Depth-resolved volumetric two-photon microscopy based on dual Airy beam scanning

Area of Science:

• Biomedical Optics
• Neuroimaging
• Microscopy

Background:

• Traditional two-photon microscopy (TPM) faces limitations in imaging depth and speed.
• Existing volumetric TPM methods, like Bessel-beam-based approaches, have trade-offs in resolution or scanning efficiency.
• Studying neural dynamics requires advanced imaging techniques capable of fast, deep, and high-resolution volumetric acquisition.

Purpose of the Study:

• To demonstrate a novel dual-Airy-beam-scanning-based volumetric two-photon microscopy (TPM) with intrinsic depth-resolving capability.
• To evaluate the performance and advantages of this new TPM technique compared to existing methods.
• To establish a fast and effective imaging tool for observing dynamic biological processes, particularly in neural biology.

Main Methods:

• Utilized a pair of oppositely accelerating Airy beams for sequential sample illumination in a volumetric TPM setup.
• Leveraged the inherent deflection properties of Airy beams to resolve depth information within the sample.
• Quantified the depth-resolving range achievable with the dual-Airy-beam-scanning approach.

Main Results:

• Successfully demonstrated volumetric TPM with depth-resolving capability up to 32 μm.
• Achieved superior depth-resolving capability compared to Bessel-beam-based TPM.
• Showcased reduced scanning times relative to traditional Gaussian-beam-based TPM.

Conclusions:

• The developed depth-resolved volumetric TPM offers significant advantages in both imaging depth and speed.
• This technique presents a promising advancement for fast imaging of dynamic biological processes.
• The dual-Airy-beam-scanning TPM is well-suited for in-situ studies of neural biology dynamics.