216-All-Optical In Situ Histology of Heterogeneous Tissue

The article details the development and application of a robotic nonlinear optical system designed for all-optical in situ histology of heterogeneous tissues. This innovative system combines iterative multiphoton microscopy with spatiotemporal line-focused laser micromachining to achieve submicrometer resolution 3D imaging and reconstruction of large-scale samples, such as the intact murine craniocerebral system. A key advancement is the use of spatiotemporal focusing to increase machining efficiency by 100-fold and the implementation of a smart, autonomous control pipeline for multi-day operation. The research demonstrates the system’s ability to generate high-resolution images of complex structures, like the skull and vasculature, while minimally deforming the tissue compared to traditional mechanical sectioning methods. This technique offers a solution for volumetric imaging of tissues where conventional methods like tissue clearing or vibratome sectioning are limited due to tissue heterogeneity.

References:

• Ji X, Huang S, Friedman B, et al. Spatiotemporal focusing enables all-optical in situ histology of heterogeneous tissue[J]. Nature Methods, 2025: 1-6.