Contrast and resolution analysis of iterative angular domain optical projection tomography.

In Angular Domain Imaging, image contrast and resolution are position dependent. The objective of this work was to characterize the contrast and resolution of an ADI system at a multitude of locations within the imaging plane, then compare the reconstructions of different targets using filtered back projection and iterative reconstruction algorithms. Contrast varied significantly with depth and minimally with lateral position, while resolution varied significantly with lateral position and minimally with depth. The iterative reconstruction algorithm was robust against ring and streak artifacts. The back projection reconstructions suffered from artifacts related to a lack of projection data.

Transmission and fluorescence angular domain optical projection tomography of turbid media.

When imaging through turbid media, objects are often blurred by scattered light. An optical collimator (i.e., an angular filter array) improves images by accepting only photons propagating within a narrow solid angle about the direction of the incident light. These photons are expected to participate in a limited number of small-angle scattering events, maintaining their original propagation direction and, finally, contributing to the development of a faithful image of an object within a turbid medium. The collimation method, also referred to as angular domain imaging (ADI), applies to a see-through configuration where the incident collimated light beam can be aligned with the collimator in a transillumination mode of operation. In this paper, we present angular domain optical projection tomography (ADOPT), a method that can extract depth information of optical contrast in turbid media with high longitudinal resolution based on ADI technology. The resolution of the ADI system has been tested over various depths in a 5 cm optical cuvette using a resolution target suspended in a homogeneous turbid medium. The ADOPT system reconstructed images from a series of angular domain projections collected at angular intervals. The system was used to measure the attenuation of an absorbing target in transmission mode (t-ADOPT) and to measure the light emitting from a fluorescent target (f-ADOPT). Tissue-mimicking phantoms were used to validate the performance of the method. In the t-ADOPT configuration, a background scattered light estimation and subtraction methodology was introduced to improve the imaging contrast. A target consisting of two graphite rods (0.9 mm diameter) was suspended in the cuvette by a rotation stage. An Indocyanine Green-filled glass rod was used as an imaging target in the f-ADOPT arrangement. The target was placed in a manner such that the line of laser light was perpendicular to the longitudinal axis of the rods. Several projections were collected at increments of 1.8 degrees and compiled into a sinogram. A transverse image was reconstructed from the sinogram by using filtered backprojection and image contrast was improved by experimental scatter measurements using a wedge prism and an image processing algorithm. The submillimeter target embedded in a 2 cm thick scattering medium (reduced scattering coefficient < or = 2.4 cm(-1)) was discernable in both the sinograms and the reconstructed images. In the f-ADOPT system, fluorescent line targets <1 cm in diameter embedded in a 2 cm thick scattering medium (reduced scattering coefficient < or = 0.8 cm(-1)) were discernable in both the sinograms and the reconstructed images. The proposed method could be used as the basis to construct an optical tomographic scanner for simultaneous absorption and fluorescence-based imaging of biological specimens (i.e., up to 7 mm across).

Successive order, multiple scattering of two-term Henyey-Greenstein phase functions.

An analytic solution to the problem of determining photon direction after successive scatterings in an infinite, homogeneous, isotropic medium, where each scattering event is in accordance with a two-term Henyey-Greenstein phase function, is presented and compared against Monte Carlo simulation results. The photon direction is described by a probability density function of the dot product of the initial direction and the direction after multiple scattering events, and it is found that such a probability density function can be represented as a weighted series of one-term Henyey-Greenstein phase functions.

Image contrast enhancement in angular domain optical imaging of turbid media.

Imaging structures within a turbid medium using Angular Domain Imaging (ADI) employs an angular filter array to separate weakly scattered photons from those that are highly scattered. At high scattering coefficients, ADI contrast declines due to the large fraction of non-uniform background scattered light still within the acceptance angle. This paper demonstrates various methods to enhance the image contrast in ADI. Experiments where a wedge prism was used to deviate the laser source so that scattered photons could be imaged and subtracted from the image obtained by standard ADI provided the greatest improvement in image contrast.

Multi-spectral angular domain optical imaging in biological tissues using diode laser sources.

Angular Domain Imaging (ADI) employs micromachined angular filter to detect non-scattered photons that pass through the micro-scale tunnels unattenuated while scattered photons are rejected. This paper describes the construction of an ADI system utilizing diode lasers at three different wavelengths in the range of the red and near infrared spectrum. Experiments are performed to verify the feasibility of ADI at multi-wavelengths. ADI results of chicken breast as a biological scattering medium are presented for different thicknesses. A spatial resolution of <0.5 mm is achieved with 5 mm thick chicken breast using a 975 nm diode laser source.

Effects of chemical and physical parameters in the generation of microspheres by hydrodynamic flow focusing.

Hydrodynamic flow focusing is a seminal, easy-to-use technology for micro- and nanodroplet generation. It is characterized by the co-axial focusing of two (or more) immiscible liquid streams forced through a small orifice. In this method, the outer continuous phase has a much higher flow velocity than the inner disperse phase. While passing through the orifice, the prevailing pressure drop and shear stress force the inner phase to break up into uniform droplets. Using a biodegradable poly(lactide-co-glycolide) (PLGA) polymer solution as the disperse phase, monodisperse and user-defined polymer micro- and nanospheres can be generated. Here we present a consecutive parameter study of hydrodynamic flow focusing to study the effect of chemical and physical parameters that effect the dispersity of the droplets generated in the 1-5 µm range. The parameter study shows the applicability and challenges of hydrodynamic flow focusing in the preparation of biodegradable microspheres. Applications for microspheres made with this method can be found in the medical, pharmaceutical and technical fields.

Angular domain transillumination imaging optimization with an ultrafast gated camera.

By employing high-aspect-ratio parallel microchannels as an angular filter, quasiballistic photons sensitive to internal structures in a turbid medium can be captured. Scattered photons exiting the turbid medium typically exhibit trajectories with random angles compared to the initial trajectory and are mostly rejected by the filter. However, angular filter arrays cannot differentiate between quasiballistic photons (early arriving) and photons that happen to attain a scattered trajectory that is within the acceptance angle (late arriving). Therefore, we have two objectives: (1) to experimentally characterize the angular distribution and proportion of minimally deviated quasiballistic photons and multiply scattered photons in a turbid medium and (2) to combine time and angular gating principles so that early and late arriving photons can be distinguished. From the angular distribution data, the angular filter with angular acceptance about 0.4 deg yields the highest image contrast for transillumination images. The use of angular domain imaging(ADI) with time-gating enables visualization of submillimeter absorbing objects with approximately seven times higher image contrast compared to ADI in a turbid medium with a scattering level of six times the reduced mean free path.

Angular domain fluorescence imaging for small animal research.

We describe a novel macroscopic fluorescent imaging technique called angular domain fluorescence imaging (ADFI) applicable to the detection of fluorophores embedded in biological tissues. The method exploits the collimation detection capabilities of an angular filter array (AFA). The AFA uses the principle of acceptance angle filtration to extract minimally scattered photons emitted from fluorophores deep within tissue. Our goal was to develop an ADFI system for imaging near-infrared fluorescent markers for small animal imaging. According to the experimental results, the ADFI system offered higher resolution and contrast compared to a conventional lens and lens-pinhole fluorescent detection system. Furthermore, ADFI of a hairless mouse injected with a fluorescent bone marker revealed vertebral structural and morphometric data that correlated well with data derived from volumetric x-ray computed tomography images. The results suggested that ADFI is a useful technique for submillimeter mapping of the distribution of fluorescent biomarkers in small animals.

Use of hydrodynamic flow focusing for the generation of biodegradable camptothecin-loaded polymer microspheres.

The present study was conducted to investigate the use of hydrodynamic flow focusing for the generation of biodegradable polymer microspheres encapsulating the anticancer drug camptothecin. Poly(D,L-lactide-co-glycolide) (PLGA) and poly(L-lactide) (PLA) were used as the matrix materials. Camptothecin was dissolved in the disperse phase and microspheres with a mean size between 2 and 3 microm generated using hydrodynamic flow focusing. When up to 1 wt.% of the drug was added to PLA, the drug encapsulation efficiency was 64%. For PLGA, the drug encapsulation efficiency was between 39 and 46%. Drug release from PLA particles was rapid and complete within 6 h, while drug release from PLGA particles showed no burst effect and followed a first order release profile. The encapsulated camptothecin stayed in its active lactone form, as shown by HPLC, and was able to exert cell toxic effects as shown by a cell viability assay. Hydrodynamic flow focusing is a promising tool for the preparation of drug-releasing biodegradable microspheres typically made by solvent evaporation and/or solvent extraction, as indicated by the successful encapsulation of the anticancer drug camptothecin.