Optical imaging has had a central role in elucidating the underlying biological and physiological mechanisms in living specimens owing to its high spatial resolution, molecular specificity and minimal invasiveness. However, its working depth for in vivo imaging is extremely shallow, and thus reactions occurring deep inside living specimens remain out of reach. This problem originates primarily from multiple light scattering caused by the inhomogeneity of tissue obscuring the desired image information. Adaptive optical microscopy, which minimizes the effect of sample-induced aberrations, has to date been the most effective approach to addressing this problem, but its performance has plateaued because it can suppress only lower-order perturbations. To achieve an imaging depth beyond this conventional limit, there is increasing interest in exploiting the physics governing multiple light scattering. New approaches have emerged based on the deterministic measurement and/or control of multiple-scattered waves, rather than their stochastic and statistical treatment. In this Review, we provide an overview of recent developments in this area, with a focus on approaches that achieve a microscopic spatial resolution while remaining useful for in vivo imaging, and discuss their present limitations and future prospects.
Optical microscopy is an indispensable tool in biology and medicine owing to its high spatial resolution, molecular specificity and minimal invasiveness, but it is limited to the interrogation of superficial layers for in vivo imaging.
The intensity of single-scattered waves used in conventional imaging decreases exponentially with depth; thus, the imaging depth limits are set by the detector dynamic range and the efficiency of the gating operations.
Approaches that make deterministic use of the abundant multiple-scattered (MS) waves have been proposed to enable deep optical imaging while maintaining the microscopic spatial resolving power.
Recording and controlling the wavefront of MS waves enables a complex scattering layer to be converted into a focusing lens, leading to the development of an ultrathin endoscope.
Acousto-optic interactions and wavefront sensing and/or control are integrated to exploit the large penetration depth of ultrasound and high spatial resolution of optical imaging.
Reflection-matrix approaches that record and process all MS waves enable the correction of sample-induced aberrations, exploitation of multiple scattering signals and suppression of multiple scattering noise, allowing for imaging at depths greater than those accessible with conventional confocal and adaptive optics microscopy.
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This research was supported by the Institute for Basic Science (grant no. IBS-R023-D1), the National Research Foundation of Korea (grant nos. NRF-2019R1C1C1008175 and NRF-2016R1A6A3A11936389), and the Catholic Medical Center Research Foundation in the programme year of 2018.
The authors declare no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
- Isoplanatic patch
The area over which the wavefront error remains almost the same.
- Guide stars
Bright, point-like light sources or scatterers that provide a wavefront reference for measuring and correcting wavefront distortions in adaptive optics systems.
- Point optimization
A method or algorithm that optimizes the point spread function by minimizing wavefront errors in adaptive optics systems.
- Spatial light modulator
A device that modulates the amplitude, phase or polarization of light waves in space.
- Optical memory effect
The phenomenon that speckle patterns of scattered light through thin and diffusive media are invariant to small tilts or shifts in an incident wavefront of light.
- Digital micromirror devices
Micromirrors used for high-speed, efficient and reliable spatial-light modulation; originally invented to create video displays in digital projectors.
- Epi-detection geometry
An imaging configuration in which an objective is used for both illumination and detection.
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