Multimodal optical techniques are useful for the comprehensive characterization of material properties. In this work, we developed a new, to the best of our knowledge, multimodal technology that can simultaneously measure a subset of mechanical, optical, and acoustical properties of the sample and is based on the integration of Brillouin (Br) and photoacoustic (PA) microscopy. The proposed technique can acquire co-registered Br and PA signals from the sample. Importantly, using synergistic measurements of the speed of sound and Brillouin shift, the modality offers a new approach to quantifying the optical refractive index, which is a fundamental property of a material and is not accessible by either technique individually. As a proof of concept, we demonstrated the feasibility of integrating the two modalities and acquired the colocalized Br and time-resolved PA signals in a synthetic phantom made out of kerosene and CuSO4 aqueous solution. In addition, we measured the refractive index values of saline solutions and validated the result. Comparison with previously reported data showed a relative error of 0.3%. This further allowed us to directly quantify the longitudinal modulus of the sample with the colocalized Brillouin shift. While the scope of the current work is limited to introducing the combined Br-PA setup for the first time, we envision that this multimodal modality could open a new path for the multi-parametric analysis of material properties.

Multimodal optical imaging techniques are useful for various applications, including imaging biological samples for providing comprehensive material properties. In this work, we developed a new modality that can measure a set of mechanical, optical, and acoustical properties of a sample at microscopic resolution, which is based on the integration of Brillouin (Br) and photoacoustic (PA) microscopy. The proposed multimodal imaging technique not only can acquire co-registered Br and PA signals but also allows us to utilize the sound speed measured by PA to quantify the sample's refractive index, which is a fundamental property of the material and cannot be measured by either technique individually. We demonstrated the colocalization of Br and time-resolved PA signals in a synthetic phantom made of kerosene and CuSO4 aqueous solution. In addition, we measured the refractive index of saline solutions and validated the result against published data with a relative error of 0.3 %. This multimodal Br-PA modality could open a new way for characterizing biological samples in physiological and pathological conditions.

Brillouin microscopy based on spontaneous Brillouin scattering has emerged as a unique elastography technique because of its merit of non-contact, label-free, and high-resolution mechanical imaging of biological cell and tissue. Recently, several new optical modalities based on stimulated Brillouin scattering have been developed for biomechanical research. As the scattering efficiency of the stimulated process is much higher than its counterpart in the spontaneous process, stimulated Brillouin-based methods have the potential to significantly improve the speed and spectral resolution of existing Brillouin microscopy. Here, we review the ongoing technological advancements of three methods, including continuous wave stimulated Brillouin microscopy, impulsive stimulated Brillouin microscopy, and laser-induced picosecond ultrasonics. We describe the physical principle, the representative instrumentation, and biological application of each method. We further discuss the current limitations as well as the challenges for translating these methods into a visible biomedical instrument for biophysics and mechanobiology.

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