Abstract
Deep-tissue optical imaging suffers from the reduction of resolving power due to tissue-induced optical aberrations and multiple scattering noise. Reflection matrix approaches recording the maps of backscattered waves for all the possible orthogonal input channels have provided formidable solutions for removing severe aberrations and recovering the ideal diffraction-limited spatial resolution without relying on fluorescence labeling and guide stars. However, measuring the full input–output response of the tissue specimen is time-consuming, making the real-time image acquisition difficult. Here, we present the use of a time-reversal matrix, instead of the reflection matrix, for fast high-resolution volumetric imaging of a mouse brain. The time-reversal matrix reduces two-way problem to one-way problem, which effectively relieves the requirement for the coverage of input channels. Using a newly developed aberration correction algorithm designed for the time-reversal matrix, we demonstrated the correction of complex aberrations using as small as 2% of the complete basis while maintaining the image reconstruction fidelity comparable to the fully sampled reflection matrix. Due to nearly 100-fold reduction in the matrix recording time, we could achieve real-time aberration-correction imaging for a field of view of 40 × 40 µm2 (176 × 176 pixels) at a frame rate of 80 Hz. Furthermore, we demonstrated high-throughput volumetric adaptive optical imaging of a mouse brain by recording a volume of 128 × 128 × 125 µm3 (568 × 568 × 125 voxels) in 3.58 s, correcting tissue aberrations at each and every 1 µm depth section, and visualizing myelinated axons with a lateral resolution of 0.45 µm and an axial resolution of 2 µm.
Highlights
An arbitrary optical system interacting with light waves can be described by transmission and reflection matrices, as far as the linear light-matter interaction is concerned
The reflection matrix containing full optical input–output response of a scattering medium has offered robust image reconstruction in comparison with conventional adaptive optics approaches relying on partial information
It enables the correction of extremely complex aberrations in stringent conditions where there are strong multiple scattering noise and no guide stars available
Summary
An arbitrary optical system interacting with light waves can be described by transmission and reflection matrices, as far as the linear light-matter interaction is concerned. The reflection matrix approaches provide formidable solutions in the context of computational adaptive optics (AO) microscopy[15,16] in that they can deal with extremely severe aberrations with no need for fluorescence labeling and guide stars. This space-domain reflection matrix study proved that it can serve as a type of wavefront sensorless AO17,18 that is combined with hardware correction of aberration by wavefront shaping devices such as a spatial light modulator and deformable mirror to realize ideal diffraction-limited multi-photon fluorescence imaging through an intact skull[14]
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