Jordan Dwelle

Thickness, Phase Retardation, Birefringence, and Reflectance of the Retinal Nerve Fiber Layer in Normal and Glaucomatous Non-Human Primates

PURPOSE We identified candidate optical coherence tomography (OCT) markers for early glaucoma diagnosis. Time variation of retinal nerve fiber layer (RNFL) thickness, phase retardation, birefringence, and reflectance using polarization sensitive optical coherence tomography (PS-OCT) were measured in three non-human primates with induced glaucoma in one eye. We characterized time variation of RNFL thickness, phase retardation, birefringence, and reflectance with elevated intraocular pressure (IOP). METHODS One eye of each of three non-human primates was laser treated to increase IOP. Each primate was followed for a 30-week period. PS-OCT measurements were recorded at weekly intervals. Reflectance index (RI) is introduced to characterize RNFL reflectance. Associations between elevated IOP and RNFL thickness, phase retardation, birefringence, and reflectance were characterized in seven regions (entire retina, inner and outer rings, and nasal, temporal, superior and inferior quadrants) by linear and non-linear mixed-effects models. RESULTS Elevated IOP was achieved in three non-human primate eyes with an average increase of 13 mm Hg over the study period. Elevated IOP was associated with decreased RNFL thickness in the nasal region (P = 0.0002), decreased RNFL phase retardation in the superior (P = 0.046) and inferior (P = 0.021) regions, decreased RNFL birefringence in the nasal (P = 0.002) and inferior (P = 0.029) regions, and loss of RNFL reflectance in the outer rings (P = 0.018). When averaged over the entire retinal area, only RNFL reflectance showed a significant decrease (P = 0.028). CONCLUSIONS Of the measured parameters, decreased RNFL reflectance was the most robust correlate with glaucomatous damage. Candidate cellular mechanisms are considered for decreased RNFL reflectance, including mitochondrial dysfunction and retinal ganglion cell apoptosis.

Birefringence measurement of the retinal nerve fiber layer by swept source polarization sensitive optical coherence tomography

A Swept Source Polarization-Sensitive Optical Coherence Tomography (SS-PS-OCT) instrument has been designed, constructed, and verified to provide high sensitivity depth-resolved birefringence and phase retardation measurements of the retinal nerve fiber layer. The swept-source laser had a center wavelength of 1059 nm, a full-width-half-max spectral bandwidth of 58 nm and an A-line scan rate of 34 KHz. Power incident on the cornea was 440 µW and measured axial resolution was 17 µm in air. A multiple polarization state nonlinear fitting algorithm was used to measure retinal birefringence with low uncertainty. Maps of RNFL phase retardation in a subject measured with SS-PS-OCT compare well with those generated using a commercial scanning laser polarimetry instrument. Peak-to-valley variation of RNFL birefringence given here is less than values previously reported at 840nm.

Combined two-photon luminescence microscopy and OCT for macrophage detection in the hypercholesterolemic rabbit aorta using plasmonic gold nanorose

The macrophage is an important early cellular marker related to risk of future rupture of atherosclerotic plaques. Two‐channel two‐photon luminescence (TPL) microscopy combined with optical coherence tomography (OCT) was used to detect, and further characterize the distribution of aorta‐based macrophages using plasmonic gold nanorose as an imaging contrast agent.

Path-length-multiplexed scattering-angle-diverse optical coherence tomography for retinal imaging.

A low-resolution path-length-multiplexed scattering angle diverse optical coherence tomography (PM-SAD-OCT) is constructed to investigate the scattering properties of the retinal nerve fiber layer (RNFL). Low-resolution PM-SAD-OCT retinal images acquired from a healthy human subject show the variation of RNFL scattering properties at retinal locations around the optic nerve head. The results are consistent with known retinal ganglion cell neural anatomy and principles of light scattering. Application of PM-SAD-OCT may provide potentially valuable diagnostic information for clinical retinal imaging.

Dual-wavelength multifrequency photothermal wave imaging combined with optical coherence tomography for macrophage and lipid detection in atherosclerotic plaques using gold nanoparticles

The objective of this study was to assess the ability of combined photothermal wave (PTW) imaging and optical coherence tomography (OCT) to detect, and further characterize the distribution of macrophages (having taken up plasmonic gold nanorose as a contrast agent) and lipid deposits in atherosclerotic plaques. Aortas with atherosclerotic plaques were harvested from nine male New Zealand white rabbits divided into nanorose- and saline-injected groups and were imaged by dual-wavelength (800 and 1210 nm) multifrequency (0.1, 1 and 4 Hz) PTW imaging in combination with OCT. Amplitude PTW images suggest that lateral and depth distribution of nanorose-loaded macrophages (confirmed by two-photon luminescence microscopy and RAM-11 macrophage stain) and lipid deposits can be identified at selected modulation frequencies. Radiometric temperature increase and modulation amplitude of superficial nanoroses in response to 4 Hz laser irradiation (800 nm) were significantly higher than native plaque (P<0.001). Amplitude PTW images (4 Hz) were merged into a coregistered OCT image, suggesting that superficial nanorose-loaded macrophages are distributed at shoulders on the upstream side of atherosclerotic plaques (P<0.001) at edges of lipid deposits. Results suggest that combined PTW-OCT imaging can simultaneously reveal plaque structure and composition, permitting characterization of nanorose-loaded macrophages and lipid deposits in atherosclerotic plaques.

Fourier optics analysis of phase-mask-based path-length-multiplexed optical coherence tomography

Optical coherence tomography (OCT) is an imaging technique that constructs a depth-resolved image by measuring the optical path-length difference between broadband light backscattered from a sample and a reference surface. For many OCT sample arm optical configurations, sample illumination and backscattered light detection share a common path. When a phase mask is placed in the sample path, features in the detected signal are observed, which suggests that an analysis of a generic common path OCT imaging system is warranted. In this study, we present a Fourier optics analysis using a Fresnel diffraction approximation of an OCT system with a path-length-multiplexing element (PME) inserted in the sample arm optics. The analysis may be generalized for most phase-mask-based OCT systems. A radial-angle-diverse PME is analyzed in detail, and the point spread function, coherent transfer function, sensitivity of backscattering angular diversity detection, and signal formation in terms of sample spatial frequency are simulated and discussed. The analysis reveals important imaging features and application limitations of OCT imaging systems with a phase mask in the sample path optics.

Differences in forward angular light scattering distributions between M1 and M2 macrophages.

Abstract. The ability to distinguish macrophage subtypes noninvasively could have diagnostic potential in cancer, atherosclerosis, and diabetes, where polarized M1 and M2 macrophages play critical and often opposing roles. Current methods to distinguish macrophage subtypes rely on tissue biopsy. Optical imaging techniques based on light scattering are of interest as they can be translated into biopsy-free strategies. Because mitochondria are relatively strong subcellular light scattering centers, and M2 macrophages are known to have enhanced mitochondrial biogenesis compared to M1, we hypothesized that M1 and M2 macrophages may have different angular light scattering profiles. To test this, we developed an in vitro angle-resolved forward light scattering measurement system. We found that M1 and M2 macrophage monolayers scatter relatively unequal amounts of light in the forward direction between 1.6 deg and 3.2 deg with M2 forward scattering significantly more light than M1 at increasing angles. The ratio of forward scattering can be used to identify the polarization state of macrophage populations in culture.

Noise model for polarization-sensitive Optical Coherence Tomography

Characterizing and quantifying noise sources in birefringence imaging with polarization-sensitive optical coherence tomography (PS-OCT) is necessary for the development of efficient noise reduction techniques for real-time clinical PS-OCT imaging. We propose three noise regimes based on the strength of specimen backscattering and dominated by different noise sources. We introduce a model that predicts noise effects in two regimes. The model includes source/detector intensity noise, and couples speckle effects with the longitudinal delays due to instrument and specimen birefringence to create realistic noise on simulated orthogonal interference fringe amplitudes and on their relative phases. Experimental examples of the three regimes are presented and in two of them, qualitative agreement between the model and experimental data is demonstrated.

Relationship between birefringence and neurotubule density in the primate retinal nerve fiber layer

The relationship between retinal nerve fiber layer (RNFL) birefringence (&Dgr;n) and neurotubule density (NTD, retinal ganglion cell (RGC) neurotubules per unit RNFL area) was investigated by correlating measurements of these two parameters in 1 eye of a healthy cynomolgus monkey. Phase retardation per unit depth (PR/UD, proportional to &Dgr;n) was measured at 5.6-15o intervals around the optic nerve head (ONH) with an enhanced polarization-sensitive optical coherence tomography (EPS-OCT) instrument. Transverse tissue sections containing 3 RGC nerve fiber bundles from each peripapillary RNFL octant were imaged with a transmission electron microscope (TEM). Morphological measurements taken in TEM images were used by a novel algorithm to estimate NTD. Registered PR/UD and NTD data were then correlated using single- and multi-level models, yielding correlation coefficients in the range 0.49 ⩽ r ⩽ 0.61 (0.06 ⩽ P ⩽ 0.11). It was found that in order for the single-level correlation coefficient (r = 0.61) to be statistically significant (P ⩽ 0.05) and powerful (Power ⩾ 80%), NTD measurements in at least 16, rather than 8, RNFL sectors were needed. Interestingly, a single-level correlation coefficient of r = 0.81 (P = 0.01) was calculated between octant-averaged PR/UD and RGC axoplasmic area (Ax, axon area less non-cytoskeletal organelle area) mode. Ax represents a RGC axons neurotubule-inhabitable area. Intuitively, a strong relationship should exist between Ax and neurotubule number if neurotubules provide the primary structural support for RGC axons and structural requirements are the same in all RGC axons. If this relationship exists, error resulting from NTD estimation methods or preservation artifacts may have caused lower observed correlations of PR/UD with NTD than with Ax mode, and more accurate methods of measuring in vivo NTD may be required to determine an accurate relationship between RNFL birefringence and NTD.