Mingtao Zhao

Adaptive-optics optical coherence tomography for high-resolution and high-speed 3D retinal in vivo imaging

We have combined Fourier-domain optical coherence tomography (FD-OCT) with a closed-loop adaptive optics (AO) system using a Hartmann-Shack wavefront sensor and a bimorph deformable mirror. The adaptive optics system measures and corrects the wavefront aberration of the human eye for improved lateral resolution (~4 μm) of retinal images, while maintaining the high axial resolution (~6 μm) of stand alone OCT. The AO-OCT instrument enables the three-dimensional (3D) visualization of different retinal structures in vivo with high 3D resolution (4×4×6 μm). Using this system, we have demonstrated the ability to image microscopic blood vessels and the cone photoreceptor mosaic.

High-speed complex conjugate resolved retinal spectral domain optical coherence tomography using sinusoidal phase modulation.

We demonstrate high-speed complex conjugate artifact (CCA) resolved imaging of human retina in vivo using spectral domain optical coherence tomography. This technique utilizes sinusoidal reference mirror modulation to implement high-speed integrating buckets acquisition and a quadrature projection reconstruction algorithm in postprocessing. This method is illustrated experimentally using sets of four integrating bucket phase scans, acquired at 52 kHz, for DC suppression of 73 dB and complex conjugate suppression of 35 dB. Densely sampled (3000 A-scans/image, acquired at 4.3 images/s) full-depth in vivo images of optic nerve head show CCA suppression for most image reflections to the noise floor.

3D refraction correction and extraction of clinical parameters from spectral domain optical coherence tomography of the cornea

Capable of three-dimensional imaging of the cornea with micrometer-scale resolution, spectral domain-optical coherence tomography (SDOCT) offers potential advantages over Placido ring and Scheimpflug photography based systems for accurate extraction of quantitative keratometric parameters. In this work, an SDOCT scanning protocol and motion correction algorithm were implemented to minimize the effects of patient motion during data acquisition. Procedures are described for correction of image data artifacts resulting from 3D refraction of SDOCT light in the cornea and from non-idealities of the scanning system geometry performed as a pre-requisite for accurate parameter extraction. Zernike polynomial 3D reconstruction and a recursive half searching algorithm (RHSA) were implemented to extract clinical keratometric parameters including anterior and posterior radii of curvature, central cornea optical power, central corneal thickness, and thickness maps of the cornea. Accuracy and repeatability of the extracted parameters obtained using a commercial 859nm SDOCT retinal imaging system with a corneal adapter were assessed using a rigid gas permeable (RGP) contact lens as a phantom target. Extraction of these parameters was performed in vivo in 3 patients and compared to commercial Placido topography and Scheimpflug photography systems. The repeatability of SDOCT central corneal power measured in vivo was 0.18 Diopters, and the difference observed between the systems averaged 0.1 Diopters between SDOCT and Scheimpflug photography, and 0.6 Diopters between SDOCT and Placido topography.

Synthetic wavelength based phase unwrapping in spectral domain optical coherence tomography

Phase sensing implementations of spectral domain optical coherence tomography (SDOCT) have demonstrated the ability to measure nanometer-scale temporal and spatial profiles of samples. However, the phase information suffers from a 2pi ambiguity that limits observations of larger sample displacements to lengths less than half the source center wavelength. We introduce a synthetic wavelength phase unwrapping technique in SDOCT that uses spectral windowing and corrects the 2pi ambiguity, providing accurate measurements of sample motion with information gained from standard SDOCT processing. We demonstrate this technique by using a common path implementation of SDOCT and correctly measure phase profiles from a phantom phase object and human epithelial cheek cells which produce multiple wrapping artifacts. Using a synthetic wavelength for phase unwrapping could prove useful in Doppler or other phase based implementations of OCT.

Real-time spectral domain Doppler optical coherence tomography and investigation of human retinal vessel autoregulation

Investigation of the autoregulatory mechanism of human retinal perfusion is conducted with a real-time spectral domain Doppler optical coherence tomography (SDOCT) system. Volumetric, time-sequential, and Doppler flow imaging are performed in the inferior arcade region on normal healthy subjects breathing normal room air and 100% oxygen. The real-time Doppler SDOCT system displays fully processed, high-resolution [512 (axial) x 1000 (lateral) pixels] B scans at 17 frames/sec in volumetric and time-sequential imaging modes, and also displays fully processed overlaid color Doppler flow images comprising 512 (axial) x 500 (lateral) pixels at 6 frames/sec. Data acquired following 5 min of 100% oxygen inhalation is compared with that acquired 5 min postinhalation for four healthy subjects. The average vessel constriction across the population is -16+/-26% after oxygen inhalation with a dilation of 36+/-54% after a return to room air. The flow decreases by -6+/-20% in response to oxygen and in turn increases by 21+/-28% as flow returns to normal in response to room air. These trends are in agreement with those previously reported using laser Doppler velocimetry to study retinal vessel autoregulation. Doppler flow repeatability data are presented to address the high standard deviations in the measurements.

Single-camera sequential-scan-based polarization-sensitive SDOCT for retinal imaging

A single-camera, high-speed, polarization-sensitive, spectral-domain optical-coherence-tomography system was developed to measure the polarization properties of the in vivo human retina. A novel phase-unwrapping method in birefringent media is described to extract the total reflectivity, accumulative retardance, and fast-axis orientation from a specially designed sequence of polarization states incident on the sample. A quarter-wave plate was employed to test the performance of the system. The average error and standard deviation of retardation measurements were 3.2 degrees and 2.3 degrees , respectively, and of the fast-axis orientation 1.2 degrees and 0.7 degrees over the range of 0 degrees -180 degrees . The depolarization properties of the retinal pigment epithelium were clearly observed in both retardance and fast-axis orientation image. A normalized standard deviation of the retardance and of the fast-axis orientation is introduced to segment the polarization-scrambling layer of the retinal pigment epithelium.

Synthetic wavelength-based phase unwrapping in Fourier domain optical coherence tomography

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.

3D OCT imaging in clinical settings : toward quantitative measurements of retinal structures

The acquisition speed of current FD-OCT (Fourier Domain - Optical Coherence Tomography) instruments allows rapid screening of three-dimensional (3D) volumes of human retinas in clinical settings. To take advantage of this ability requires software used by physicians to be capable of displaying and accessing volumetric data as well as supporting post processing in order to access important quantitative information such as thickness maps and segmented volumes. We describe our clinical FD-OCT system used to acquire 3D data from the human retina over the macula and optic nerve head. B-scans are registered to remove motion artifacts and post-processed with customized 3D visualization and analysis software. Our analysis software includes standard 3D visualization techniques along with a machine learning support vector machine (SVM) algorithm that allows a user to semi-automatically segment different retinal structures and layers. Our program makes possible measurements of the retinal layer thickness as well as volumes of structures of interest, despite the presence of noise and structural deformations associated with retinal pathology. Our software has been tested successfully in clinical settings for its efficacy in assessing 3D retinal structures in healthy as well as diseased cases. Our tool facilitates diagnosis and treatment monitoring of retinal diseases.

Exposure time dependence of image quality in high-speed retinal in vivo Fourier domain OCT

We built a Fourier domain optical coherence tomography (FD-OCT) system using a line scan CCD camera that allows real time data display and acquisition. This instrument is able to produce 2D B-scans as well as 3D data sets with human subjects in vivo in clinical settings. In this paper we analyze the influence of varying exposure times of the CCD detector on image quality. Sensitivity values derived from theoretical predictions have been compared with measurements (obtained with mirrors and neutral density filters placed in both interferometer arms). The results of these experiments, discussion about differences between sensitivity values, potential sources of discrepancies, and recommendations for optimal exposure times will be described in this paper. A short discussion of observed artifacts as well as possible ways to remove them is presented. The influence of relative retinal position with respect to reference mirror position will also be described.

Corneal biometry from volumetric SDOCT and comparison with existing clinical modalities

We present a comparison of corneal biometric values from dense volumetric spectral domain optical coherence tomography (SDOCT) scans to reference values in both phantoms and clinical subjects. We also present a new optically based “keratometric equivalent power” formula for SDOCT that eliminates previously described discrepancies between corneal power form SDOCT and existing clinical modalities. Phantom objects of varying radii of curvature and corneas of normal subjects were imaged with a clinical SDOCT system. The optically corrected three-dimensional surfaces were used to recover radii of curvature and power as appropriate. These were then compared to the manufacturer’s reference values in phantoms and to measurements from topography and Scheimpflug photography in subjects. In phantom objects, paired differences between SDOCT and reference values for radii of curvature were not statistically significant. In subjects, there were no significant paired differences between SDOCT and reference values from the other modalities for anterior radius and corneal keratometric power. In contrast to other studies, we found that dense volumetric scans with available SDOCT can be used to recover corneal biometric values—including power—that correspond well with existing clinical measurements.