Michael A. Choma

Sensitivity advantage of swept source and Fourier domain optical coherence tomography

We present theoretical and experimental results which demonstrate the superior sensitivity of swept source (SS) and Fourier domain (FD) optical coherence tomography (OCT) techniques over the conventional time domain (TD) approach. We show that SS- and FD-OCT have equivalent expressions for system signal-to-noise ratio which result in a typical sensitivity advantage of 20-30dB over TD-OCT. Experimental verification is provided using two novel spectral discrimination (SD) OCT systems: a differential fiber-based 800nm FD-OCT system which employs deep-well photodiode arrays, and a differential 1300nm SS-OCT system based on a swept laser with an 87nm tuning range.

Speckle-free laser imaging using random laser illumination

Exploiting the low spatial coherence of specifically designed random lasers, researchers demonstrate speckle-free full-field imaging in the regime of intense optical scattering. Their results show that the quality of images generated from random-laser illumination is superior to those generated from spatially coherent illumination.

Spectral-domain phase microscopy.

Broadband interferometry is an attractive technique for the detection of cellular motions because it provides depth-resolved phase information via coherence gating. We present a phase-sensitive technique called spectral-domain phase microscopy (SDPM). SDPM is a functional extension of spectral-domain optical coherence tomography that allows for the detection of nanometer-scale motions in living cells. The sensitivity of the technique is demonstrated, and its calibration is verified. A shot-noise limit to the displacement sensitivity of this technique is derived. Measurement of cellular dynamics was performed on spontaneously beating cardiomyocytes isolated from chick embryos.

Instantaneous complex conjugate resolved spectral domain and swept-source OCT using 3x3 fiber couplers

We report that the complex conjugate artifact in Fourier domain optical coherence tomography approaches (including spectral domain and swept source OCT) may be resolved by the use of novel interferometer designs based on 3x3 and higher order fiber couplers. Interferometers built from NxN (N>2) truly fused fiber couplers provide simultaneous access to non-complementary phase components of the complex interferometric signal. These phase components may be converted to quadrature components by trigonometric manipulation, then inverse Fourier transformed to obtain A-scans and images with resolved complex conjugate artifact. We demonstrate instantaneous complex conjugate resolved Fourier domain OCT using 3x3 couplers in both spectral domain and swept source implementations. Complex conjugate artifact suppression by factors of ~20dB and ~25dB are demonstrated for spectral domain and swept source implementations, respectively.

Instantaneous quadrature low coherence interferometry with 3 x 3 fiber optic couplers

We present an interferometer topology based on 3 x 3 fiber couplers that gives instantaneous access to the magnitude and phase of die interferometric signal. We demonstrate its performance in heterodyne and homodyne detection with a broadband light source.

Drosophila as a model for the identification of genes causing adult human heart disease.

Drosophila melanogaster genetics provides the advantage of molecularly defined P-element insertions and deletions that span the entire genome. Although Drosophila has been extensively used as a model system to study heart development, it has not been used to dissect the genetics of adult human heart disease because of an inability to phenotype the adult fly heart in vivo. Here we report the development of a strategy to measure cardiac function in awake adult Drosophila that opens the field of Drosophila genetics to the study of human dilated cardiomyopathies. Through the application of optical coherence tomography, we accurately distinguish between normal and abnormal cardiac function based on measurements of internal cardiac chamber dimensions in vivo. Normal Drosophila have a fractional shortening of 87 ± 4%, whereas cardiomyopathic flies that contain a mutation in troponin I or tropomyosin show severe impairment of systolic function. To determine whether the fly can be used as a model system to recapitulate human dilated cardiomyopathy, we generated transgenic Drosophila with inducible cardiac expression of a mutant of human δ-sarcoglycan (δsgS151A), which has previously been associated with familial dilated cardiomyopathy. Compared to transgenic flies overexpressing wild-type δsg, or the standard laboratory strain w1118, Drosophila expressing δsgS151A developed marked impairment of systolic function and significantly enlarged cardiac chambers. These data illustrate the utility of Drosophila as a model system to study dilated cardiomyopathy and the applicability of the vast genetic resources available in Drosophila to systematically study the genetic mechanisms responsible for human cardiac disease.

Swept source optical coherence tomography using an all-fiber 1300-nm ring laser source

The increased sensitivity of spectral domain optical coherence tomography (OCT) has driven the development of a new generation of technologies in OCT, including rapidly tunable, broad bandwidth swept laser sources and spectral domain OCT interferometer topologies. In this work, the operation of a turnkey 1300-nm swept laser source is demonstrated. This source has a fiber ring cavity with a semiconductor optical amplifier gain medium. Intracavity mode selection is achieved with an in-fiber tunable fiber Fabry-Perot filter. A novel optoelectronic technique that allows for even sampling of the swept source OCT signal in k space also is described. A differential swept source OCT system is presented, and images of in vivo human cornea and skin are presented. Lastly, the effects of analog-to-digital converter aliasing on image quality in swept source OCT are discussed.

Optical Coherence Tomography A New High-Resolution Imaging Technology to Study Cardiac Development in Chick Embryos

Background—Optical coherence tomography (OCT) is a depth-resolved, noninvasive, non-destructive imaging modality, the use of which has yet to be fully realized in developmental biology. Methods and Results—We visualized embryonic chick hearts at looping stages using an OCT system with a 22 &mgr;m axial and 27 &mgr;m lateral resolution and an acquisition rate of 4000 A-scans per second. Normal chick embryos from stages 14 to 22 and sham-operated and cardiac neural crest-ablated embryos from stages 15 and 18 were scanned by OCT. Three-dimensional data sets were acquired and processed to create volumetric reconstructions and short video clips. The OCT-scanned embryos (2 in each group) were photographed after histological sectioning in comparable planes to those visualized by OCT. The optical and histological results showing cardiovascular microstructures such as myocardium, the cardiac jelly, and endocardium are presented. Conclusions—OCT is a powerful imaging modality which can provide new insight in assessing and understanding normal and abnormal cardiac development in a variety of animal models.

Molecular contrast in optical coherence tomography by use of a pump–probe technique

We describe a novel technique for contrast enhancement in optical coherence tomography (OCT) that makes possible molecular-specific imaging for what is believed to be the first time. A pump-probe technique is employed in which a pulsed pump laser is tuned to ground-state absorption in a molecule of interest. The location of the target molecule population is derived from the resulting transient absorption of OCT sample-arm light acting as probe light. A signal processing technique for three-dimensional localization of the transient absorption signal is described, and preliminary results exhibiting OCT contrast from methylene blue dye in multilayer and scattering phantoms are presented.

Heterodyne swept-source optical coherence tomography for complete complex conjugate ambiguity removal.

Fourier domain (FD) techniques have increasingly gained attention in optical coherence tomography (OCT). This is primarily due to their demonstrated sensitivity of two to three orders of magnitude over conventional time-domain techniques. FDOCT images are subject to two primary sources of artifacts. First, a complex conjugate ambiguity arises because the Fourier transform of the real-valued spectral interferometric signal is Hermitian symmetric. This ambiguity leads to artifactual superposition of reflectors at positive and negative pathlength differences between the sample and reference reflectors. Second, noninterferometric and sample autocorrelation terms appear at dc, obscuring reflectors at zero pathlength difference. We show that heterodyne detection in swept-source OCT (SSOCT) enables the resolution of complex conjugate ambiguity and the removal of noninterferometric and autocorrelation artifacts. We also show that complex conjugate ambiguity resolution via frequency shifting circumvents fall-off induced by finite source linewidth in SSOCT when samples are shifted to large pathlength differences. We describe an efficient heterodyne SSOCT design that enables compensation of power losses from frequency-shifting elements. Last, we demonstrate this technique, coupled with wavenumber triggering and electronic demodulation, for in vivo imaging of the human anterior eye segment.