Rapid casting of patterned vascular networks for perfusable engineered three-dimensional tissues

In the absence of perfusable vascular networks, three-dimensional (3D) engineered tissues densely populated with cells quickly develop a necrotic core. Yet the lack of a general approach to rapidly construct such networks remains a major challenge for 3D tissue culture. Here, we printed rigid 3D filament networks of carbohydrate glass, and used them as a cytocompatible sacrificial template in engineered tissues containing living cells to generate cylindrical networks that could be lined with endothelial cells and perfused with blood under high-pressure pulsatile flow. Because this simple vascular casting approach allows independent control of network geometry, endothelialization and extravascular tissue, it is compatible with a wide variety of cell types, synthetic and natural extracellular matrices, and crosslinking strategies. We also demonstrated that the perfused vascular channels sustained the metabolic function of primary rat hepatocytes in engineered tissue constructs that otherwise exhibited suppressed function in their core.     

Modeling anisotropic light propagation in a realistic model of the human head

A Monte Carlo model capable of describing photon migration in arbitrary three-dimensional geometry with spatially varying optical properties and tissue anisotropy is presented. We use the model to explore the effects of anisotropy for optical measurements of the human head. An anisotropic diffusion equation that corresponds to our Monte Carlo model is derived, and a comparison between the Monte Carlo model and the diffusion equation solution with finite elements is given.     

Semi-three-dimensional algorithm for time-resolved diffuse optical tomography by use of the generalized pulse spectrum technique

Although a foil three-dimensional (3-D) reconstruction with both 3-D forward and inverse models provide, the optimal solution for diffuse optical tomography (DOT), because of the 3-D nature of photon diffusion in tissue, it is computationally costly for both memory requirement and execution time in a conventional computing environment. Thus in practice there is motivation to develop an image reconstruction algorithm with dimensional reduction based on some modeling approximations. Here we have implemented a semi-3-D modified generalized pulse spectrum technique for time-resolved DOT, where a two-dimensional (2-D) distribution of optical properties is approximately assumed, while we retain 3-D distribution of photon migration in tissue. We have validated the proposed algorithm by reconstructing 3-D structural test objects from both numerically simulated and experimental date. We demonstrate our algorithm by comparing it with the calibrated 2-D reconstruction that is in widespread use as a shortcut to 3-D imaging and proving that the semi-3-D algorithm outperforms the calibrated 2-D algorithm.     

An automated approach for three-dimensional quantification of fibrillar structures in optically cleared soft biological tissues

We present a novel approach allowing for a simple, fast and automated morphological analysis of three-dimensional image stacks (z-stacks) featuring fibrillar structures from optically cleared soft biological tissues. Five non-atherosclerotic tissue samples from human abdominal aortas were used to outline the multi-purpose methodology, applicable to various tissue types. It yields a three-dimensional orientational distribution of relative amplitudes, representing the original collagen fibre morphology, identifies regions of isotropy where no preferred fibre orientations are observed and determines structural parameters throughout anisotropic regions for the analysis and numerical modelling of biomechanical quantities such as stress and strain. Our method combines optical tissue clearing with second-harmonic generation imaging, Fourier-based image analysis and maximum-likelihood estimation for distribution fitting. With a new sample preparation method for arteries, we present, for the first time to our knowledge, a continuous three-dimensional distribution of collagen fibres throughout the entire thickness of the aortic wall, revealing novel structural and organizational insights into the three arterial layers.     

Effects of zero-length and non-zero-length cross-linking reagents on the optical spectral properties and structures of collagen hydrogels

We compared the effects of zero-length cross-linkers 1-ethyl-3 (3dimethylaminopropyl) carbodiimide (EDC) and non-zero-length cross-linkers glycolaldehyde and glyceraldehyde on the optical and structural properties of three-dimensional (3D) collagen hydrogels. We evaluated these effects by multiphoton microscopy (MPM) that combined two-photon fluorescence (TPF) and second harmonic generation (SHG) contrasts and transmission electron microscopy (TEM). The collagen hydrogels were incubated separately with the above-mentioned reagents present at the concentration of 0.1 M. The incubation with glycolaldehyde and glyceraldehyde induced strong autofluorescence within the gels. We followed the formation of fluorescence with TPF signals in situ and in real time as well as characterized the micro- and nanostructures within cross-linked hydrogels by examining SHG and TEM images respectively. As detected in the SHG images, glycolaldehyde- and glyceraldehyde-modified 5-10 μm "fiberlike" collagen structures to longer, 20 μm and more, aggregated strands while EDC had minimal effect on the microstructure. TEM revealed that glycolaldehyde and glyceraldehyde either completely eliminated collagen's characteristic native fibrillar striations or generated uncharacteristic fibrils with extensions. EDC preserved the native striation patterns, decreased the fibril diameters and effectively homogenized the fibrils within hydrogels assembled at 1.8-4.68 g/L collagen concentrations and 37 °C. Our findings provide a clear understanding on how different cross-linking reagents have very different effects on the collagen hydrogels. This understanding is critical for advancing tissue engineering and wound healing applications.     

Determination of Teflon thickness with laser speckle. I. Potential for burn depth diagnosis

A quantitative method for determining the depth of burn eschar would aid surgeons in determining whether to excise and subsequently graft a burn wound. We hypothesize that tissue viability could be assessed by an analysis of the spatial modulation of near-field laser speckle by flowing blood. A feasibility study of the technique was performed with two-layer tissue phantoms used to simulate a burn wound. A sheet of polytetrafluoroethylene (PTFE) was used to simulate nonperfused burn eschar, and tissue perfusion within deeper layers was represented by Brownian motion from a scattering solution. A low-power He-Ne laser was focused onto the target, and the resulting speckle image was captured with a CCD camera and stored on a computer for further processing. The diameter of the speckle pattern was found to be directly proportional to the thickness of the overlying layer. These data suggest that the thickness of PTFE can be determined to ±100-μm accuracy with 95% confidence and may be suitable for burn depth detection in vivo.     

Volume image reconstruction for diffuse optical tomography

Optical tomography is a potential diagnostic method for visualizing optical properties of tissues in vivo. We present an optical tomography method that has been designed for imaging of the human testes, particularly for spectroscopic tumor differentiation. In this application we need to compute three-dimensional distributions of the optical contrast (absorption coefficient) in the tissue in real time. Thus we have given special care to elaborate an efficient inverse algorithm that takes the limitations of spatial resolution and data space point density into account. Our inverse solution is based on a linearization approach and a dedicated object space discretization. Furthermore, we introduce the concept of fuzzy voxels, which enables a reconstruction-inherent image smoothing.     

Determination of optical probe interrogation field of near-infrared reflectance: phantom and Monte Carlo study

An optical probe used to localize human brain tissues in vivo has been reported previously. It was able to sense the underlying tissue structure with an optical interrogation field, termed as "look ahead distance" (LAD). A new side-firing probe has been designed with its optical window along its side. We have defined the optical interrogation field of the new side probe as "look aside distance" (LASD). The purpose of this study is to understand the dependence of the LAD and LASD on the optical properties of tissue, the light source intensity, and the integration time of the detector, using experimental and computational methods. The results show that a decrease in light intensity does decrease the LAD and LASD and that an increase in integration time of detection may not necessarily improve the depths of LAD and LASD. Furthermore, Monte Carlo simulation results suggest that the LAD/LASD decreases with an increase in reduced scattering coefficient to a point, after which the LAD/LASD remains constant. We expect that an optical interrogation field of a tip or side probe is approximately 1-2 mm in white matter and 2-3.5 mm in gray matter. These conclusions will help us optimally manipulate the parameter settings during surgery and determine the spatial resolution of the probe.     

Robust inference of baseline optical properties of the human head with three-dimensional segmentation from magnetic resonance imaging

We model the capability of a small (6-optode) time-resolved diffuse optical tomography (DOT) system to infer baseline absorption and reduced scattering coefficients of the tissues of the human head (scalp, skull, and brain). Our heterogeneous three-dimensional diffusion forward model uses tissue geometry from segmented magnetic resonance (MR) data. Handling the inverse problem by use of Bayesian inference and introducing a realistic noise model, we predict coefficient error bars in terms of detected photon number and assumed model error. We demonstrate the large improvement that a MR-segmented model can provide: 2-10% error in brain coefficients (for 2 x 10(6) photons, 5% model error). We sample from the exact posterior and show robustness to numerical model error. This opens up the possibility of simultaneous DOT and MR for quantitative cortically constrained functional neuroimaging.     

Three-dimensional chiral plasmonic oligomers

The living world is chiral. Chirality or the handedness of a structure or molecule is at the heart of life itself. Recently, it has been shown that plasmonic structures exhibit unprecedented and gigantic chiral optical responses. Here we show that truly three-dimensional arrangements of plasmonic "meta-atoms" only exhibit a chiral optical response if similar plasmonic "atoms" are arranged in a handed fashion as we require resonant plasmonic coupling. Moreover, we demonstrate that such particle groupings, similarly to molecular systems, possess the capability to encode their three-dimensional arrangement in unique and well-modulated spectra making them ideal candidates for a three-dimensional chiral plasmon ruler. Our results are crucial for the future design and improvement of plasmonic chiral optical systems, for example, for ultrasensitive enantiomer sensing on the single molecule level.     

Three-dimensional positioning of optically trapped nanoparticles

We firstly demonstrate the three-dimensional (3D) measurement of a nanometer-sized sphere held in optical tweezers in water using an in-line digital holographic microscope with a green light emitting diode. Suppressing the movement with optical tweezers enabled us to detect the three-dimensional position of a polystyrene sphere with a diameter of 200 nm. The positioning resolutions of the microscope were 3.2 nm in the transverse direction and 3.4 nm in the axial direction, from the standard deviation of measurements of the 200 nm sphere fixed on glass. Changes in the Brownian motion in response to a change in the trapping laser power were measured. We also demonstrated that this holographic measurement is an effective method for determining the threshold power of the optical trapping.     

Signal-to-noise ratio study of full-field fourier-domain optical coherence tomography

We report a new approach in optical coherence tomography (OCT) called full-field Fourier-domain OCT (3F-OCT). A three-dimensional image of a sample is obtained by digital reconstruction of a three-dimensional data cube, acquired with a Fourier holography recording system, illuminated with a swept source. We present a theoretical and experimental study of the signal-to-noise ratio of the 3F-OCT approach versus serial image acquisition (flying-spot OCT) approach.     

Direct patterning of three-dimensional periodic nanostructures by surface-plasmon-assisted nanolithography

The technical applications of three-dimensional (3D) nanostructures demand a fabrication technique that is convenient and yet offers design flexibility. We describe here a nanofabrication technique called surface-plasmon-assisted three-dimensional nanolithography (3D-SPAN). By utilizing optical near-field interference patterns generated by surface plasmons (SPs), we have fabricated different 2D/3D periodic polymeric nanostructures in a typical photolithography setup. We have also shown here that the nanostructures fabricated by 3D-SPAN can be controlled easily in terms of size, layout, and defects by designing the SPAN mask. Because of its design flexibility and fabrication convenience, 3D-SPAN could be used to develop "photonics on a chip", where signal processing is carried out by photons instead of electrons and be extended to other applications of direct-writing 3D functional nanostructures.     

Equivalent-optical-waveguide model for the analysis of optical waveguides by means of an asymptotic effective-index method

We present a theoretical method that makes it possible to analyze three-dimensional (3-D) integrated optical waveguides with arbitrary refractive-index profiles. With this method it is easy to obtain effective indexes, propagation constants, and coupling-switching properties of planar and channel optical waveguides. This theoretical approach involves one's modeling the original optical waveguide by means of an equivalent optical waveguide whose effective index is evaluated by the application of a technique that we call the asymptotic effective-index method. The numerical values show good convergence and accuracy for effective indexes, propagation constants, and coupling-switching characteristics. Theoretical and experimental results are given.     

Application of plasmonic bowtie nanoantenna arrays for optical trapping, stacking, and sorting

We present the use of Au bowtie nanoantenna arrays (BNAs) for highly efficient, multipurpose particle manipulation with unprecedented low input power and low-numerical aperture (NA) focusing. Optical trapping efficiencies measured are up to 20× the efficiencies of conventional high-NA optical traps and are among the highest reported to date. Empirically obtained plasmonic optical trapping "phase diagrams" are introduced to detail the trapping response of the BNAs as a function of input power, wavelength, polarization, particle diameter, and BNA array spacing (number density). Using these diagrams, parameters are chosen, employing strictly the degrees-of-freedom of the input light, to engineer specific trapping tasks including (1) dexterous, single-particle trapping and manipulation, (2) trapping and manipulation of two- and three-dimensional particle clusters, and (3) particle sorting. The use of low input power densities (power and NA) suggests that this bowtie nanoantenna trapping system will be particularly attractive for lab-on-a-chip technology or biological applications aimed at reducing specimen photodamage.     

Two-dimensional correlation analysis study of the photo-induced molecular reorientations of photosensitive polyester film containing 1,4-phenylenediacryloyl units in the backbone

The photochemical reaction and molecular reorientation of a novel photosensitive polyester, poly[oxy(4-n-butyl-3,5-benzoate)oxy-1,4-phenylenediacryloyl] (PPDA-C4BZ), which contains n-butyl side groups and 1,4-phenylenediacryloyl units (PDA chromophores) in the main chain, are reported in detail. We applied two-dimensional (2D) correlation analysis for the infrared (IR) and ultraviolet (UV) absorption spectra of nanoscaled films of PPDA-C4BZ to establish the sequence of the photo-induced segmental reorientations that result from UV irradiation. The photochemical reaction was found to have a greater effect on the polymer's main chains than on its side groups and to induce the reorientation of the polymer molecules. In particular, a cycloaddition process occurs first in the PDA chromophore units and then the local reorientation of the polymer molecules is induced. Namely, such photodimerization of the PDA chromophores induces the molecular reorientations of the PDA chromophores and the benzoate units in the main chain. The photo-induced molecular reorientations occur in the following sequence: photodimerization --> benzoate units --> PDA chromophores --> n-butyl side groups. In addition, a two-dimensional map of the first derivatives of the UV absorption spectra with respect to the exposure energy provided evidence of the formation of head-to-head aggregates (i.e., H-aggregates) of PPDA-C4BZ molecules.     

Reflectivity of the gyroid biophotonic crystals in the ventral wing scales of the Green Hairstreak butterfly,Callophrys rubi

We present a comparison of the computer simulation data of gyroid nanostructures with optical measurements (reflectivity spectra and scattering diagrams) of ventral wing scales of the Green Hairstreak butterfly, Callophrys rubi. We demonstrate that the omnidirectional green colour arises from the gyroid cuticular structure grown in the domains of different orientation. We also show that this three-dimensional structure, operating as a biophotonic crystal, gives rise to various polarization effects. We briefly discuss the possible biological utility of the green coloration and polarization effects.     

On the anharmonic contribution to the specific heat of monatomic face-centered cubic crystals

We have presented a method for an exact calculation of the two lowest-order, cubic (F ₃) and quartic (F ₄), perturbation terms in the Helmholtz free energy (F) of an anharmonic crystal in the high temperature limit. The method is applicable to a nearest-neighbor central force model of a monatomic fcc crystal for any phenomenological two-body potential (r). The calculation of F ₃ and F ₄ requires a knowledge of the six dimensionless Brillouin zone (BZ) sums, as a function of a parameter a ₁ depending on the first and second derivatives of (r). These sums are calculated to a high degree of accuracy for a mesh size of 308,000 points in the whole BZ in the range –0.1a ₁+0.1 in steps of 0.02. The linear temperature dependent anharmonic contribution to the specific heat at constant volume, calculated for the elements Pb, Ag, Ni, Cu, Al, Ca, and Sr, from the Morse and Rydberg potentials, is found to be positive in all cases, with the exception of Pb. In this case the Morse potential gives a negative sign. The predictions of theory are in agreement with experiments where the data is available (e.g., Cu, Al, and Pb).     

Noncontact optical imaging in mice with full angular coverage and automatic surface extraction

During the past decade, optical imaging combined with tomographic approaches has proved its potential in offering quantitative three-dimensional spatial maps of chromophore or fluorophore concentration in vivo. Due to its direct application in biology and biomedicine, diffuse optical tomography (DOT) and its fluorescence counterpart, fluorescence molecular tomography (FMT), have benefited from an increase in devoted research and new experimental and theoretical developments, giving rise to a new imaging modality. The most recent advances in FMT and DOT are based on the capability of collecting large data sets by using CCDs as detectors, and on the ability to include multiple projections through recently developed noncontact approaches. For these to be implemented, we have developed an imaging setup that enables three-dimensional imaging of arbitrary shapes in fluorescence or absorption mode that is appropriate for small animal imaging. This is achieved by implementing a noncontact approach both for sources and detectors and coregistering surface geometry measurements using the same CCD camera. A thresholded shadowgrammetry approach is applied to the geometry measurements to retrieve the surface mesh. We present the evaluation of the system and method in recovering three-dimensional surfaces from phantom data and live mice. The approach is used to map the measured in vivo fluorescence data onto the tissue surface by making use of the free-space propagation equations, as well as to reconstruct fluorescence concentrations inside highly scattering tissuelike phantom samples. Finally, the potential use of this setup for in vivo small animal imaging and its impact on biomedical research is discussed.     

Analysis of optical waveguide structures by use of a combined finite-difference/finite-difference time-domain method

We present a method for full-wave characterization of optical waveguide structures. The method computes mode-propagation constants and cross-sectional field profiles from a straight forward discretization of Maxwell's equations. These modes are directly excited in a three-dimensional finite-difference time-domain simulation to obtain optical field transmission and reflection coefficients for arbitrary waveguide discontinuities. The implementation uses the perfectly-matched-layer technique to absorb both guided modes and radiated fields. A scattered-field formulation is also utilized to allow accurate determination of weak scattered-field strengths. Individual three-dimensional waveguide sections are cascaded by S-parameter analysis. A complete 10(4)-section Bragg resonator is successfully simulated with the method.