Simultaneous multiplane imaging with reverberation multiphoton microscopy

Really nice pre-print by the people at Boston University, leaded by J. Mertz.

Love the idea of generating ~infinite focal spots (until you run out of photons) inside a sample, and using a extremely fast single-pixel detector to recover the signal. Very original way to tackle volumetric imaging in bio-imaging!

Fundamental workflow of the technique. Extracted from Fig. 1 in the manuscript

Simultaneous multiplane imaging with reverberation multiphoton microscopy

by Devin R. Beaulieu, Ian G. Davison, Thomas G. Bifano, and Jerome Mertz, at arXiv.org

Abstract:

Multiphoton microscopy (MPM) has gained enormous popularity over the years for its capacity to provide high resolution images from deep within scattering samples. However, MPM is generally based on single-point laser-focus scanning, which is intrinsically slow. While imaging speeds as fast as video rate have become routine for 2D planar imaging, such speeds have so far been unattainable for 3D volumetric imaging without severely compromising microscope performance. We demonstrate here 3D volumetric (multiplane) imaging at the same speed as 2D planar (single plane) imaging, with minimal compromise in performance. Specifically, multiple planes are acquired by near-instantaneous axial scanning while maintaining 3D micron-scale resolution. Our technique, called reverberation MPM, is well adapted for large-scale imaging in scattering media with low repetition-rate lasers, and can be implemented with conventional MPM as a simple add-on.