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.     

Doppler standard deviation imaging for clinical monitoring of in vivo human skin blood flow

We used a novel phase-resolved optical Doppler tomographic (ODT) technique with very high flow-velocity sensitivity (10microm/s) and high spatial resolution (10microm) to image blood flow in port-wine stain (PWS) birthmarks in human skin. In addition to the regular ODT velocity and structural images, we use the variance of blood flow velocity to map the PWS vessels. Our device combines ODT and therapeutic systems such that PWS blood flow can be monitored in situ before and after laser treatment. To the authors' knowledge this is the first clinical application of ODT to provide a fast semiquantitative evaluation of the efficacy of PWS laser therapy in situ and in real time.     

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.     

光学相干层析多普勒成像功能拓展研究

光学多普勒成像(Optical Doppler tomography,ODT)是一种结合了光学相干层析成像技术(Opticalcoherence tomography,OCT)和多普勒流速仪的非侵入、非接触的成像技术,能够实现对高散介质组织内部的血管分布和血液流速的探测。阐述了基于数字希尔伯特变换的相位分离多普勒光学相干层析成像技术的工作原理,并且通过对玻璃毛细管和生物芯片微通道管中聚苯乙烯溶液流速的实验测量,准确测量管内微粒缓慢移动时的多普勒频移量,获得了玻璃管内和生物芯片微通道管中流速分布曲线,证实了所提方法的可行性。获取的多普勒图像具有较高的空间分辨力和速度分辨力,在未来的临床应用中有潜在的应用价值。     

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.     

Real-time phase-resolved functional optical coherence tomography by use of optical Hilbert transformation

We have developed a novel real-time phase-resolved functional optical coherence tomography system that uses optical Hilbert transformation. When we use a resonant scanner in the reference arm of the interferometer, with an axial scanning speed of 4 kHz, the frame rate of both structural and Doppler blood-flow imaging with a size of 100 by 100 pixels is 10 Hz. The system has high sensitivity and a larger dynamic range for measuring the Doppler frequency shift that is due to moving red blood cells. Real-time images of in vivo blood flow in human skin obtained with this interferometer are presented.     

Polarization-resolved second-harmonic-generation optical coherence tomography in collagen

We describe a novel imaging technique, second-harmonic-generation optical coherence tomography (SHOCT). This technique combines the spatial resolution and depth penetration of optical coherence tomography (OCT) with the molecular sensitivity of second-harmonic-generation spectroscopy. As a consequence of the coherent detection required for OCT, polarization-resolved images arise naturally. We demonstrate this new technique on a skin sample from the belly of Icelandic salmon, acquiring polarization-resolved SHOCT and OCT images simultaneously.     

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.     

Real-time flow imaging by removing texture pattern artifacts in spectral-domain optical Doppler tomography

We present a new, simple method to suppress texture pattern artifacts induced by the optical heterogeneity of tissues to improve the performance of flow imaging for real-time phase-resolved optical Doppler tomography. The method performs transverse scanning of the probe beam in the forward and then reverse directions, and it takes average of the spatial phase changes between them to obtain the final velocity image. It relies on the fact that the phase changes between successive axial scans due to the optical heterogeneity of the sample are time independent, while those due to the moving particles are time dependent. We experimentally demonstrate this method by real-time imaging of a flow phantom.     

Tracking optical coherence tomography

An experimental tracking optical coherence tomography (OCT) system has been clinically tested. The prototype instrument uses a secondary sensing beam and steering mirrors to compensate for eye motion with a closed-loop bandwidth of 1 kHz and tracking accuracy, to within less than the OCT beam diameter. The retinal tracker improved image registration accuracy to <1 transverse pixel (<60 microm). Composite OCT images averaged over multiple scans and visits show a sharp fine structure limited only by transverse pixel size. As the resolution of clinical OCT systems improves, the capability to reproducibly map complex structures in the living eye at high resolution will lead to improved understanding of disease processes and improved sensitivity and specificity of diagnostic procedures.     

In vivo measurement of retinal physiology with high-speed ultrahigh-resolution optical coherence tomography

Noninvasive in vivo functional optical imaging of the intact retina is demonstrated by using high-speed, ultrahigh-resolution optical coherence tomography (OCT). Imaging was performed with 2.8 microm resolution at a rate of 24,000 axial scans per second. A white-light stimulus was applied to the dark-adapted rat retina, and the average reflectivities from different intraretinal layers were monitored as a function of time. A 10%-15% increase in the average amplitude reflectance of the photoreceptor outer segments was observed in response to the stimulus. The spatial distribution of the change in the OCT signal is consistent with an increase in backscatter from the photoreceptor outer segments. To our knowledge, this is the first in vivo demonstration of OCT functional imaging in the intact retina.     

Fourier domain optical coherence tomography using optical demultiplexers imaging at 60,000,000 lines/s

We describe high-speed Fourier domain optical coherence tomography (OCT) using optical demultiplexers (ODs) for spectral dispersion. The OD enables separation of a narrow spectral band of 14 GHz (0.11 nm) from a broadband incident light at 256 different frequencies in 25.0 GHz intervals centered at 192.2 THz (1559.8 nm). OCT imaging of 60,000,000 axial scans per second was achieved through parallel signal acquisition using 256 balanced photoreceivers to simultaneously detect all the output signals from the ODs in a Fourier domain OCT system. OCT imaging at a 16 kHz frame rate, 1100 A-lines per frame, 3 mm depth range, and 23 microm resolution was demonstrated using a resonant scanner for lateral scanning.     

Holoscopy--holographic optical coherence tomography

Scanning optical coherence tomography (OCT) is limited in sensitivity and resolution by the restricted focal depth of the confocal detection scheme. Holoscopy, a combination of holography and Fourier-domain full-field OCT, is proposed as a way to detect photons from all depths of a sample volume simultaneously with uniform sensitivity and lateral resolution, even at high NAs. By using the scalar diffraction theory, as frequently applied in digital holographic imaging, we fully reconstruct the object field with depth-invariant imaging quality. In vivo imaging of human skin is demonstrated with an image quality comparable to conventionally scanned OCT.     

Birefringence imaging of human skin by polarization-sensitive spectral interferometric optical coherence tomography

We have developed a spectral interferometric optical coherence tomography (OCT) system with polarization sensitivity that is able to measure a two-dimensional tomographic image by means of one-dimensional mechanical scanning. Our system, which has an axial resolution of 32 mum , calculates the distribution of each element of the Müller matrix of a measured object from 16 OCT images. The OCT system successfully reveals the birefringent nature of human skin tissue.     

Fiber-optic-bundle-based optical coherence tomography

A fiber-optic-bundle-based optical coherence tomography (OCT) probe method is presented. The experimental results demonstrate this multimode optical fiber-bundle-based OCT system can achieve a lateral resolution of 12 microm and an axial resolution of 10 microm with a superluminescent diode source. This novel OCT imaging approach eliminates any moving parts in the probe and has a primary advantage for use in extremely compact and safe OCT endoscopes for imaging internal organs and great potential to be combined with confocal endoscopic microscopy.     

In vivo human retinal imaging by ultrahigh-speed spectral domain optical coherence tomography

An ultrahigh-speed spectral domain optical coherence tomography (SD-OCT) system is presented that achieves acquisition rates of 29,300 depth profiles/s. The sensitivity of SD-OCT and time domain OCT (TD-OCT) are experimentally compared, demonstrating a 21.7-dB improvement of SD-OCT over TD-OCT. In vivo images of the human retina are presented, demonstrating the ability to acquire high-quality structural images with an axial resolution of 6 microm at ultrahigh speed and with an ocular exposure level of less than 600 microW.     

Phase-resolved functional optical coherence tomography: simultaneous imaging of in situ tissue structure, blood flow velocity, standard deviation, birefringence, and Stokes vectors in human skin

We describe a phase-resolved functional optical coherence tomography system that can simultaneously yieldin situ images of tissue structure, blood flow velocity, standard deviation, birefringence, and the Stokes vectors in human skin. Multifunctional images were obtained by processing of analytical interference fringe signals derived from two perpendicular polarization-detection channels. The blood flow velocity and standard deviation images were obtained by comparison of the phases from pairs of analytical signals in neighboring A-lines in the same polarization state. The analytical signals from two polarization-diversity detection channels were used to determine the four Stokes vectors for four reference polarization states. From the four Stokes vectors, the birefringence image, which is not sensitive to the orientation of the optical axis in the sample, was obtained. Multifunctional in situ images of a port wine stain birthmark in human skin are presented.     

Homodyne en face optical coherence tomography

We demonstrate, for what we believe to be the first time, the use of a 3 x 3 fiber-optic coupler to realize a homodyne optical coherence tomography (OCT) system for en face imaging of highly scattering tissues and turbid media. The homodyne OCT setup exploits the inherent phase shifts between different output ports of a 3 x 3 fiber-optic coupler to extract amplitude information of a sample. Our homodyne en face OCT system features a measured resolution of 14 microm axially and 9.4 microm laterally with a 90 dB signal-to-noise ratio at 10 micros integration time. En face OCT imaging of a stage 52 Xenopus laevis was successfully demonstrated at a depth of 600 microm within the sample.     

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     

光学多普勒层析三维矢量测速方法研究

光学多普勒层析术(ODT)是一种高分辨、非侵入的生物医学成像手段,能同时得到组织的结构信息和组织内血管的流速信息.提出了一种新型的基于相位分辨技术的ODT三维矢量测速方法.在ODT系统样品臂的准直镜和聚焦透镜之间加入窄带相位片,形成三个不同的相位延迟,通过计算多普勒频移和不同相位延迟下的多普勒展宽,可得到毛细管内的三维矢量流场分布.对已知浓度的聚苯乙烯溶液进行了一系列不同角度和不同流速的实验,结果证明这种新型的ODT矢量测速方法可以较精确的实现三维矢量流速的测量.