact skull.

Main Results:

• IMPACT significantly improves in vivo neuron imaging in the mouse cortex at depths up to ~660 microns.
• Functional imaging of awake mice was successfully achieved using IMPACT.
• Neuron imaging through the intact skull of adult mice was demonstrated, enabling noninvasive optical measurements.

Conclusions:

• IMPACT is a powerful tool for deep in vivo optical imaging in scattering biological tissues.
• The technique facilitates functional neuroimaging in awake animals.
• IMPACT holds promise for noninvasive optical brain imaging in mice.