High-speed, multi-modal, label-free imaging of pathological slices with a Bessel beam

Abstract

Optical imaging of stained pathological slices has become the gold standard for disease diagnosis. However, the procedure of sample preparation is complex and time-consuming. Multiphoton microscopy (MPM) is promising for label-free imaging, but the imaging speed is limited, especially for whole slice imaging. Here we propose a high-speed, multi-modal, label-free MPM by Bessel scan-based strip mosaicking. With a Bessel beam for excitation, the extended depth-of-focus not only enables full axial information acquisition at once, but also alleviates the demanding requirement of sample alignment. With the strip mosaicking protocol, we can save the time of frequent sample transferring. Besides, we add a closely-attached reflection mirror under the sample for enhancing epi-detection signals, and employ circularly polarized beams for recording comprehensive information. We demonstrate its application in multi-modal, label-free imaging of human gastric cancer slices and liver cancer slices, and show its potential in rapid disease diagnosis.