Design and Synthesis of an Activatable Photoacoustic Probe for Hypochlorous Acid.

Photoacoustic (PA) imaging is a novel imaging modality that combines the high contrast of optical imaging and the deep tissue penetration of ultrasound. PA imaging contrast agents targeting various biological phenomena have been reported, but the development of activatable PA probes, which show a PA signal only in the presence of target molecules, remains challenging in spite of their potential usefulness for real-time PA imaging of specific biomolecules in vivo. To establish a simple design strategy for activatable PA probes, we first designed and synthesized a silicon-rhodamine based near-infrared nonfluorescent dye, wsSiNQ660 (water-soluble SiNQ660), as a scaffold and demonstrated that it offers a high conversion efficiency from light to ultrasound compared to typical near-infrared fluorescent dyes. Importantly, absorption off/on strategies previously established for rhodamine-based fluorescent probes are also applicable to this nonfluorescent dye scaffold. We validated this approach by synthesizing an activatable PA probe for hypochlorous acid (HOCl) and confirmed that it enables three-dimensional imaging of HOCl in mouse subcutis.

Quantitative phase imaging by optimized asymmetric illumination.

We have presented a simple approach for quantitative phase imaging by optimizing asymmetric illumination of a conventional microscope. With this illumination, the light intensity modulation accompanying refraction at the surface profile of phase objects occurs, and "phase-gradient information" can be derived by detecting it. Two images with phase-gradient information on different axes are converted into the two-dimensional phase distribution of the specimen by introducing the phase-gradient transfer function, which is the intensity change due to refraction by the phase-gradient of a specimen. We experimentally confirm accurate and repeatable performance of our method and demonstrate phase imaging of live cells.

Method for observing phase objects without halos or directional shadows.

A new microscopy method for observing phase objects without halos or directional shadows is proposed. The method is based on transformation of the surface profile of phase objects into a light intensity pattern. The key optical element is an annular aperture at the front focal plane of a condenser. The light flux passing through the annular aperture is changed by the specimen's surface profile and then passes through an objective and contributes to image formation. Theory and experimental results are presented. Phase images of induced pluripotent stem (iPS) cells with clear outlines were obtained with this method.