We demonstrate the application of an extended field of view hybrid microscope, integrating distinct optical and photoacoustic (PA) contrast modes, for the precise three-dimensional anatomy delineation of the ciliary body/iris structures in healthy rabbit eyes ex vivo. The glutaraldehyde-induced autofluorescence and the intrinsic PA signals provided by each of the employed imaging modalities were characterized by a high spatial complementarity, offering thus rich morphological information regarding the pars plana and pars plicata ciliary body portions, the iris, as well as, the attached zonule fiber strands. The bimodal microscopy approach presented could find application on studies involving the ocular accommodation mechanism or pathological ciliary body conditions, as a powerful diagnostic technique contributing to the understanding of ocular physiology and function.
The combined use of a wavefront modulator and a scattering medium forms an "opaque lens" which forces the light to focus tightly. The adaptive focus has the same shape as the correlation function of the original speckle pattern and it can be generated at defined positions with resolution up to hundreds of nanometers. We have demonstrated that manipulating the speckle pattern spatial components can structure the shape of the focus. Exploiting selectively spatial-frequencies from the speckle components we realized opaque lenses able to produce sub-correlation foci and Bessel beams.
By compensating the random phase delay acquired while a light beam crosses a scattering curtain, it is possible to address the light at selected target position beyond the obstacle. An opaque lens can produce foci with a resolution higher than conventional optics if a strongly scattering medium is exploited. In practice, subwavelength resolution is obtained only for weakly transmitting samples. Herein we present a method which allows obtaining tiny bright optical spots even in presence of a minimum amount of scattering (semi-transparent media) in the beam path. Using a High-Pass spatial filter we block the pseudo-ballistic components of the transmitted beam, we are able to gather light on a spot with a diameter which is one third of the typical speckle grain in absence of the filter.
<i>In vivo</i> optical imaging of biological tissue not only requires the development of new theoretical models and experimental procedures, but also the design and construction of realistic tissue-mimicking phantoms. However, most of the phantoms available currently in literature or the market, have either simple geometrical shapes (cubes, slabs, cylinders) or when realistic in shape they use homogeneous approximations of the tissue or animal under investigation. The goal of this study is to develop a non-homogeneous realistic phantom that matches the anatomical geometry and optical characteristics of the mouse head in the visible and near-infrared spectral range. The fabrication of the phantom consisted of three stages. Initially, anatomical information extracted from either mouse head atlases or structural imaging modalities (MRI, XCT) was used to design a digital phantom comprising of the three main layers of the mouse head; the brain, skull and skin. Based on that, initial prototypes were manufactured by using accurate 3D printing, allowing complex objects to be built layer by layer with sub-millimeter resolution. During the second stage the fabrication of individual molds was performed by embedding the prototypes into a rubber-like silicone mixture. In the final stage the detailed phantom was constructed by loading the molds with epoxy resin of controlled optical properties. The optical properties of the resin were regulated by using appropriate quantities of India ink and intralipid. The final phantom consisted of 3 layers, each one with different absorption and scattering coefficient (μ<sub>a</sub>,μ<sub>s</sub>) to simulate the region of the mouse brain, skull and skin.