High resolution multiple excitation spot optical microscopy

We propose fundamental improvements in three-dimensional (3D) resolution of multiple excitation spot optical microscopy. The excitation point spread function (PSF) is generated by two interfering counter-propagating depth-of-focus beams along the optical axis. Detection PSF is obtained by coherently interfering the emitted fluorescent light (collected by both the objectives) at the detector. System PSF shows upto 14-fold reduction in focal volume as compared to confocal, and almost 2-fold improvement in lateral resolution. Proposed PSF has the ability to simultaneously excite multiple 3D-spots of sub-femtoliter volume. Potential applications are in fluorescence microscopy and nanobioimaging. Copyright 2011 Author(s). This article is distributed under a Creative Commons Attribution 3.0 Unported License. [doi:10.1063/1.3598413]

Multiple excitation nano-spot generation and confocal detection for far-field microscopy

An imaging technique is developed for the controlled generation of multiple excitation nano-spots for far-field microscopy. The system point spread function (PSF) is obtained by interfering two counter-propagating extended depth-of-focus PSF (DoF-PSF), resulting in highly localized multiple excitation spots along the optical axis. The technique permits (1) simultaneous excitation of multiple planes in the specimen; (2) control of the number of spots by confocal detection; and (3) overcoming the point-by-point based excitation. Fluorescence detection from the excitation spots can be efficiently achieved by Z-scanning the detector/pinhole assembly. The technique complements most of the bioimaging techniques and may find potential application in high resolution fluorescence microscopy and nanoscale imaging.

Chain of excitation spot generation by aperture engineering

An optical technique is proposed for obtaining multiple excitation spots. Phase-matched counter propagating extended depth-of-focus fields were superimposed along the optical axis for generating multiple localized excitation spots. Moreover, the filtering effect due to the optical mask increases the lateral resolution. Proposed technique introduces the concept of simultaneous multiplane excitation and improves three-dimensional resolution. (C) 2010 American Institute of Physics.

Simultaneous multilayer scanning and detection for multiphoton fluorescence microscopy

Fast three-dimensional (3D) imaging requires parallel optical slicing of a specimen with an efficient detection scheme. The generation of multiple localized dot-like excitation structures solves the problem of simultaneous slicing multiple specimen layers, but an efficient detection scheme is necessary. Confocal theta detection (detection at 90 degrees to the optical axis) provides a suitable detection platform that is capable of cross-talk-free fluorescence detection from each nanodot (axial dimension approximate to 150 nm). Additionally, this technique has the unique feature of imaging a specimen at a large working distance with super-resolution capabilities. Polarization studies show distinct field structures for fixed and fluid samples, indicating a non-negligible field-dipole interaction. The realization of the proposed imaging technique will advance and diversify multiphoton fluorescence microscopy for numerous applications in nanobioimaging and optical engineering.

Singular value decomposition based computationally efficient algorithm for rapid dynamic near-infrared diffuse optical tomography

Purpose: A computationally efficient algorithm (linear iterative type) based on singular value decomposition (SVD) of the Jacobian has been developed that can be used in rapid dynamic near-infrared (NIR) diffuse optical tomography. Methods: Numerical and experimental studies have been conducted to prove the computational efficacy of this SVD-based algorithm over conventional optical image reconstruction algorithms. Results: These studies indicate that the performance of linear iterative algorithms in terms of contrast recovery (quantitation of optical images) is better compared to nonlinear iterative (conventional) algorithms, provided the initial guess is close to the actual solution. The nonlinear algorithms can provide better quality images compared to the linear iterative type algorithms. Moreover, the analytical and numerical equivalence of the SVD-based algorithm to linear iterative algorithms was also established as a part of this work. It is also demonstrated that the SVD-based image reconstruction typically requires O(NN2) operations per iteration, as contrasted with linear and nonlinear iterative methods that, respectively, requir O(NN3) and O(NN6) operations, with ``NN'' being the number of unknown parameters in the optical image reconstruction procedure. Conclusions: This SVD-based computationally efficient algorithm can make the integration of image reconstruction procedure with the data acquisition feasible, in turn making the rapid dynamic NIR tomography viable in the clinic to continuously monitor hemodynamic changes in the tissue pathophysiology.

Emitter near an arbitrary body: Purcell effect, optical theorem and the Wheeler-Feynman absorber

The altered spontaneous emission of an emitter near an arbitrary body can be elucidated using an energy balance of the electromagnetic field. From a classical point of view it is trivial to show that the field scattered back from any body should alter the emission of the source. But it is not at all apparent that the total radiative and non-radiative decay in an arbitrary body can add to the vacuum decay rate of the emitter (i.e.) an increase of emission that is just as much as the body absorbs and radiates in all directions. This gives us an opportunity to revisit two other elegant classical ideas of the past, the optical theorem and the Wheeler-Feynman absorber theory of radiation. It also provides us alternative perspectives of Purcell effect and generalizes many of its manifestations, both enhancement and inhibition of emission. When the optical density of states of a body or a material is difficult to resolve (in a complex geometry or a highly inhomogeneous volume) such a generalization offers new directions to solutions. (c) 2012 Elsevier Ltd. All rights reserved.

Mesh Simplification Based on Edge Collapsing Could Improve Computational Efficiency in Near Infrared Optical Tomographic Imaging

The diffusion equation-based modeling of near infrared light propagation in tissue is achieved by using finite-element mesh for imaging real-tissue types, such as breast and brain. The finite-element mesh size (number of nodes) dictates the parameter space in the optical tomographic imaging. Most commonly used finite-element meshing algorithms do not provide the flexibility of distinct nodal spacing in different regions of imaging domain to take the sensitivity of the problem into consideration. This study aims to present a computationally efficient mesh simplification method that can be used as a preprocessing step to iterative image reconstruction, where the finite-element mesh is simplified by using an edge collapsing algorithm to reduce the parameter space at regions where the sensitivity of the problem is relatively low. It is shown, using simulations and experimental phantom data for simple meshes/domains, that a significant reduction in parameter space could be achieved without compromising on the reconstructed image quality. The maximum errors observed by using the simplified meshes were less than 0.27% in the forward problem and 5% for inverse problem.

Data-resolution based optimization of the data-collection strategy for near infrared diffuse optical tomography

Purpose: To optimize the data-collection strategy for diffuse optical tomography and to obtain a set of independent measurements among the total measurements using the model based data-resolution matrix characteristics. Methods: The data-resolution matrix is computed based on the sensitivity matrix and the regularization scheme used in the reconstruction procedure by matching the predicted data with the actual one. The diagonal values of data-resolution matrix show the importance of a particular measurement and the magnitude of off-diagonal entries shows the dependence among measurements. Based on the closeness of diagonal value magnitude to off-diagonal entries, the independent measurements choice is made. The reconstruction results obtained using all measurements were compared to the ones obtained using only independent measurements in both numerical and experimental phantom cases. The traditional singular value analysis was also performed to compare the results obtained using the proposed method. Results: The results indicate that choosing only independent measurements based on data-resolution matrix characteristics for the image reconstruction does not compromise the reconstructed image quality significantly, in turn reduces the data-collection time associated with the procedure. When the same number of measurements (equivalent to independent ones) are chosen at random, the reconstruction results were having poor quality with major boundary artifacts. The number of independent measurements obtained using data-resolution matrix analysis is much higher compared to that obtained using the singular value analysis. Conclusions: The data-resolution matrix analysis is able to provide the high level of optimization needed for effective data-collection in diffuse optical imaging. The analysis itself is independent of noise characteristics in the data, resulting in an universal framework to characterize and optimize a given data-collection strategy. (C) 2012 American Association of Physicists in Medicine. http://dx.doi.org/10.1118/1.4736820]

Effective contrast recovery in rapid dynamic near-infrared diffuse optical tomography using l(1)-norm-based linear image reconstruction method

Traditional image reconstruction methods in rapid dynamic diffuse optical tomography employ l(2)-norm-based regularization, which is known to remove the high-frequency components in the reconstructed images and make them appear smooth. The contrast recovery in these type of methods is typically dependent on the iterative nature of method employed, where the nonlinear iterative technique is known to perform better in comparison to linear techniques (noniterative) with a caveat that nonlinear techniques are computationally complex. Assuming that there is a linear dependency of solution between successive frames resulted in a linear inverse problem. This new framework with the combination of l(1)-norm based regularization can provide better robustness to noise and provide better contrast recovery compared to conventional l(2)-based techniques. Moreover, it is shown that the proposed l(1)-based technique is computationally efficient compared to its counterpart (l(2)-based one). The proposed framework requires a reasonably close estimate of the actual solution for the initial frame, and any suboptimal estimate leads to erroneous reconstruction results for the subsequent frames.

Nanomechanical and optical properties of highly a-axis oriented AlN films

This paper reports optical and nanomechanical properties of predominantly a-axis oriented AlN thin films. These films were deposited by reactive DC magnetron sputtering technique at an optimal target to substrate distance of 180 mm. X-ray rocking curve (FWHM = 52 arcsec) studies confirmed the preferred orientation. Spectroscopic ellipsometry revealed a refractive index of 1.93 at a wavelength of 546 nm. The hardness and elastic modulus of these films were 17 and 190 GPa, respectively, which are much higher than those reported earlier can be useful for piezoelectric films in bulk acoustic wave resonators. (C) 2012 American Institute of Physics. http://dx.doi.org/10.1063/1.4772204]

Nonquadratic penalization improves near-infrared diffuse optical tomography

A new approach that can easily incorporate any generic penalty function into the diffuse optical tomographic image reconstruction is introduced to show the utility of nonquadratic penalty functions. The penalty functions that were used include quadratic (l(2)), absolute (l(1)), Cauchy, and Geman-McClure. The regularization parameter in each of these cases was obtained automatically by using the generalized cross-validation method. The reconstruction results were systematically compared with each other via utilization of quantitative metrics, such as relative error and Pearson correlation. The reconstruction results indicate that, while the quadratic penalty may be able to provide better separation between two closely spaced targets, its contrast recovery capability is limited, and the sparseness promoting penalties, such as l(1), Cauchy, and Geman-McClure have better utility in reconstructing high-contrast and complex-shaped targets, with the Geman-McClure penalty being the most optimal one. (C) 2013 Optical Society of America

A two-axis in-plane motion measurement system based on optical beam deflection

Measurement of in-plane motion with high resolution and large bandwidth enables model-identification and real-time control of motion-stages. This paper presents an optical beam deflection based system for measurement of in-plane motion of both macro- and micro-scale motion stages. A curved reflector is integrated with the motion stage to achieve sensitivity to in-plane translational motion along two axes. Under optimal settings, the measurement system is shown to theoretically achieve sub-angstrom measurement resolution over a bandwidth in excess of 1 kHz and negligible cross-sensitivity to linear motion. Subsequently, the proposed technique is experimentally demonstrated by measuring the in-plane motion of a piezo flexure stage and a scanning probe microcantilever. For the former case, reflective spherical balls of different radii are employed to measure the in-plane motion and the measured sensitivities are shown to agree with theoretical values, on average, to within 8.3%. For the latter case, a prototype polydimethylsiloxane micro-reflector is integrated with the microcantilever. The measured in-plane motion of the microcantilever probe is used to identify nonlinearities and the transient dynamics of the piezo-stage upon which the probe is mounted. These are subsequently compensated by means of feedback control. (C) 2013 AIP Publishing LLC.

Going beyond Red with a Tri- and Tetracoordinate Boron Conjugate: Intriguing Near-IR Optical Properties and Applications in Anion Sensing

The design and synthesis of a new tri- and tetracoordinate boron conjugate is reported. The conjugate shows broad near-IR emission (similar to 625-850 nm) and is found to be a selective colorimetric and ratiometric sensor for fluoride ions.

Structural and optical investigation of semiconductor CdSe/CdS core-shell quantum dot thin films

Highly luminescent CdSe/CdS core-shell nanocrystals have been assembled on indium tin oxide (ITO) coated glass substrates using a wet synthesis route. The physical properties of the quantum dots (QD) have been investigated using X-ray diffraction, transmission electron microscopy and optical absorption spectroscopy techniques. These quantum dots showed a strong enhancement in the near band edge absorption. The in situ luminescence behavior has been interpreted in the light of the quantum confinement effect and induced strain in the core-shell structure.