We present a novel optical coherence tomography (OCT) system design that employs coherence revival-based
heterodyning and polarization encoding to simultaneously image the ocular anterior segment and the retina. Coherence
revival heterodyning allows for multiple depths within a sample to be simultaneously imaged and frequency encoded by
carefully controlling the optical pathlength of each sample path. A polarization-encoded sample arm was used to direct
orthogonal polarizations to the anterior segment and retina. This design is a significant step toward realizing whole-eye
OCT, which would enable customized ray-traced modeling of patient eyes to improve refractive surgical interventions,
as well as the elimination of optical artifacts in retinal OCT diagnostics. We demonstrated the feasibility of this system
by acquiring images of the anterior segments and retinas of healthy human volunteers.
KEYWORDS: Spectroscopy, Optical coherence tomography, Doppler effect, Doppler tomography, Data acquisition, Signal to noise ratio, Velocity measurements, Signal attenuation, Imaging systems, Data centers
Recent advances in Doppler and variance techniques have enabled high sensitivity imaging in regions of
biological flow to measure blood velocities and vascular perfusion. In recent years, the sensitivity and imaging speed
benefits of Fourier domain OCT have become apparent. Spectrometer-based and wavelength-swept implementations
have both undergone rapid development. Comparative analysis of the potential benefits and limitations for the various
configurations would be useful for matching technology capabilities to specific clinical problems. Here we take a first
step in such a comparative analysis by presenting theoretical predictions and experimental results characterizing the
lower and upper observable velocity limits in spectrometer-based versus swept-source Doppler OCT. Furthermore, we
characterize the washout limit, the velocity at which signal degradation results in loss of flow information. We present
comparative results from phantom flow data as well as retinal data obtained with a commercial spectrometer OCT
system and a custom high-speed swept-source retinal OCT system.
Phase-sensitive adjuncts to optical coherence tomography (OCT) including Doppler and polarization-sensitive implementations allow for quantitative depth-resolved measurements of sample structure and dynamics including fluid flows and orientation of birefringent structures. The development of Fourier-domain OCT (FDOCT), particularly spectrometer-based spectral-domain systems with no moving parts (spectral-domain OCT or SDOCT), have greatly enhanced the phase stability of OCT systems particularly when implemented in a common-path geometry. The latter combination has given rise to a new class of nm-scale sensitive quantitative phase microscopies we have termed spectral domain phase microscopy. However, the phase information in all of these techniques suffers from a 2π ambiguity that limits resolvable pathlength differences to less than half the source center wavelength. This is problematic for situations such as cellular imaging, Doppler velocimetry, or polarization sensitive applications where it may be necessary to monitor sample profiles, displacements, phase differences, or refractive index variations which vary rapidly in space or time. A technique previously introduced in phase shifting interferometry uses phase information from multiple wavelengths to overcome this limitation. We show that by appropriate spectral windowing of the broadband light source already used in OCT, particularly by reshaping the source spectrum about two different center wavelengths, the resulting phase variation may be cast in terms of a much longer synthetic wavelength chosen to span the phase variation of interest. We show theoretically that the optimal choice of synthetic wavelength depends upon a tradeoff between the minimum resolvable phase and the length of unambiguous phase measurement. We demonstrate this technique using a broadband source centered at 790 nm by correctly reconstructing the phase profile from a phantom sample containing multiple 2π wrapping artifacts at the center wavelength and compare our result with atomic force microscopy.
We have combined hyperspectral imaging with spectral domain optical coherence tomography (SDOCT) to non-invasively image changes in hemoglobin saturation, blood flow, microvessel morphology and sheer rate on the vessel wall with tumor growth. Changes in these hemodynamic variables were measured over 24 hours in dorsal skin fold window chamber tumors. There was a strong correlation between volumetric flow and hemoglobin saturation (ρ = 0.89, p = 9 × 10-6, N = 15), and a moderate correlation between shear rate on the vessel wall and hemoglobin saturation (ρ = 0.56, p = 0.03, N=15).
KEYWORDS: Optical coherence tomography, Microscopy, Data acquisition, Interferometry, Image processing, Digital holography, Motion measurement, 3D acquisition, Heart, 3D image processing
Spectral domain phase microscopy (SDPM) is a functional extension of optical coherence tomography (OCT) whose
common-path interferometric design enables phase-referenced imaging of dynamic samples. Like OCT, axial resolution
in SDPM is determined by the source coherence length, while lateral resolution is limited by diffraction in the
microscope optics. Nonetheless, the quantitative phase information SDPM generates is sensitive to sub-Angstrom
displacements of scattering structures. Integrative quantitative phase imaging techniques, such as Fourier phase
microscopy, Hilbert phase microscopy, and Digital holographic microscopy, have achieved sub-micron motion detection
in live cells. In contrast with the techniques, SDPM can achieve full depth discrimination, allowing for resolution of the
motion of independent, sub-cellular structures at various cross-sectional planes within the sample. The ability of SDPM
to measure Doppler flow in single-celled organisms, time-resolved cellular motions, and rheological information of the
cytoskeleton has been previously demonstrated. The objective of this study is to extend the use of SDPM to produce
three-dimensional reconstructions of the internal and surface motions of beating cardiomyocytes. Phase information is
used to the motion of quantify cellular structures in the axial dimension. Our gated acquisition process involves
synchronization of the SDPM detection system with and applied electrical field used to stimulate beating in isolated
cardiomyocytes. For a given pacing protocol, we obtain repeat motion measurements in two-dimensions during cellular
contraction, building a volume image by repeating the process at multiple discrete slices through the cell. This
experiment serves as a proof-of-principle for volumetric imaging of beating cardiomyocytes.
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