Deformable mirror-based photoacoustic remote sensing (PARS) microscopy for depth scanning.

Optically shifting the focal plane to allow depth scanning of delicate biological structures and processes in their natural environment offers an appealing alternative to conventional mechanical scanning. Our technique uses a deformable mirror-based photoacoustic remote sensing microscopy (PARS) with a focus shifting of Δz ∼ 240 µm. We achieve this by integrating a deformable mirror that functions as a varifocal mirror for axial scanning. First, the system's focal shift capability was demonstrated with USAF resolution targets and carbon fiber phantoms, followed by in-vivo visualizations of blood vessels in chicken embryo chorioallantoic membrane (CAM). This work represents an initial step toward developing a non-contact, label-free, and aberration-free PARS imaging system with axial scanning capability.

Hyperspectral absorption microscopy using photoacoustic remote sensing.

An improved method of remote optical absorption spectroscopy and hyperspectral optical absorption imaging is described which takes advantage of the photoacoustic remote sensing detection architecture. A wide collection of photoacoustic excitation wavelengths ranging from 210 nm to 1550 nm was provided by a nanosecond tunable source allowing access to various salient endogenous chromophores such as DNA, hemeproteins, and lipids. Sensitivity of the device was demonstrated by characterizing the infrared absorption spectrum of water. Meanwhile, the efficacy of the technique was explored by recovering cell nuclei and oxygen saturation from a live chicken embryo model and by recovering adipocytes from freshly resected murine adipose tissue. This represents a continued investigation into the characteristics of the hyperspectral photoacoustic remote sensing technique which may represent an effective means of non-destructive endogenous contrast characterization and visualization.

Functional photoacoustic remote sensing microscopy using a stabilized temperature-regulated stimulated Raman scattering light source.

Stimulated Raman scattering (SRS) has been widely used in functional photoacoustic microscopy to generate multiwavelength light and target multiple chromophores inside tissues. Despite offering a simple, cost-effective technique with a high pulse repetition rate; it suffers from pulse-to-pulse intensity fluctuations and power drift that can affect image quality. Here, we propose a new technique to improve the temporal stability of the pulsed SRS multiwavelength source. We achieve this by lowering the temperature of the SRS medium. The results suggest that a decrease in temperature causes an improvement of temporal stability of the output, considerable rise in the intensity of the SRS peaks, and significant increase of SRS cross section. The application of the method is shown for in vivo functional imaging of capillary networks in a chicken embryo chorioallantois membrane using photoacoustic remote sensing microscopy.

Functional and structural ophthalmic imaging using noncontact multimodal photoacoustic remote sensing microscopy and optical coherence tomography.

Early diagnosis of ocular diseases improves the understanding of pathophysiology and aids in accurate monitoring and effective treatment. Advanced, multimodal ocular imaging platforms play a crucial role in visualization of ocular components and provide clinicians with a valuable tool for evaluating various eye diseases. Here, for the first time we present a non-contact, multiwavelength photoacoustic remote sensing (PARS) microscopy and swept-source optical coherence tomography (SS-OCT) for in-vivo functional and structural imaging of the eye. The system provides complementary imaging contrasts of optical absorption and optical scattering, and is used for simultaneous, non-contact, in-vivo imaging of murine eye. Results of vasculature and structural imaging as well as melanin content in the retinal pigment epithelium layer are presented. Multiwavelength PARS microscopy using Stimulated Raman scattering is applied to enable in-vivo, non-contact oxygen saturation estimation in the ocular tissue. The reported work may be a major step towards clinical translation of ophthalmic technologies and has the potential to advance the diagnosis and treatment of ocular diseases.