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Wavefront shaping techniques are being actively developed to achieve optical focusing through and inside opaque scattering media. These techniques promise to revolutionize biophotonics by enabling deep-tissue non-invasive optical imaging, optogenetics, optical tweezing, and light-based therapy. Among the existing wavefront shaping techniques, optical time-reversal-based techniques determine the optimum wavefront globally based on the principle of time reversal, without the need to perform time-consuming iterations to optimize each mode in sequence. In all previous optical time-reversal-based wavefront shaping experiments, Nyquist sampling criterion was followed so that the scattered light field was well-sampled during wavefront measurement and wavefront reconstruction. In this work, we overturn this conventional practice by demonstrating that a high-quality optical focus can still be achieved even when the scattered light field is under-sampled. Even more strikingly, we show both theoretically and experimentally that the focus achieved by the under-sampling scheme can be one order of magnitude brighter than that achieved by the well-sampling schemes used in previous works, where 3×3 to 5×5 pixels sampled one speckle grain on average. Moreover, since neighboring pixels were uncorrelated in feedback-based wavefront shaping, introducing the concept of sub-Nyquist sampling in time-reversal-based wavefront shaping makes the optimal phase maps obtained using these two different methods consistent. We anticipate that this newly explored under-sampling scheme will transform the understanding of optical time reversal and boost the performance of optical imaging, manipulation, and communication through opaque scattering media.

Focusing light deep inside and through thick biological tissue is critical to many applications. However, optical scattering prevents light from being focused through thick biological tissue, which restricts biophotonics to a limited depth of about 1 mm. To break this optical diffusion limit, digital optical phase conjugation (DOPC) based wavefront shaping techniques are being actively developed. Previous DOPC systems employed spatial light modulators that modulated either the phase or the amplitude of the conjugate light field. Here, we achieve optical focusing through scattering media by using polarization modulation based generalized DOPC. First, we describe an algorithm to extract the polarization map from the measured scattered field. Then, we validate the algorithm through numerical simulations, and find the focusing contrast achieved by polarization modulation is similar to that achieved by phase modulation, and is higher than those achieved by binary-phase and binary-amplitude modulations. Finally, we build a system using an inexpensive twisted nematic liquid crystal based spatial light modulator, and experimentally demonstrate light focusing through 3-mm thick chicken breast tissue. Since the polarization modulation based SLMs are widely used in displays and are having more and more pixel counts with the prevalence of 4K displays, these SLMs are inexpensive and valuable devices for wavefront shaping. Thus, we anticipate that polarization modulation based SLMs will gain their prevalence in the field of wavefront shaping.

Optical phase conjugation (OPC) based wavefront shaping techniques focus light through or within scattering media, which is critically important for deep-tissue optical imaging, manipulation, and therapy. However, to date, the sample thicknesses used in wavefront shaping experiments have been limited to only a few millimeters or several transport mean free paths. Here, by using a long-coherence-length laser and an optimized digital OPC system that efficiently delivers light power, we focused 532 nm light through tissue-mimicking phantoms up to 9.6 cm thick, as well as through ex vivo chicken breast tissue up to 2.5 cm thick.

Optical scattering of biological tissue limits the working depth of conventional biomedical optics, which relies on the detection of ballistic photons. Recent developed optical phase conjugation (OPC) technique breaks through this depth limit by harnessing the scattered photons and shaping an optical wavefront that can “undo” the optical scattering. The OPC system measures the complex light field exiting the tissue and reconstructs a phase conjugated copy of the measured wavefront, which propagates in the reversed direction to the source of the light. To focus light inside a scattering medium, an embedded light source or “guidestar” is often required. Therefore, developing guidestar mechanisms plays an important role in advancing the OPC technique for deep tissue optical focusing and imaging. In addition to having strong optical modulation efficiency and compact size, a favorable guidestar for biomedical applications should also have good biocompatibility, fast response time, and be noninvasive or require only minimally invasive procedure. While a number of guidestar mechanisms have been developed and showed promising for various biomedical applications, they all have their own limitations. We have been developing new guidestars and tailoring them to meet the need for biomedical imaging and therapies. We are going to present our recent progress in novel guidestar development, compare them with established guidestar mechanisms, and discuss their potential in biomedical applications.

Optical scattering of biological tissue limits the penetration depth of conventional optical techniques, which rely on the detection of ballistic photons. Recent developed optical phase conjugation (OPC) technique breaks through this depth limit by shaping an optical wavefront that can “undo” the optical scattering. Assisted with an ultrasound focus, this technique enables optical focusing inside biological tissue in a freely addressable fashion. However, ultrasound modulation efficiency is low and the focusing resolution is limited by the ultrasound. Here we present a new technique, time-reversed ultrasound microbubble encoded (TRUME) optical focusing, which is able to provide high focusing efficiency and sub-ultrasound resolution. This technique achieves the wavefront solution by taking the difference of the optical fields captured outside the sample before and after ultrasound-driven microbubble destruction. A conjugated wavefront was then reconstructed and sent back to the sample to form a focus at the site of microbubble destruction. We experimentally demonstrate that a focus with ~2 um size was formed through a 2-mm thick biological tissue using this method. While the size the microbubble sets the resolution of an individual focus, the scale of the ultrasound focus limits the focusing addressability of this technique. Importantly, by utilizing the nonlinear destruction of microbubbles, the TRUME technique breaks the addressable focus resolution barrier imposed by the ultrasound focus. We experimentally demonstrate a 2-fold improvement in addressability using this effect. Since microbubbles are widely used as ultrasound contrast agents in human, this technique provides a promising solution for focusing light deep inside biological tissue.

Focusing light deep inside scattering media plays a key role in such biomedical applications as high resolution optical imaging, control, and therapy. In recent years, wavefront shaping technologies have come a long way in controlling light propagation in complex media. A prominent example is time-reversed ultrasonically encoded (TRUE) focusing, which allows noninvasive introduction of “guide stars” inside biological tissue to guide light focusing. By measuring the optical wavefront emanating from an ultrasound focus created at the target location, TRUE determines the desired wavefront non-iteratively, and achieves focusing at the target position via a subsequent optical time reversal. Compared to digital counterparts that employ slow electronic spatial light modulators and cameras, analog TRUE focusing relies on nonlinear photorefractive crystals that inherently accommodate more spatial modes and eliminate the troublesome alignment and data transfer required by digital approaches. However, analog TRUE focusing suffers from its small gain, defined as the energy or power ratio between the focusing and probing beams in the focal volume. Here, by implementing a modified analog TRUE focusing scheme that squeezes the duration of the time-reversed photon packet below the carrier-recombination-limited hologram decay time of the crystal, we demonstrated a photon flux amplification much greater than unity at a preset focal voxel in between two scattering layers. Although the energy gain was still below unity, the unprecedented power gain will nevertheless benefit new biomedical applications.

Optical focusing plays a central role in biomedical optical imaging, manipulation, and therapy. However, in scattering media, direct optical focusing becomes infeasible beyond ~10 mean free paths. To break this limit, time-reversed ultrasonically encoded (TRUE) optical focusing phase-conjugates ultrasonically tagged diffuse light back to the ultrasonic focus, thus forming a focus deep inside scattering media. In previous works, the speed of wavefront measurement was limited by the low frame rate of the camera used to record the four images required for phase-shifting holography. Moreover, most of the bits of a pixel value were used to represent an informationless background caused by the large amount of untagged light, increasing the amount of data to transfer and necessitating the use of costly high-resolution analog-to-digital converters (ADCs). Here, we developed a digital TRUE focusing system based on a lock-in camera (300×300 pixels), in which each pixel performs analog lock-in detection on chip. Since only the information of the signal, not that of the background, is digitized, the lock-in camera reduces the amount of data to transfer, and enables the use of cheap low-resolution ADCs. Using this lock-in camera, we were able to measure the wavefront of ultrasonically tagged light in less than 0.3 ms, and to achieve TRUE focusing in between two ground glass diffusers. Even when the signal-to-background ratio dropped to 6.32×10^(-4), a phase sensitivity as low as 0.51 rad could still be realized, which is more than enough for digital optical phase conjugation.

We describe the design and performance of a recently implemented retinal imaging system for the human eye that combines adaptive optics (AO) with spectral domain optical coherence tomography (OCT) and scanning laser ophthalmoscopy (SLO). The AO-OCT-SLO system simultaneously acquires SLO frames and OCT B-scans at 60 Hz with an OCT volume acquisition scan rate of 0.24 Hz. The SLO images are used to correct for eye motion during the registration of OCT B-scans. Key optical design considerations are discussed including: minimizing system aberrations through the use of off-axis relay telescopes; choice of telescope magnification based on pupil plane requirements and restrictions; and the use of dichroic beam splitters to separate and re-combine OCT and SLO beams around the nonshared horizontal scanning mirrors. We include an analysis of closed-loop AO correction on a model eye and compare these findings with system performance in vivo. The 2D and 3D OCT scans included in this work demonstrate the ability of this system to laterally and axially resolve individual cone photoreceptors, while the corresponding SLO images show the en face mosaics at the photoreceptor layer showing rods and cones. Images from both healthy and diseased retina are presented.