All-optical 3D scanning confocal microscopy incorporating superimposed phase control

The conventional laser confocal microscopy achieves imaging of the three-dimensional volume region by establishing the conjugate relationship between the sample space and the image space, and through the three-dimensional motion of the sample. However, the movement of the sample stage not only reduces the imaging speed and introduces motion artifacts, but also limits the imaging samples to forms such as slices that are compatible with the sample holder, precluding in vivo three-dimensional imaging. In this study, by introducing phase control under end-to-end closed-loop control, precise and aberration-free all-optical three-dimensional high-speed scanning with high numerical aperture excitation is achieved. Without the need for sample movement, two-dimensional imaging at a kHz speed with 300 nm lateral resolution and a three-dimensional volume imaging speed in the hundreds of Hertz are realized, enabling in vivo real-time high-resolution three-dimensional imaging of research objects such as living mice in various life states.