High-throughput volumetric adaptive optical imaging using compressed time-reversal matrix

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 x 40 microns (176 x 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 x 128 x 125 microns (568 x 568 x 125 voxels) in 3.58 s, correcting tissue aberrations at each and every 1-micron depth section, and visualizing myelinated axons with a lateral resolution of 0.45 microns and an axial resolution of 2 microns.

Laser scanning reflection-matrix microscopy for label-free in vivo imaging of a mouse brain through an intact skull

We present a laser scanning reflection-matrix microscopy combining the scanning of laser focus and the wide-field mapping of the electric field of the backscattered waves for eliminating higher-order aberrations even in the presence of strong multiple light scattering noise. Unlike conventional confocal laser scanning microscopy, we record the amplitude and phase maps of reflected waves from the sample not only at the confocal pinhole, but also at other non-confocal points. These additional measurements lead us to constructing a time-resolved reflection matrix, with which the sample-induced aberrations for the illumination and detection pathways are separately identified and corrected. We realized in vivo reflectance imaging of myelinated axons through an intact skull of a living mouse with the spatial resolution close to the ideal diffraction limit. Furthermore, we demonstrated near-diffraction-limited multiphoton imaging through an intact skull by physically correcting the aberrations identified from the reflection matrix. The proposed method is expected to extend the range of applications, where the knowledge of the detailed microscopic information deep within biological tissues is critical.

Fluorescence spectra of atoms in a phase-stabilized magneto-optical trap as an optical lattice

We present a study on characteristics of a magneto-optical trap (MOT) as an optical lattice. Fluorescence spectra of atoms trapped in a MOT with a passively phase-stabilized beam configuration have been measured by means of the photon-counting heterodyne spectroscopy. We observe a narrow Rayleigh peak and well-resolved Raman sidebands in the fluorescence spectra which clearly show that the MOT itself behaves as a three-dimensional optical lattice. Optical-lattice-like properties of the phase-stabilized MOT such as vibrational frequencies and lineshapes of Rayleigh peak and Raman sidebands are investigated systematically for various trap conditions.