Jian Wei Tay

Ultrasonically encoded wavefront shaping for focusing into random media

Focusing light into opaque random or scattering media such as biological tissue is a much sought-after goal for biomedical applications such as photodynamic therapy, optical manipulation, and photostimulation. However, focusing with conventional lenses is restricted to one transport mean free path in scattering media, limiting both optical penetration depth and resolution. Focusing deeper is possible by using optical phase conjugation or wavefront shaping to compensate for the scattering. For practical applications, wavefront shaping offers the advantage of a robust optical system that is less sensitive to optical misalignment. Here, the phase of the incident light is spatially tailored using a phase-shifting array to pre-compensate for scattering. The challenge, then, is to determine the phase pattern which allows light to be optimally delivered to the target region. Optimization algorithms are typically employed for this purpose, with visible particles used as targets to generate feedback. However, using these particles is invasive, and light delivery is limited to fixed points. Here, we demonstrate a method for non-invasive and dynamic focusing, by using ultrasound encoding as a virtual guide star for feedback to an optimization algorithm. The light intensity at the acoustic focus was increased by an order of magnitude. This technique has broad biomedical applications, such as in optogenetics or photoactivation of drugs.Phase distortions due to scattering in random media restrict optical focusing beyond one transport mean free path. However, scattering can be compensated for by applying a correction to the illumination wavefront using spatial light modulators. One method of obtaining the wavefront correction is by iterative determination using an optimization algorithm. In the past, obtaining a feedback signal required either direct optical access to the target region, or invasive embedding of molecular probes within the random media. Here, we propose using ultrasonically encoded light as feedback to guide the optimization dynamically and non-invasively. In our proof-of-principle demonstration, diffuse light was refocused to the ultrasound focal zone, with a focus-to-background ratio of more than one order of magnitude after 600 iterations. With further improvements, especially in optimization speed, the proposed method should find broad applications in deep tissue optical imaging and therapy.

Amplitude-masked photoacoustic wavefront shaping and application in flowmetry

Optical-resolution photoacoustic flowmetry (PAF) allows noninvasive single-cell flow measurements. However, its operational depth is limited by optical diffusion, which prevents focusing beyond shallow depths in scattering media, as well as reducing the measurement signal-to-noise ratio (SNR). To overcome this limitation, we used binary-amplitude wavefront shaping to enhance light focusing in the presence of scattering. Here, the transmission modes that contributed constructively to the intensity at the optical focus were identified and selectively illuminated, resulting in a 14-fold intensity increase and a corresponding increase in SNR. This technique can potentially extend the operational depth of optical-resolution PAF beyond 1 mm in tissue.

Continuous scanning of a time-reversed ultrasonically encoded optical focus by reflection-mode digital phase conjugation

Time-reversed ultrasonically encoded (TRUE) optical focusing in turbid media was previously implemented using both analog and digital phase conjugation. The digital approach, in addition to its large energy gain, can improve the focal intensity and resolution by iterative focusing. However, performing iterative focusing at each focal position can be time-consuming. Here, we show that by gradually moving the focal position, the TRUE focal intensity is improved, as in iterative focusing at a fixed position, and can be continuously scanned to image fluorescent targets in a shorter time. In addition, our setup is, to the best of our knowledge, the first demonstration of TRUE focusing using a digital phase conjugate mirror in a reflection mode, which is more suitable for practical applications.

Hybridized wavefront shaping for high-speed, high-efficiency focusing through dynamic diffusive media

Abstract. One of the prime limiting factors of optical imaging in biological applications is the diffusion of light by tissue, which prevents focusing at depths greater than the optical diffusion limit (typically ∼1  mm). To overcome this challenge, wavefront shaping techniques that use a spatial light modulator (SLM) to correct the phase of the incident wavefront have recently been developed. These techniques are able to focus light through scattering media beyond the optical diffusion limit. However, the low speeds of typically used liquid crystal SLMs limit the focusing speed. Here, we present a method using a digital micromirror device (DMD) and an electro-optic modulator (EOM) to measure the scattering-induced aberrations, and using a liquid crystal SLM to apply the correction to the illuminating wavefront. By combining phase modulation from an EOM with the DMD’s ability to provide selective illumination, we exploit the DMD’s higher refresh rate for phase measurement. We achieved focusing through scattering media in less than 8 ms, which is sufficiently short for certain in vivo applications, as it is comparable to the speckle correlation time of living tissue.

Optical focusing in scattering media with photoacoustic wavefront shaping (PAWS)

Controllable light delivery to the region of interest is essential to biomedical optical imaging methods like photoacoustic microscopy. It is, however, challenging beyond superficial depths in biological tissue (~1 mm beneath human skin) due to the strong scattering of light that scrambles the photon propagation paths. Recently, optical wavefront shaping has been proposed to modulate the incident light wavefront to compensate for the scattering-induced phase distortions, and consequentially, convey light optimally to a desired location behind or inside turbid media. To reach an optimum wavefront, a searching algorithm is usually required to optimize a feedback signal. In this work, we present our latest explorations, which use photoacoustic signals as the feedback to remotely and non-invasively guide the wavefront shaping process. Our method does not require direct optical access to the target region or the invasive embedding of fluorescence probes inside turbid media. Experimentally, we have demonstrated that diffuse light can be converged to the ultrasound focus by maximizing the amplitude of photoacoustic emissions from the intended absorbing site. Moreover, we show that wavefront-shaped light focusing can enhance existing optical imaging modalities like photoacoustic microscopy, in regard to signal-to-noise ratio, imaging depth, and potentially, resolution.

Digital reflection-mode time-reversed ultrasonically encoded (TRUE) optical focusing

To achieve localized light delivery beyond turbid layers, TRUE optical focusing has been previously implemented by both analog and digital devices. The digital scheme offers a higher energy gain than the analog version. In many biological applications, the reflection-mode configuration, which uses backscattered light from the sample, is more suitable than the transmission-mode configuration. Although reflection-mode analog TRUE focusing has been demonstrated, its digital implementation has not been explored. Here, we report a reflection-mode digital TRUE focusing to concentrate light through a turbid layer. Further, by simply moving the ultrasound focus, we show the systems dynamic focusing capability.

Optical Focusing in Scattering Media with Photoacoustic Wavefront Shaping (PAWS)

Optical focusing beyond superficial depths in biological tissue is challenging due to the strong optical scattering. We show by using photoacoustic signals as feedback for wavefront shaping, diffuse light can be focused in scattering media.

Amplitude-masked photoacoustic wavefront shaping: theory and application in flowmetry

Optical diffusion in scattering media prevents focusing beyond shallow depths, causing optical imaging and sensing to suffer from low optical intensities, resulting in low signal-to-noise ratios (SNR). Here, we demonstrate focusing using a fast binary-amplitude digital micromirror device to characterize the transmission modes of the scattering medium. We then identify and selectively illuminate the transmission modes which contribute constructively to the intensity at the optical focus. Applying this method to photoacoustic flowmetry, we increased the optical intensity at the focus six-fold, and showed that the corresponding increase in SNR allows particle flow to be measured.

Focusing light in scattering media by ultrasonically-encoded wavefront shaping (SEWS)

Wavefront distortion in scattering media can be compensated for using optical wavefront shaping. In this technique, a spatial light modulator (SLM) is used to apply a spatially distributed phase shift to the optical field. A genetic optimization algorithm was used to obtain the SLM pattern which best focuses light within the medium. The target volume is defined by using a focused ultrasound beam to encode light travelling within the acoustic focus. The ultrasonically-encoded light is measured and used as feedback to the algorithm, which then searches for the pattern which maximizes the encoded light intensity. We call this technique ultrasonically-encoded wavefront shaping (SEWS). Using SEWS, we focused light into a scattering medium consisting of ground glass diffuser and a gelatin phantom. The optical intensity at the target was increased by 11 times over the original intensity. These results were validated using fluorescent imaging at the ultrasonic focus.

Focusing Light in Scattering Media by Ultrasonically-Encoded Wavefront Shaping (SEWS)

We focused light into a scattering medium using a spatial light modulator, increasing the optical intensity at the target by 11x. The phase pattern was obtained using a genetic algorithm, with ultrasonically-encoded light as feedback.