Abstract:In order to achieve high-resolution deep-tissue imaging, multiphoton fluorescence microscopy and photoacoustic tomography had been proposed in the past two decades. However, combining the advantages of these two imaging systems to achieve optical-spatial resolution with an ultrasonic-penetration depth is still a field with challenges. In this paper, we investigate the detection of the two-photon photoacoustic ultrasound, and first demonstrate background-free two-photon photoacoustic imaging in a phantom sample. To generate the background-free two-photon photoacoustic signals, we used a high-repetition rate femtosecond laser to induce narrowband excitation. Combining a loss modulation technique, we successfully created a beating on the light intensity, which not only provides pure sinusoidal modulation, but also ensures the spectrum sensitivity and frequency selectivity. By using the lock-in detection, the power dependency experiment validates our methodology to frequencyselect the source of the nonlinearity. This ensures our capability of measuring the background-free two-photon photoacoustic waves by detecting the 2nd order beating signal directly. Furthermore, by mixing the nanoparticles and fluorescence dyes as contrast agents, the two-photon photoacoustic signal was found to be enhanced and detected. In the end, we demonstrate subsurface two-photon photoacoustic bio-imaging based on the optical scanning mechanism inside phantom samples.

Fossil teeth are primary tools in the study of vertebrate evolution, but standard imaging modalities have not been capable of providing high-quality images in dentin, the main component of teeth, owing to small refractive index differences in the fossilized dentin. Our first attempt to use third-harmonic generation (THG) microscopy in fossil teeth has yielded significant submicrometer level anatomy, with an unexpectedly strong signal contrasting fossilized tubules from the surrounding dentin. Comparison between fossilized and extant teeth of crocodilians reveals a consistent evolutionary signature through time, indicating the great significance of THG microscopy in the evolutionary studies of dental anatomy in fossil teeth.

Reciprocity is a fundamental principle of wave physics and directly relates to the symmetry in the transmission through a system when interchanging the input and output. The coherent transmission matrix (TM) is a convenient method to characterize wave transmission through general media. Here, we demonstrate the optical reciprocal nature of complex media by exploring their TM properties. We measured phase-corrected TMs of forward and round-trip propagation in a single polarization state through a looped 1 m-long step-index optical multimode fiber (MMF) to experimentally verify a transpose relationship between the forward and backward transmission. This symmetry impedes straightforward MMF calibration from proximal measurements of the round-trip TM. Furthermore, we show how focusing through the MMF with digital optical phase conjugation is compromised by system loss since time reversibility relies on power conservation. These insights may inform the development of new imaging techniques through complex media and coherent control of waves in photonic systems.