In vivo volumetric imaging of biological dynamics in deep tissue via wavefront engineering.

Lingjie Kong,Meng Cui,Jianyong Tang

doi:10.1364/oe.24.001214

Abstract

Biological systems undergo dynamical changes continuously which span multiple spatial and temporal scales. To study these complex biological dynamics in vivo, high-speed volumetric imaging that can work at large imaging depth is highly desired. However, deep tissue imaging suffers from wavefront distortion, resulting in reduced Strehl ratio and image quality. Here we combine the two wavefront engineering methods developed in our lab, namely the optical phase-locked ultrasound lens based volumetric imaging and the iterative multiphoton adaptive compensation technique, and demonstrate in vivo volumetric imaging of microglial and mitochondrial dynamics at large depth in mouse brain cortex and lymph node, respectively.

Highlights

The hallmark of life is its dynamics [1, 2]
As many of the important applications of optical microscopy such as the calcium imaging demands high signal-to-noise ratio (SNR) [8], spatially parallel detection methods are fundamentally limited in imaging depth
After going through the polarization beam splitter (PBS), quarter wave plate (QWP) and ultrasound lens (UL, TAG lens 2.0, TAG Optics), the beams were imaged onto the end mirror (M1) by a pair of relay lens (RL1 and RL1’)

Summary

Introduction

The hallmark of life is its dynamics [1, 2]. To study the orchestrated cellular signaling and intercellular interplays in vivo, one needs 3D imaging methods that can sample near the natural spatiotemporal resolutions. The parallel detection scheme of these methods benefits high-speed imaging. It only works well for transparent biological systems [7], such as cultured cells, C Elegans and zebrafish larvae. In turbid tissue, such as mouse brain and lymph node, the optical aberrations and scattering induce cross-talk in wide-field recording methods and limit the imaging depth. As many of the important applications of optical microscopy such as the calcium imaging demands high SNR [8], spatially parallel detection methods are fundamentally limited in imaging depth

Methods

Results

Conclusion