This PDF file contains the front matter associated with SPIE Proceedings Volume 10333, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.

We demonstrate a 3D holographic tracking method to investigate particles motion in a microfluidic channel while unperturbed while inducing their migration through microfluidic manipulation. Digital holography (DH) in microscopy is a full-field, label-free imaging technique able to provide quantitative phase-contrast. The employed 3D tracking method is articulated in steps. First, the displacements along the optical axis are assessed by numerical refocusing criteria. In particular, an automatic refocusing method to recover the particles axial position is implemented employing a contrast-based refocusing criterion. Then, the transverse position of the in-focus object is evaluated through quantitative phase map segmentation methods and centroid-based 2D tracking strategy. The introduction of DH is thus suggested as a powerful approach for control of particles and biological samples manipulation, as well as a possible aid to precise design and implementation of advanced lab-on-chip microfluidic devices.

Abdominal aortic aneurysms (AAA) are a degenerative disease of the human aortic wall that may lead to weakening and eventually rupture of the wall with high mortality rates. Since the currently established criterion for surgical or endovascular treatment of the disease is imprecise in the individual case and treatment is not free of complications, the need for additional patient-individual biomarkers for short-term AAA rupture risk as basis for improved clinical decision making. Time resolved 3D ultrasound combined with speckle tracking algorithms is a novel non-invasive medical imaging technique that provides full-field displacement and strain measurements of aortic and aneurysmal wall motion. This is patient-individual information that has not been used so far to assess wall strength and rupture risk. The current study uses simple statistical indices of the heterogeneous spatial distribution of in-plane strain components as biomarkers for the pathological state of the aortic and aneurysmal wall. The pathophysiological rationale behind this approach are the known changes in microstructural composition of the aortic wall with progression of AAA development that results in increased stiffening and heterogeneity of the walls mechanical properties and in decreased wall strength. In a comparative analysis of the aortic wall motion of young volunteers without known cardiovascular diseases, aged arteriosclerotic patients without AAA, and AAA patients, mean values of all in-plane strain components were significantly reduced, and the heterogeneity of circumferential strain was significantly increased in the AAA group compared to both other groups. The capacity of the proposed method to differentiate between wall motion of aged, arteriosclerotic patients and AAA patients is a promising step towards a new method for in vivo assessment of AAA wall strength or stratification of AAA rupture risk as basis for improved clinical decision making on surgical or endovascular treatment of AAA.

A non-contact arterial-induced skin vibration inspection system is implemented. This optical metrology system is constructed with shadow Moiré configuration and the fringe analysis algorithm. Developed with the Region of Interested (ROI) capturing technique and the Two-dimensional Wavelet Transform (2D-CWT) method, this algorithm is able to retrieve the height-correlated phase information from the shadow Moiré fringe patterns. Using a commercial video camera or a CMOS image sensor, this system could monitor the skin-vibration induced by the cyclic deformation of inner layered artery. The cross-sectional variation and the rhythm of heart cycle could be continuously measured for health monitoring purposes. The average vibration amplitude of the artery at the wrist ranges between 20 μm and 50 μm, which is quite subtle comparing with the skin surface structure. Having the non-stationary motion of human body, the traditional phase shifting (PS) technique can be very unstable due to the requirement of several frames of images, especially for case that artery is continuously pumping. To bypass this fundamental issue, the shadow Moiré technique is introduced to enhance the surface deformation characteristic. And the phase information is retrieved by the means of spectrum filtering instead of PS technique, which the phase is calculated from intensity maps of multiple images. The instantaneous surface can therefore be reconstructed individually from each frame, enabling the subtle arterial-induced skin vibration measurement. The comparative results of phase reconstruction between different fringe analysis algorithms will be demonstrated numerically and experimentally. And the electrocardiography (ECG) results will used as the reference for the validity of health monitoring potential of the non-contact arterial-induced skin vibration inspection system.

We present an experimental comparison of optical tomography techniques: Learning Tomography (LT), an iterative method based on beam propagation model and diffraction tomography (DT) based Born and Rytov approximation. Imaging experiments were performed on yeast cells clusters fixed in a transparent agarose gel. In particular, we compare the LT with a similar iterative but linear method based on Rytov approximation.

Quality and reproducibility of cell-based assays strongly depend on the quality of the underlying cell culture which is influenced by various parameters like nutrient and growth factor availability, buffer conditions, subculture routines and optimal cell concentrations. Thus, methods for accurate assessment of objective cell parameters that characterize a specific cell line and detect global changes in cell culture are highly desirable. During the past years, quantitative phase imaging has been recognized as a promising tool for quantitative label-free live cell analysis. We demonstrate the utilization of quantitative phase imaging with digital holographic microscopy (DHM) to quantify the impact of cell culture conditions on single cells using a pancreatic tumor cell model. Label-free quantitative phase imaging of detached cells in suspension is performed by Michelson interferometer-based self-interference DHM. The quantitative phase images of the cells are analyzed for refractive index, volume and dry mass. We show that the evaluation of quantitative DHM phase images allows to extract absolute biophysical cellular parameters that are related to cell layer confluence states. In summary, the results of our study demonstrate that DHM is capable for label-free imaging cytometry with novel biophysical data sets that are acquired with minimum sample preparation for sophisticated monitoring of cell morphology alterations that are related to changes of cell culture conditions.

Cell of human blood stream are divided into two groups: Red Blood Cells (RBC) and White Blood Cells (WBC). RBC have a peculiar biconcave disk shape and they are responsible for the delivering of O₂ and CO₂ through the body. WBC are a more widespread class of cell ensuring immunity against pathogens. They can be divided in two main classes: granulocyte cells and A-granulocyte cells. Neutrophils, basophils and eosinophils belong to the granulocyte cell class, while lymphocytes and monocytes belong to A-granulocyte. Both in RBC and WBC, the intrinsic physical properties of a cell are indicators of cell condition and, furthermore, of the overall human body state. Thus, the accurate comprehension of the physiological structure of WBCs is fundamental to recognize diseases. Here we show the possibility to simple and straightforwardly characterize the physical properties of individual RBC and mononuclear WBC in a microfluidic context, using a wide angle light scattering apparatus and a corresponding theoretical simulation of Optical Signature (OS). A non-Newtonian polymer alignment solution for cell is used to ensure an individual cell alignment in the microfluidic flow, thus permitting a precise investigation. Additionally, Quantitative Phase Imaging (QPI) holographic measurements are performed to estimate cell morphometric features, such as their refractive index. We analyzed more than 200 WBCs and 100 RBCs of three different probands. Results showed distinct cell populations according to their measured dimensions and shape, which can be associated to the presence of RBC, lymphocytes and monocytes.

Optical Microscopy has been applied to life science from its birth and reached widespread application due to its major advantages: limited perturbation of the biological tissue and the easy accessibility of the light sources. However, as the spatial and time resolution requirements and the time stability of the microscopes increase, researchers are struggling against some of its limitations: limited transparency and the refractivity of the living tissue to light and the field perturbations induced by the path in the tissue. We have developed a compact stand-alone, completely scan-less, optical setup that allows to acquire non-linear excitation images and to measure the sample dynamics simultaneously on an ensemble of arbitrary chosen regions of interests. The image is obtained by shining a square array of spots on the sample obtained by a spatial light modulator and by shifting it (10 ms refresh time) on the sample. The final image is computed from the superposition of (100-1000) images. Filtering procedures can be applied to the raw images of the excitation array before building the image. We discuss results that show how this setup can be used for the correction of wave front aberrations induced by turbid samples (such as living tissues) and for the computation of space-time cross-correlations in complex networks.

Adaptive optics (AO)-enhanced imaging of the retina is now reaching a level of technical maturity which fosters its expanding use in research and clinical centers in the world. By achieving wavelength-limited resolution it did not only allow a better observation of retinal substructures already visible by other means, it also broke anatomical frontiers such as individual photoreceptors or vessel walls. The clinical applications of AO-enhanced imaging has been slower than that of optical coherence tomography because of the combination of technical complexity, costs and the paucity of interpretative scheme of complex data. In several diseases, AO-enhanced imaging has already proven to provide added clinical value and quantitative biomarkers. Here, we will review some of the clinical applications of AO-enhanced en face imaging, and trace perspectives to improve its clinical pertinence in these applications. An interesting perspective is to document cell motion through time-lapse imaging such as during agerelated macular degeneration. In arterial hypertension, the possibility to measure parietal thickness and perform fine morphometric analysis is of interest for monitoring patients. In the near future, implementation of novel approaches and multimodal imaging, including in particular optical coherence tomography, will undoubtedly expand our imaging capabilities. Tackling the technical, scientific and medical challenges offered by high resolution imaging are likely to contribute to our rethinking of many retinal diseases, and, most importantly, may find applications in other areas of medicine.

The complete phase and amplitude information of biological specimens can be easily determined by phase-shifting digital holography. Spatial light modulators (SLMs) based on liquid crystal technology, with a frame-rate around 60 Hz, have been employed in digital holography. In contrast, digital micro-mirror devices (DMDs) can reach frame rates up to 22 kHz. A method proposed by Lee to design computer generated holograms (CGHs) permits the use of such binary amplitude modulators as phase-modulation devices. Single-pixel imaging techniques record images by sampling the object with a sequence of micro-structured light patterns and using a simple photodetector. Our group has reported some approaches combining single-pixel imaging and phase-shifting digital holography. In this communication, we review these techniques and present the possibility of a high-speed single-pixel phase-shifting digital holography system with phase-encoded illumination. This system is based on a Mach-Zehnder interferometer, with a DMD acting as the modulator for projecting the sampling patterns on the object and also being used for phase-shifting. The proposed sampling functions are phaseencoded Hadamard patterns generated through a Lee hologram approach. The method allows the recording of the complex amplitude distribution of an object at high speed on account of the high frame rates of the DMD. Reconstruction may take just a few seconds. Besides, the optical setup is envisaged as a true adaptive system, which is able to measure the aberration induced by the optical system in the absence of a sample object, and then to compensate the wavefront in the phasemodulation stage.

Holographic techniques have been used to measure the shape and the radial deformation of a blood vessel model and a real sheep aorta. Measurements are obtained from several holograms recorded for different object states. For each object state, two holograms with two different wavelengths are multiplexed in the same digital recording. Thus both holograms are simultaneously recorded but the information from each of them is separately obtained. The shape analysis gives a wrapped phase map whose fringes are related to a synthetic wavelength. After a filtering and unwrapping process, the 3D shape can be obtained. The shape data for each line are fitted to a circumference in order to determine the local vessel radius and center. The deformation analysis also results in a wrapped phase map, but the fringes are related to the laser wavelength used in the corresponding hologram. After the filtering and unwrapping process, a 2D map of the deformation in an out-of-plane direction is reconstructed. The radial deformation is then calculated by using the shape information.

The simultaneous presence of the real and virtual images in the hologram reconstruction is inherent in the in-line holography. This drawback can be overcome with a shifted knife-edge aperture at the focal plane of the imaging lens. The shifted aperture DIH produces holograms where the real and virtual images are completely separated. In this paper we propose a modification of the shifted aperture DIH that allows recording two holograms simultaneously using one camera, while retaining the simplicity of the in-line configuration and the advantage of the shifted-aperture strategy. As in typical stereoscopy, the advantage of this configuration is limited by the angle between the two illuminating beams, and therefore the aperture size. Some improvement on the out-of-plane resolution can be expected from a combined analysis of the multiplexed holograms. In order to compare this technique with other in-line holographic configurations, several experiments have been performed to study the spatial resolution along the optical axis. The capabilities of the different techniques for characterizing the flow in a flexible and transparent model of a carotid bifurcation are also investigated.

Superresolution image reconstruction method based on the structured illumination microscopy (SIM) principle with reduced and simplified pattern set is presented. The method described needs only 2 sinusoidal patterns shifted by half a period for each spatial direction of reconstruction, instead of the minimum of 3 for the previously known methods. The method is based on estimating redundant frequency components in the acquired set of modulated images. Digital processing is based on linear operations. When applied to several spatial orientations, the image set can be further reduced to a single pattern for each spatial orientation, complemented by a single non-modulated image for all the orientations. By utilizing this method for the case of two spatial orientations, the total input image set is reduced up to 3 images, providing up to 2-fold improvement in data acquisition time compared to the conventional 3-pattern SIM method. Using the simplified pattern design, the field of view can be doubled with the same number of spatial light modulator raster elements, resulting in a total 4-fold increase in the space-time product. The method requires precise knowledge of the optical transfer function (OTF). The key limitation is the thickness of object layer that scatters or emits light, which requires to be sufficiently small relatively to the lens depth of field. Numerical simulations and experimental results are presented. Experimental results are obtained on the SIM setup with the spatial light modulator based on the 1920x1080 digital micromirror device.

The applications of fluorescence microscopy span medical diagnostics, bioengineering and biomaterial analytics. Full exploitation of fluorescent microscopy is hampered by imperfections in illumination, detection and filtering. Mainly, errors stem from deviations induced by real-world components inducing spatial or angular variations of propagation properties along the optical path, and they can be addressed through consistent and accurate calibration. For many applications, uniform signal to noise ratio (SNR) over the imaging area is required. Homogeneous SNR can be achieved by quantifying and compensating for the signal bias. We present a method to quantitatively characterize novel reference materials as a calibration reference for biomaterials analytics. The reference materials under investigation comprise thin layers of fluorophores embedded in polymer matrices. These layers are highly homogeneous in their fluorescence response, where cumulative variations do not exceed 1% over the field of view (1.5 x 1.1 mm). An automated and reproducible measurement methodology, enabling sufficient correction for measurement artefacts, is reported. The measurement setup is equipped with an autofocus system, ensuring that the measured film quality is not artificially increased by out-of-focus reduction of the system modulation transfer function. The quantitative characterization method is suitable for analysis of modified bio-materials, especially through patterned protein decoration. The imaging method presented here can be used to statistically analyze protein patterns, thereby increasing both precision and throughput. Further, the method can be developed to include a reference emitter and detector pair on the image surface of the reference object, in order to provide traceable measurements.

It has been widely proven in literature that most optical microscopy techniques can greatly benefit from the application of adaptive optics correction of phase aberrations through an adaptive optical element, such as a deformable mirror or a spatial light modulator. However, adaptive optics is not yet widely adopted in the life sciences community, mostly due to the lack of adaptive commercial microscopy systems, and the inherent technical difficulty in modifying an existing microscopy setup to integrate an adaptive element, both on the software and hardware sides. We present a plug-and-play adaptive optics module for generic optical microscopes, based on a prototype refractive 18 actuators adaptive optical element, which can be inserted in any microscope between the objective and the microscope body. Correction is performed in a sensorless fashion, optimizing image quality metrics of the image presented to the user on screen. The results presented show how an end-user oriented commercial confocal laser scanning microscope (Leica SP5) can be upgraded with adaptive optics with minor hardware modifications, and no changes to the microscope control software.

The Digital Holographic Microscopy in Transmission technique (DHM) is considered a useful tool in the noninvasive quantifying of transparent biological objects like living cells. In this work, we propose this technique to study and to monitor control macrophages infected by Leishmania (mouse lineJ774.A1). When the promastigotes enter in contact with healthy macrophages, they got phagocytosed and latterly confined in the formed parasitophorous vacuole. These processes change the morphology and density of the host macrophage. Both parameters can be measured in a label-free analysis of cells with the aid of the DHM technique. Our technique begins with the optical record of the holograms using a modified Mach-Zehnder interferometer and the reconstruction of the complex optical field transmitted by macrophages. In the latter point, we employ the angular spectrum algorithm. With the complex optical field reconstruction, we compute the field amplitude and the phase difference maps, which leads to describe one morphological characterization for the samples. Using phase difference maps is possible to measure internal variations for the integral refractive index, estimating the infection level of macrophages. Through the changes in the integral refractive index, it is also possible to describe and quantify in two different states the evolution of the infection. With these results some parameters of cells have been quantified, making the DHM technique a viable tool for diagnosis of biological samples under the presence of some pathogen.

Spatial light modulators (SLM) are currently used in many applications in optical microscopy and imaging. One of the most promising methods is the use of liquid crystal displays (LCD) as programmable phase diffractive optical elements (DOE) placed in the Fourier plane giving access to the spatial frequencies which can be phased shifted individually, allowing to emulate a wealth of contrast enhancing methods for both amplitude and phase samples. We use phase and polarization modulation of LCD to implement an on-axis microscope optical system. The LCD used are Hamamatsu liquid crystal on silicon (LCOS) SLM free of flicker, thus showing a full profit of the SLM space bandwidth, as opposed to optical systems in the literature forced to work off-axis due to the strong zero-order component. Taking benefits of the phase modulation of the LCOS we have implemented different microscopic imaging operations, such as high-pass and low-pass filtering in parallel using programmable blazed gratings. Moreover, we are able to control polarization modulation to display two orthogonal linear state of polarization images than can be subtracted or added by changing the period of the blazed grating. In that sense, Differential Interference Contrast (DIC) microscopy can be easily done by generating two images exploiting the polarization splitting properties when a blazed grating is displayed in the SLM. Biological microscopy samples are also used.

Here we introduce a compact holographic microscope embedded onboard a Lab-on-a-Chip (LoC) platform. A wavefront division interferometer is realized by writing a polymer grating onto the channel to extract a reference wave from the object wave impinging the LoC. A portion of the beam reaches the samples flowing along the channel path, carrying their information content to the recording device, while one of the diffraction orders from the grating acts as an off-axis reference wave. Polymeric micro-lenses are delivered forward the chip by Pyro-ElectroHydroDynamic (Pyro-EHD) inkjet printing techniques. Thus, all the required optical components are embedded onboard a pocket device, and fast, non-iterative, reconstruction algorithms can be used. We use our device in combination with a novel high-throughput technique, named Space-Time Digital Holography (STDH). STDH exploits the samples motion inside microfluidic channels to obtain a synthetic hologram, mapped in a hybrid space-time domain, and with intrinsic useful features. Indeed, a single Linear Sensor Array (LSA) is sufficient to build up a synthetic representation of the entire experiment (i.e. the STDH) with unlimited Field of View (FoV) along the scanning direction, independently from the magnification factor. The throughput of the imaging system is dramatically increased as STDH provides unlimited FoV, refocusable imaging of samples inside the liquid volume with no need for hologram stitching. To test our embedded STDH microscopy module, we counted, imaged and tracked in 3D with high-throughput red blood cells moving inside the channel volume under non ideal flow conditions.

The determination of contact area is pivotal to understand how biomaterials properties influence cell adhesion. In particular, the influence of surface charges is well-known but still controversial, especially when new functional materials and methods are introduced. Here, we use for the first time Holographic Total Internal Reflection Microscopy (HoloTIRM) to study the influence of the spontaneous polarization of ferroelectric lithium niobate (LN) on the adhesion properties of fibroblast cells. The selective illumination of a very thin region directly above the substrate, achieved by Total Internal Reflection, provides high-contrast images of the contact regions. Holographic recording, on the other hand, allows for label-free quantitative phase imaging of the contact areas between cells and LN. Phase signal is more sensitive in the first 100nm and, thus more reliable in order to locate focal contacts. This work shows that cells adhering on negatively polarized LN present a significant increase of the contact area in comparison with cells adhering on the positively polarized LN substrate, as well as an intensification of contact vicinity. This confirms the potential of LN as a platform for investigating the role of charges on cellular processes. The similarity of cell adhesion behavior on negatively polarized LN and glass control also confirms the possibility to use LN as an active substrate without impairing cell behavior.

Plenoptic Imaging (PI) is a novel optical technique for achieving tridimensional imaging in a single shot. In conventional PI, a microlens array is inserted in the native image plane and the sensor array is moved behind the microlenses. On the one hand, the microlenses act as imaging pixels to reproduce the image of the scene; on the other hand, each microlens reproduces on the sensor array an image of the camera lens, thus providing the angular information associated with each imaging pixel. The recorded propagation direction is exploited, in post- processing, to computationally retrace the geometrical light path, thus enabling the refocusing of different planes within the scene, the extension of the depth of field of the acquired image, as well as the 3D reconstruction of the scene. However, a trade-off between spatial and angular resolution is built in the standard plenoptic imaging process. We demonstrate that the second-order spatio-temporal correlation properties of light can be exploited to overcome this fundamental limitation. Using two correlated beams, from either a chaotic or an entangled photon source, we can perform imaging in one arm and simultaneously obtain the angular information in the other arm. In fact, we show that the second order correlation function possesses plenoptic imaging properties (i.e., it encodes both spatial and angular information), and is thus characterized by a key re-focusing and 3D imaging capability. From a fundamental standpoint, the plenoptic application is the first situation where the counterintuitive properties of correlated systems are effectively used to beat intrinsic limits of standard imaging systems. From a practical standpoint, our protocol can dramatically enhance the potentials of PI, paving the way towards its promising applications.

High-throughput single-cell analysis is a challenging target for implementing advanced biomedical applications. An excellent candidate for this aim is label-free tomographic phase microscopy. However, in-line tomography is very difficult to be implemented in practice, as it requires complex setup for rotating the sample and/or illuminate the cell along numerous directions [1]. We exploit random rolling of cells while they are flowing along a microfluidic channel demonstrating that it is possible to obtain in-line phase-contrast tomography by adopting strategies for intelligent wavefronts analysis thus obtaining complete retrieval of both 3D-position and orientation of rotating cells [2]. Thus, by numerical wavefront analysis a-priori knowledge of such information is no longer needed. This approach makes continuos-flow cyto-tomography suitable for practical operation in real-world, single-cell analysis and with substantial simplification of the optical system avoiding any mechanical/optical scanning of light source. Demonstration is given for different classes of biosamples, red-blood-cells (RBCs), diatom algae and fibroblast cells [3]. Accurate characterization of each type of cells is reported despite their very different nature and materials content, thus showing the proposed method can be extended, by adopting two alternate strategies of wavefront-analysis, to many classes of cells. In particular, for RBCs we furnish important parameters as 3D morphology, Corpuscular Hemoglobin (CH), Volume (V), and refractive index (RI) for each single cell in the total population [3]. This could open a new route in blood disease diagnosis, for example for the isolation and characterization of "foreign" cells in the blood stream, the so called Circulating Tumor Cells (CTC), early manifestation of metastasis.

A critical factor of endoprostheses is the quality of the tribological pairing. The objective of this research project is to manufacture stochastically porous aluminum oxide surface coatings with high wear resistance and an active friction minimization. There are many experimental and computational techniques from mercury porosimetry to imaging methods for studying porous materials, however, the characterization of disordered pore networks is still a great challenge. To meet this challenge it is striven to gain a three dimensional high resolution reconstruction of the surface. In this work, the reconstruction is approached by repeatedly milling down the surface by a fixed decrement while measuring each layer using a confocal laser scanning microscope (CLSM). The so acquired depth data of the successive layers is then registered pairwise. Within this work a direct registration approach is deployed and implemented in two steps, a coarse and a fine alignment. The coarse alignment of the depth data is limited to a translational shift which occurs in horizontal direction due to placing the sample in turns under the CLSM and the milling machine and in vertical direction due to the milling process itself. The shift is determined by an approach utilizing 3D phase correlation. The fine alignment is implemented by the Trimmed Iterative Closest Point algorithm, matching the most likely common pixels roughly specified by an estimated overlap rate. With the presented two-step approach a proper 3D registration of the successive depth data of the layer is obtained.

Developing the recently discovered concept of RBCs as microlenses, we demonstrate further applications in wavefront analysis and diagnostics. Correlation between RBC’s morphology and its behavior as a refractive optical element has been established. In fact, any deviation from the healthy RBC morphology can be seen as additional aberration in the optical wavefront passing through the cell. By this concept, accurate localization of focal spots of RBCs can become very useful in blood disorders identification. Moreover, By modelling RBC as bio-lenses through Zernike polynomials it is possible to identify a series of orthogonal parameters able to recognise RBC shapes. The main improvement concerns the possibility to combine such parameters because of their independence conversely to standard image-based analysis where morphological factors are dependent each-others. We investigate the three-dimensional positioning of such focal spots over time for samples with two different osmolarity conditions, i.e. discocytes and spherocytes. Finally, Zernike polynomials wavefront analysis allows us to study the optical behavior of RBCs under an optically-induced mechanical stress. Detailed wavefront analysis provides comprehensive information about the aberrations induced by the deformation obtained using optical tweezers. This could open new routes for analyzing cell elasticity by examining optical parameters instead of direct but with low resolution strain analysis, thanks to the high sensitivity of the interferometric tool.

New approach to perform a real-time biochemical component detection based on simultaneous analysis of spectral changes of whispering gallery modes (WGM) and fluorescence markers used for biochemical components tagging. Microcavity array sensor was chosen as detection unit. Experimental data on detection of bovine serum albumin protein solution using both techniques simultaneously is represented.

When using Shack-Hartmann wavefront sensors (SH) and Zernike coefficients (Zs) in applications where the position of the measurement and the point of interest are far apart, as it is common practice in ophthalmic optics, problems in the interpretation of the values of the Zs arise, related to how the shape of the wavefront propagates along the beam. One typical example is pupil conjugation where an auxiliary lens is added to match the size of the area of the interest of the beam with the size of the entrance pupil of the SH used for measurements. In the present work, we address this problem in the framework of a numerical scheme for modeling the beam propagation. We calculate the wavefronts with exact ray tracing plus the fitting of the impacts so as to match a rectangular grid. This procedure allows the subsequent calculation of the Zs or, similarly, the pupil function at an arbitrary plane perpendicular to the optical axis. All the numerical methods and procedures have been implemented in MATLAB code and can be illustrated by running the MATLAB script for the setup configuration that is being considered. Several examples are presented to illustrate the previous ideas and to show the real capabilities of our procedures. They will help to clarify the issues actually found in practical setups for beam manipulation, often encountered in ophthalmic optics.

The phenomenon of inclusions or microvacuoles in intraocular lenses (IOL), often referred to glistenings due to their appearance when visualized in slit-lamp exams, is main cause of decreased visual in people after IOL implantation. For this reason, there is a huge request by the market of new polymers able to reduce, or even eliminate, the formation of such microvacuoles. In such frame, the use of advanced optical techniques, able to provide a deeper insight on the glistering formation, is strongly required. In particular, Raman spectroscopy (RS) is ideally suited for the analysis of polymers, due to its well-know sensitivity to highly polarizable chemical groups, commonly found in the polymer chains backbones. Moreover, the combination of RS with optical microscopy (Raman micro-spectroscopy) paves the way for real, information-rich chemical mapping of polymeric materials (Raman imaging). In this paper, we analyze the formation of microvacuoles in IOLs following a thermal treatment. In particular, we performed a chemical mapping of a single microvacuole, which allowed us to infer on its effective chemical composition. In order to investigate on the reversibility of glistenings formation, this analysis was repeated as function of time after thermal treatment, in different IOL environments. It turns out that this phenomenon is partially reversible, with an almost complete disappearance of microvacuoles in a dry environment.

Recently, terahertz (THz) photonic crystal waveguides based on sapphire shaped crystals have been proposed. These waveguides combine unique properties of sapphire with advantages of the edge-defined film-fed growth (EFG) or Stepanov technique of shaped crystal growth and allow guiding THz waves in a wide spectral range with small dispersion and losses. The sapphire photonic crystal waveguides are capable for operation in aggressive environment, which makes possible to perform high-temperature and high-pressure THz measurements, as well as THz measurements of aggressive chemicals. In this paper, the technological aspects of sapphire THz photonic crystal waveguide manufacturing by the EFG/Stepanov technique (including, the problems of seeding and automated control of multichannel shaped crystal growth) have been described. Prospective applications of sapphire photonic crystal waveguides in various branches of THz science and technology have been discussed.

Design of approaches and methods of the oncological diseases diagnostics has special significance. It allows determining any kind of tumors at early stages. The development of optical and laser technologies provided increase of a number of methods allowing making diagnostic studies of oncological diseases. A promising area of biomedical diagnostics is the development of automated nondestructive testing systems for the study of the skin polarizing properties based on backscattered radiation detection. Specification of the examined tissue polarizing properties allows studying of structural properties change influenced by various pathologies. Consequently, measurement and analysis of the polarizing properties of the scattered optical radiation for the development of methods for diagnosis and imaging of skin in vivo appear relevant. The purpose of this research is to design the algorithm of photons migration in the multilayer skin structure. In this research, the algorithm of photons migration in the multilayer skin structure was designed. It is based on the use of the Monte Carlo method. Implemented Monte Carlo method appears as a tracking the paths of photons experiencing random discrete direction changes before they are released from the analyzed area or decrease their intensity to negligible levels. Modeling algorithm consists of the medium and the source characteristics generation, a photon generating considering spatial coordinates of the polar and azimuthal angles, the photon weight reduction calculating due to specular and diffuse reflection, the photon mean free path definition, the photon motion direction angle definition as a result of random scattering with a Henyey-Greenstein phase function, the medium’s absorption calculation. Biological tissue is modeled as a homogeneous scattering sheet characterized by absorption, a scattering and anisotropy coefficients.

The main aspect in developing methods for optical diagnostics and visualization of biological tissues using polarized radiation is the transformation analysis of the state of light polarization when it is scattered by the medium. The polarization characteristic spatial distributions of the detected scattered radiation, in particular the degree of depolarization, have a pronounced anisotropy. The presence of optical anisotropy can provide valuable additional information on the structural features of the biological object and its physiological status. Analysis of the polarization characteristics of the scattered radiation of biological tissues in some cases provides a qualitatively new results in the study of biological samples. These results can be used in medicine and food industry.

Our study focuses on an analysis of the original method of investigation biological tissues in the spectral OCT (optical coherence tomography) with usage hyperchromatic lenses. Using hyperchromatic lens, i.e. the lens with uncorrected longitudinal color allows scanning in the depth of the object by changing the wavelength of the emitter. In this case, the depth of the scan will be determined not by the microlens depth of field, but the value of axial color. In our study, we demonstrated the advantages of this method of research on biological tissues existing. Spectral OCT schemes with the hyperchromatic lens could increase the depth of spectral scanning, eliminate the use of multi-channel systems with a set of microscope objectives, reduce the time of measurement. In our paper, we show the developed method of calculation of hyperchromatic lenses and hybrid hyperchromatic lens consisting of a diffractive and refractive component in spectral OCT systems. We also demonstrate the results of aberration calculation designed microscope lenses. We show examples of developed hyperchromatic lenses with the diffractive element and without it.

In this paper, we propose a generalization of the box fractal dimension in images by considering the curve obtained from its value as a function of the binarization threshold. This curve can be used to describe speckle patterns. We show some examples of both objective simulated and experimental and subjective speckle in some cases of interest. The concept can be extended for all types of images.

Nanoparticles with second-order optical nonlinearities have been widely studied because of their applications in clinical and industrial applications. The nonlinear absorption coefficient is an important parameter which should be determined in order to fully grasp the implications of these materials. In this work Moire deflectometry method is used to measure the nonlinear absorption coefficient of nanoparticles. In the proposed method the divergence of beam is measured instead of measuring the intensity of divergent ray. Two beams are used; one is a comparatively high intensity laser beam which is used as the interacting beam and the other is used as probe, wide beam with low intensity that is radiated to the first beam vertically. By analyzing the probe beam using Moiré deflectometry, the nonlinear absorption coefficient is measured.

Aberration correction was one of the most important and challenging parts of optical trapping systems. Many works try to overcome aberration which is due to trapping setup itself or refractive index mismatch of glass slab and nanoparticles solutions. In this article, we investigate a new method for aberration correction based on a nonlinear refractive index of materials. With the emitting laser to the trapped particle, the refractive index of the trapping site increases and as a result, its aberration reduce. In accordance, this method offers a simple and low-cost way for increasing trapping stiffness.

Peculiarities of optical design for optical coherence tomography (OCT) system with illumination by a swept-source in the spectral range 1.26-1.36 μm are considered. In the OCT system, an object is illuminated by light intensity distribution in the form of line providing high power efficiency of the light source when evaluating micro structure of objects. A linearray photo detector with the frame acquisition rate of a few tens of kilohertz is utilized that allows obtaining B-scans without mechanical lateral scanning. The illumination power density at each point of investigated object is much less with respect to conventional "flying spot" methods that is important when studying biological objects not resistant to intensive light. Results of experimental investigations utilizing the Linnik micro interferometer optical scheme are given. Experimental tomograms of different objects are presented.

The development of tools for rapid food quality inspection is a highly pursued goal. These could be valuable devices to be used by food producers in factories or the customers themselves in specific installations at the marketplace. Here we show how speckle patterns in coherent imaging systems can be can be employed as indicators of the presence of bacteria colonies contaminating food or water. Speckle decorrelation is induced by the self-propelling movement of these organisms when they interact with coherent light. Hence, their presence can be detected using a simple setup in a condition in which the single element cannot be imaged, but the properties of the ensemble can be exploited. Thanks to the small magnification factor we set, our system can inspect a large Field-of-View (FoV). We show the possibility to discriminate between fresh and contaminated food, thus paving the way to the rapid food quality testing by consumers at the marketplace.

In this paper, we propose digital holography in transmission configuration as an effective method to measure the time-dependent thickness of polymeric films during bubble blowing. We designed a complete set of experiments to measure bubble thickness, including the evaluation of the refractive index of the polymer solution. We report the measurement of thickness distribution along the film during the bubble formation process until the bubble‘s rupture. Based on those data, the variation range and variation trend of bubble film thickness are clearly measured during the process of expansion to fracture is indicated.