Supercontinuum light source enables in vivo optical microangiography of capillary vessels within tissue beds

This Letter reports on the use of a supercontinuum light source to achieve ultrahigh resolution and ultrahigh sensitive optical microangiography (OMAG) imaging of microcirculations within tissue beds in vivo. After passing through a specially designed optical filter with a passband of 120 nm centered on 800 nm, the light source is coupled into an optic-fiber-based OMAG system that provides a measured axial resolution of ～3 μm over a ranging distance of 2 mm. Within this ranging distance, the system gives an averaged signal-to-noise ratio of 87 dB and a sensitivity roll-off of 7 dB at an A-scan rate of 70 kHz. We demonstrate the capability of the system to visualize a detailed microvascular perfusion map, including the single red blood cells within the capillaries, by imaging a mouse ear flap in vivo.     

Imaging of subcutaneous blood vessels and flow velocity profiles by optical coherence tomography

We have applied a compact low power rapid scanning Doppler Optical Coherence Tomography system to monitor multi-dimensional velocity profiles within the complex vessels and simultaneous real-time non-invasive imaging of skin tissues morphology in vivo, in the wavelength range of 1.3–1.5 nm. Optical clearing of skin tissues has been utilized to achieve depth of OCT images up to 1.7 mm. Current approach enables applying low-power (0.4–0.5 mW) and low-noise broadband near-infrared light sources and obtaining OCT images with down to 12 μm spatial resolution. Two-dimensional time-domain OCT images of complex flow velocity profiles in blood vessel phantom and in vivo subcutaneous human skin tissues are presented. The effect of optical clearing on in vivo images is demonstrated and discussed.     

Label-free 3D imaging of microstructure, blood, and lymphatic vessels within tissue beds in vivo

This Letter reports the use of an ultrahigh resolution optical microangiography (OMAG) system for simultaneous 3D imaging of microstructure and lymphatic and blood vessels without the use of an exogenous contrast agent. An automatic algorithm is developed to segment the lymphatic vessels from the microstructural images based on the fact that the lymph fluid is optically transparent. An OMAG system is developed that utilizes a broadband supercontinuum light source, providing an axial resolution of 2.3 μm and lateral resolution of 5.8 μm, capable of resolving the capillary vasculature and lymphatic vessels innervating microcirculatory tissue beds. Experimental demonstration is performed by showing detailed 3D lymphatic and blood vessel maps, coupled with morphology, within mouse ears in vivo.     

Arbitrary Three-Phase Shifting Algorithm for Achieving Full Range Spectral Optical Coherence Tomography

An arbitrary three-phase shifting algorithm is introduced in order to achieve full range spectral optical coherence tomography imaging of biological tissue. Theoretical treatment behind this approach is given and experimentally verified. It is shown that this method is capable of eliminating the undesired auto-correlation and complex conjugate images, leading to the un-obscured full range spectral OCT imaging. An intact porcine eye is used to demonstrate the potential of such a method for biological imaging.     

Interstitial Doppler optical coherence tomography

Doppler optical coherence tomography (OCT) can image tissue structure and blood flow at micrometer-scale resolution but has limited imaging depth. We report a novel, linear-scanning, needle-based Doppler OCT system using angle-polished gradient-index or ball-lensed fibers. A prototype system with a 19-guage (diameter of approximately 0.9 mm) echogenic needle is constructed and demonstrates in vivo imaging of bidirectional blood flow in rat leg and abdominal cavity. To our knowledge, this is the first demonstration of Doppler OCT through a needle probe in interstitial applications to visualize deeply situated microcirculation.     

Directional blood flow imaging in volumetric optical microangiography achieved by digital frequency modulation

An effective digital frequency modulation approach to achieve directional blood flow imaging within microcirculations in tissue beds in vivo for optical microangiography is presented. The method only requires the system to capture one three-dimensional data set within which the interferograms are modulated by a constant frequency modulation that gives one directional flow information. The result is that the imaging speed is doubled and the computational load is halved. The method is experimentally validated by a flow phantom and is tested for imaging of cerebral vascular blood perfusion in a live mouse with the cranium left intact.     

In vivo bidirectional color Doppler flow imaging of picoliter blood volumes using optical coherence tomography

We describe a novel optical system for bidirectional color Doppler imaging of flow in biological tissues with micrometer-scale resolution and demonstrate its use for in vivo imaging of blood flow in an animal model. Our technique, color Doppler optical coherence tomography (CDOCT), performs spatially localized optical Doppler velocimetry by use of scanning low-coherence interferometry. CDOCT is an extension of optical coherence tomography (OCT), employing coherent signal-acquisition electronics and joint time-frequency analysis algorithms to perform flow imaging simultaneous with conventional OCT imaging. Cross-sectional maps of blood flow velocity with <50-microm spatial resolution and <0.6-mm/s velocity precision were obtained through intact skin in living hamster subdermal tissue. This technology has several potential medical applications.     

Use of a scanner to modulate spatial interferograms for in vivo full-range Fourier-domain optical coherence tomography

We report a new yet simple method to achieve full-range complex Fourier-domain optical coherence tomography (OCT) for in vivo imaging. The method utilizes a scanner that is dedicated for lateral scanning in the system to introduce a constant carrier frequency into the OCT spectral interferograms during the scanning. This is achieved by simply offsetting the sampling beam spot away from the pivot point of the scanning mirror. We demonstrate the method experimentally for in vivo full-range imaging of the anterior segment of a human eye. The method is free from complex conjugate mirror image and self-cross-correlation image artifacts.     

幅值和相位配准技术及其在光学相干层析血流成像中的应用

频域光学相干层析系统中扫描机构定位精度、 机械抖动及样品移动会造成A扫描信号幅值和相位发生波动, 影响生物组织成像质量. 利用最小灰度差匹配、Lorentzian曲线极值拟合和谱域光程差补偿等方法对A 扫描信号进行幅值配准. 通过对A扫描信号相位分布特征的匹配实现相位差检测与配准. 通过求已配准的A 扫描复信号之差, 消除静态组织对血流成像的影响. 进行了人眼扫描实验, 有效提取了视网膜三维血流图像. 实验结果表明, 提出的幅值及相位配准方法大大减小了系统扫描精度、人眼跳动等因素对生物组织在体成像质量的影响. 快速、精确的相位配准方法也可广泛应用在多普勒OCT、相位显微等与相位分辨有关的光学成像领域. 关键词： 频域光学相干层析 配准 血流成像 相位分布特征     

Combined Raman spectroscopy and optical coherence tomography device for tissue characterization

We report a dual-modal device capable of sequential acquisition of Raman spectroscopy (RS) and optical coherence tomography (OCT) along a common optical axis. The device enhances application of both RS and OCT by precisely guiding RS acquisition with OCT images while also compensating for the lack of molecular specificity in OCT with the biochemical specificity of RS. We characterize the system performance and demonstrate the capability to identify structurally ambiguous features within an OCT image with RS in a scattering phantom, guide acquisition of RS from a localized malignancy in ex vivo breast tissue, and perform in vivo tissue analysis of a scab.     

Full range spectral domain optical coherence tomography using a fiber-optic probe as a self-phase shifter

We present a full range handheld probe type spectral domain optical coherence tomography (SD-OCT) method. Here, the sample arm is composed of a tilted fiber-optic cantilever scanner; thus, the phase shift concurrently occurs while sample scanning. With the phase shift, we could achieve a full range complex-conjugate-free OCT image with no additional phase shifters in the reference arm. To realize this technique, a magnetically actuated probe was adopted. Full range SD-OCT images of a pearl, human fingernail, and human tooth were subsequently obtained using this suggested probe. The scanning range and acquisition speed were 3?mm and 20 frames/s?, respectively.     

Real-time in vivo blood-flow imaging by moving-scatterer-sensitive spectral-domain optical Doppler tomography

We present a moving-scatterer-sensitive optical Doppler tomography (MSS-ODT) technique for in vivo blood flow imaging in real time by using a spectral-domain optical coherence tomography system. In MSS-ODT the influence of stationary scatterers is suppressed by subtracting adjacent complex axial scans before calculating the Doppler frequency shift. We demonstrate that MSS-ODT is a useful technique for accurate determination of blood vessel size by imaging flow in a small capillary tube with a 75 microm inner diameter. The flow profile obtained with MSS-ODT yields a substantially more accurate tube diameter than that obtained with the conventional phase-resolved method, which underestimates the diameter by about 23%. We also demonstrate that MSS-ODT provides improved sensitivity over the conventional phase-resolved method for imaging in vivo blood flow in small vessels in a mouse ear.     

Ultrahigh-resolution optical coherence tomography using continuum generation in an air-silica microstructure optical fiber

We demonstrate ultrahigh-resolution optical coherence tomography (OCT) using continuum generation in an air-silica microstructure fiber as a low-coherence light source. A broadband OCT system was developed and imaging was performed with a bandwidth of 370 nm at a 1.3-mu;m center wavelength. Longitudinal resolutions of 2.5 microm in air and ~2 microm in tissue were achieved. Ultrahigh-resolution imaging in biological tissue in vivo was demonstrated.     

Intraluminal fiber-optic Doppler imaging catheter for structural and functional optical coherence tomography

We describe a miniature fiber-optic Doppler imaging catheter for integrated functional and structural optical coherence tomography (OCT) imaging. The Doppler catheter can map blood flow within a vessel as well as image vessel wall structures. A prototype Doppler catheter has been developed and demonstrated for measuring the intraluminal velocity profile in a vessel phantom (conduit). A simple mathematical model is demonstrated to estimate the total flow rate. This estimation technique also enables the spatial range of flow measurements to be extended by approximately two times the normal OCT image-penetration depth. The Doppler OCT catheter could be a powerful device for cardiovascular imaging.     

Spectroscopic optical coherence tomography

Spectroscopic optical coherence tomography (OCT), an extension of conventional OCT, is demonstrated for performing cross-sectional tomographic and spectroscopic imaging. Information on the spectral content of backscattered light is obtained by detection and processing of the interferometric OCT signal. This method allows the spectrum of backscattered light to be measured over the entire available optical bandwidth simultaneously in a single measurement. Specific spectral features can be extracted by use of digital signal processing without changing the measurement apparatus. An ultrabroadband femtosecond Ti:Al(2)O(3) laser was used to achieve spectroscopic imaging over the wavelength range from 650 to 1000 nm in a simple model as well as in vivo in the Xenopus laevis (African frog) tadpole. Multidimensional spectroscopic data are displayed by use of a novel hue-saturation false-color mapping.     

Noninvasive imaging of in vivo blood flow velocity using optical Doppler tomography

We report the development of an optical technique for noninvasive imaging of in vivo blood flow dynamics and tissue structures with high spatial resolution (2-10 microm) in biological systems. The technique is based on optical Doppler tomography (ODT), which combines Doppler velocimetry with optical coherence tomography to measure blood flow velocity at discrete spatial locations. The exceptionally high resolution of ODT permits noninvasive in vivo imaging of both blood microcirculation and tissue structures surrounding the vessel, which has significance for biomedical research and clinical applications. Tomographic imaging of in vivo blood flow velocity in the chick chorioallantoic membrane and in rodent skin is demonstrated.     

Principal-component-analysis-based estimation of blood flow velocities using optical coherence tomography intensity signals

The intensity signal in optical coherence tomography contains information about the translational velocity of scatterers, and can be used to quantify blood flow. We apply principal component analysis to efficiently extract this information. We also study use of nonuniform temporal sampling of the intensity signal to increase the range of quantifiable flow velocities. We demonstrate this technique in simulation, phantom and in vivo blood flow measurements, and highlight its potential to enable three-dimensional wide-field mapping of blood flow using OCT.     

High-speed, high-resolution optical coherence tomography retinal imaging with a frequency-swept laser at 850 nm

High-speed, high-resolution optical coherence tomography (OCT) imaging of the human retina is demonstrated using a frequency-swept laser at 850 nm. A compact external cavity semiconductor laser design, optimized for swept-source ophthalmic OCT, is described. The laser enables an effective 16 kHz sweep rate with >10 mm coherence length and a tuning range of approximately 35 nm full width at half-maximum, yielding an axial resolution of <7 micro m in tissue.     

Extraction of optical scattering parameters and attenuation compensation in optical coherence tomography images of multilayered tissue structures

A recently developed analytical optical coherence tomography (OCT) model [Thrane et al., J. Opt. Soc. Am. A 17, 484 (2000)] allows the extraction of optical scattering parameters from OCT images, thereby permitting attenuation compensation in those images. By expanding this theoretical model, we have developed a new method for extracting optical scattering parameters from multilayered tissue structures in vivo. To verify this, we used a Monte Carlo (MC) OCT model as a numerical phantom to simulate the OCT signal for heterogeneous multilayered tissue. Excellent agreement between the extracted values of the optical scattering properties of the different layers and the corresponding input reference values of the MC simulation was obtained, which demonstrates the feasibility of the method for in vivo applications. This is to our knowledge the first time such verification has been obtained, and the results hold promise for expanding the functional imaging capabilities of OCT.     

Speckle variance detection of microvasculature using swept-source optical coherence tomography

We report on imaging of microcirculation by calculating the speckle variance of optical coherence tomography (OCT) structural images acquired using a Fourier domain mode-locked swept-wavelength laser. The algorithm calculates interframe speckle variance in two-dimensional and three-dimensional OCT data sets and shows little dependence to the Doppler angle ranging from 75 degrees to 90 degrees . We demonstrate in vivo detection of blood flow in vessels as small as 25 microm in diameter in a dorsal skinfold window chamber model with direct comparison with intravital fluorescence confocal microscopy. This technique can visualize vessel-size-dependent vascular shutdown and transient vascular occlusion during Visudyne photodynamic therapy and may provide opportunities for studying therapeutic effects of antivascular treatments without on exogenous contrast agent.