Abstract:We demonstrate a high-speed method to image objects through thin scattering media and around corners. The method employs a reference object of known shape to retrieve the speckle-like point spread function of the scatterer. We extract the point spread function of the scatterer from a dynamic scene that includes a static reference object and uses this to image the dynamic objects. Sharp images are reconstructed from the transmission through a diffuser and from the reflection off a rough surface. The sharp and clean reconstructed images from single shot data exemplify the robustness of the method.
Wavefront shaping is increasingly being used in modern microscopy to obtain high-resolution images deep inside inhomogeneous media. Wavefront shaping methods typically rely on the presence of a “guide star” to find the optimal wavefront to mitigate the scattering of light. However, the use of guide stars poses severe limitations. Notably, only objects in the close vicinity of the guide star can be imaged. Here, we introduce a guide-star-free wavefront shaping method in which the optimal wavefront is computed using a digital model of the sample. The refractive index model of the sample, that serves as the input for the computation, is constructed in situ by the microscope itself. In a proof of principle imaging experiment, we demonstrate a large improvement in the two-photon fluorescence signal through a diffuse medium, outperforming state-of-the-art wavefront shaping by a factor of two in imaging depth.
Incoherently illuminated or luminescent objects give rise to a low-contrast speckle-like pattern when observed through a thin diffusive medium, as such a medium effectively convolves their shape with a speckle-like point spread function (PSF). This point spread function can be extracted in the presence of a reference object of known shape. Here it is shown that reference objects that are both spatially and spectrally separated from the object of interest can be used to obtain an approximation of the point spread function. The crucial observation, corroborated by analytical calculations, is that the spectrally shifted point spread function is strongly correlated to a spatially scaled one. With the approximate point spread function thus obtained, the speckle-like pattern is deconvolved to produce a clear and sharp image of the object on a speckle-like background of low intensity.
Wavefront shaping is increasingly being used in modern microscopy to obtain distortion-free, highresolution images deep inside inhomogeneous media. Wavefront shaping methods typically rely on the presence of a 'guidestar' in order to find the optimal wavefront to mitigate the scattering of light. However, this condition cannot be satisfied in most biomedical applications. Here, we introduce a novel, guidestar-free wavefront shaping method in which the optimal wavefront is computed using a digital model of the sample. The refractive index model of the sample, that serves as the input for the computation, is constructed in-situ by the microscope itself. In a proof of principle imaging experiment, we demonstrate a large improvement in the two-photon fluorescence signal through a diffuse medium, outperforming the state-of-the-art wavefront shaping techniques by a factor of 21.
Aqueous stability of sputter coated ZnO thin films were studied on two base materials, viz., polycarbonate (PC) and glass. The films showed higher stability on PC compared to glass, when exposed to aqueous buffered solution at pH-7.4, as studied by x-ray diffraction, surface reflectometry, and inductively coupled plasma-optical emission spectroscopy. Glucose oxidase (GOx) and cholesterol oxidase (Chl.Ox.) were used as model enzymes to study their electrochemical interaction with ZnO/PC. GOx showed a higher immobilization on ZnO/PC with an activity of 9.2 ± 1.7 mU cm compared to Chl.Ox. with an activity of 2.79 ± 0.5 mU cm. This is attributed to the larger crystallite size and higher Zn per unit area on PC as compared to glass which enabled a higher activity of GOx on ZnO/PC compared to ZnO/glass. Immobilization was mainly dependent on the surface residue and the charge of the enzyme as indicated by zeta potential which showed -23 mV for GOx compared to -6 mV for Chl.Ox. under physiological conditions. Further under unstirred condition, the reaction was limited by diffusion of the substrate for the enzyme. Chl.Ox. showed a lower activity as compared to GOx on the surface due to low diffusional coefficient of the bulky cholesterol molecule as compared to glucose. It was confirmed by low charge transfer resistance in electrochemical impedance spectroscopy for GOx (1.51 ± 0.072 × 10 Ω) as compared to Chl.Ox. (1.98 ± 0.09 × 10 Ω). But under stirring condition, the diffusion limitation was overcome, and the sensitivity for Chl.Ox./ZnO was 11.2 μA cmmM as compared to GOx/ZnO/PC with 3.5 μA cmmM. Thus, sputter coated ZnO thin films appeared to be good quality transducers for immobilization of oxidases with sensitivity dependent on the substrate diffusion and its potential application in biosensors.
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