Faculty Bibliography

Red blood cells (RBCs) transport oxygen to tissues and remove carbon dioxide. Diffuse optical flowmetry (DOF) assesses deep tissue RBC dynamics by measuring coherent fluctuations of multiply scattered near-infrared light intensity. While classical DOF measurements empirically correlate with blood flow, they remain far-removed from light scattering physics and difficult to interpret in layered media. To advance DOF measurements closer to the physics, here we introduce an interferometric technique, surmounting challenges of bulk motion to apply it in awake humans. We reveal two measurement dimensions: optical phase, and time-of-flight (TOF), the latter with 22 picosecond resolution. With this multidimensional data, we directly confirm the unordered, or Brownian, nature of optically probed RBC dynamics typically assumed in classical DOF. We illustrate how incorrect absorption assumptions, anisotropic RBC scattering, and layered tissues may confound classical DOF. By comparison, our direct method enables accurate and comprehensive assessment of blood flow dynamics in humans.

Across optics and photonics, excess intensity noise is often considered a liability. Here, we show that excess noise in broadband supercontinuum and superluminescent diode light sources encodes each spectral channel with unique intensity fluctuations, which actually serve a useful purpose. Specifically, we report that excess noise correlations can both characterize the spectral resolution of spectrometers and enable cross-calibration of their wavelengths across a broad bandwidth. Relative to previous methods that use broadband interferometry and narrow linewidth lasers to characterize and calibrate spectrometers, our approach is simple, comprehensive, and rapid enough to be deployed during spectrometer alignment. First, we employ this approach to aid alignment and reduce the depth-dependent degradation of the sensitivity and axial resolution in a spectrometer-based optical coherence tomography (OCT) system, revealing a new outer retinal band. Second, we achieve a pixel-to-pixel correspondence between two otherwise disparate spectrometers, enabling a robust comparison of their respective measurements. Thus, excess intensity noise has useful applications in optics and photonics.

Studies of flow-metabolism coupling often presume that microvessel architecture is a surrogate for blood flow. To test this assumption, we introduce an in vivo Dynamic Contrast Optical Coherence Tomography (DyC-OCT) method to quantify layer-resolved microvascular blood flow and volume across the full depth of the mouse neocortex, where the angioarchitecture has been previously described. First, we cross-validate average DyC-OCT cortical flow against conventional Doppler OCT flow. Next, with laminar DyC-OCT, we discover that layer 4 consistently exhibits the highest microvascular blood flow, approximately two-fold higher than the outer cortical layers. While flow differences between layers are well-explained by microvascular volume and density, flow differences between subjects are better explained by transit time. Finally, from layer-resolved tracer enhancement, we also infer that microvascular hematocrit increases in deep cortical layers, consistent with predictions of plasma skimming. Altogether, our results show that while the cortical blood supply derives mainly from the pial surface, laminar hemodynamics ensure that the energetic needs of individual cortical layers are met. The laminar trends reported here provide data that links predictions based on the cortical angioarchitecture to cerebrovascular physiology in vivo.

In biological tissue, longer near-infrared wavelengths generally experience less scattering and more water absorption. Here we demonstrate an optical coherence tomography (OCT) system centered at 2.1 microns, whose bandwidth falls in the 2.2 micron water absorption optical window, for in vivo imaging of the rodent brain. We show in vivo that at 2.1 microns, the OCT signal is actually attenuated less in cranial bone than at 1.3 microns, and is also less susceptible to multiple scattering tails. We also show that the 2.2 micron window enables direct spectroscopic OCT assessment of tissue water content. We conclude that with further optimization, 2.2 micron OCT will have advantages in low-water-content tissue such as bone, as well as applications where extensive averaging is possible to compensate absorption losses.

Visible light optical coherence tomography (OCT) theoretically provides finer axial resolution than near-infrared OCT for a given wavelength bandwidth. To realize this potential in the human retina in vivo, the unique technical challenges of visible light OCT must be addressed. We introduce three advances to further the performance of visible light OCT in the human retina. First, we incorporate a grating light valve spatial light modulator (GLV-SLM) spectral shaping stage to modify the source spectrum. This enables comfortable subject alignment with a red light spectrum, and image acquisition with a broad "white light" spectrum, shaped to minimize sidelobes. Second, we develop a novel, Fourier transform-free, software axial motion tracking algorithm with fast, magnetically actuated stage to maintain near-optimal axial resolution and sensitivity in the presence of eye motion. Third, we implement spatially dependent numerical dispersion compensation for the first time in the human eye in vivo. In vivo human retinal OCT images clearly show that the inner plexiform layer consists of 3 hyper-reflective bands and 2 hypo-reflective bands, corresponding with the standard anatomical division of the IPL. Wavelength-dependent images of the outer retina suggest that, beyond merely improving the axial resolution, shorter wavelength visible light may also provide unique advantages for visualizing Bruch's membrane.

Visible light optical coherence tomography (OCT) has recently emerged in retinal imaging, with claims of micrometer-scale axial resolution and multi-color (sub-band) imaging. Here, we show that the large dispersion of optical glass and aqueous media, together with broad optical bandwidths often used in visible light OCT, compromises both of these claims. To rectify this, we introduce the notion of spatially dependent (i.e., depth and transverse position-dependent) dispersion. We use a novel sub-band, sub-image correlation algorithm to estimate spatially dependent dispersion in our 109 nm bandwidth visible light OCT mouse retinal imaging system centered at 587 nm. After carefully compensating spatially dependent dispersion, we achieve delineation of fine outer retinal bands in mouse strains of varying pigmentation. Spatially dependent dispersion correction is critical for broader bandwidths and shorter visible wavelengths.

Quantifying light transport in turbid media is a long-standing challenge. This challenge arises from the difficulty in experimentally separating unscattered, ballistic light from forward scattered light. Correlation gating is a new approach that numerically separates light paths based on statistical dynamics of the optical field. Here we apply correlation gating with interferometric near-infrared spectroscopy (iNIRS) to separate and independently quantify ballistic and scattered light transmitted through thick samples. First, we present evidence that correlation gating improves the isolation of ballistic light in a thick, intrinsically dynamic medium with Brownian motion. Then, from a single set of iNIRS transmission measurements, we determine the ballistic attenuation coefficient and group refractive index from the time-of-flight (TOF) resolved static intensity, and we determine the reduced scattering and absorption coefficients from the diffusive part of the TOF resolved dynamic intensity. Finally, we show that correlation gating is applicable in intrinsically static media in which motion is induced externally. Thus, for the first time, to the best of our knowledge, the key optical properties of a turbid medium can be derived from a single set of transmission measurements.

Chromatic aberrations are an important design consideration in high resolution, high bandwidth, refractive imaging systems that use visible light. Here, we present a fiber-based spectral/Fourier domain, visible light OCT ophthalmoscope corrected for the average longitudinal chromatic aberration (LCA) of the human eye. Analysis of complex speckles from in vivo retinal images showed that achromatization resulted in a speckle autocorrelation function that was ~20% narrower in the axial direction, but unchanged in the transverse direction. In images from the improved, achromatized system, the separation between Bruch's membrane (BM), the retinal pigment epithelium (RPE), and the outer segment tips clearly emerged across the entire 6.5 mm field-of-view, enabling segmentation and morphometry of BM and the RPE in a human subject. Finally, cross-sectional images depicted distinct inner retinal layers with high resolution. Thus, with chromatic aberration compensation, visible light OCT can achieve volume resolutions and retinal image quality that matches or exceeds ultrahigh resolution near-infrared OCT systems with no monochromatic aberration compensation.

We introduce the feature issue on the Optics in the Life Sciences Congress held on April 2-5, 2017 in San Diego, CA. The Congress consisted of 5 topical symposia: (i) Bio-optics Design and Application; (ii) Novel Techniques in Microscopy; (iii) Optical Molecular Probes, Imaging and Drug Delivery; (iv) Optical Trapping Applications; and (v) Optics and the Brain. These separate symposia also held joint sessions of common interest. The following highlights some of the topics from the Congress.