Flow Assessment Using Optical Coherence Microscopy Based Particle Image Velocimetry

Congenital heart diseases (CHDs) are the most common forms of congenital malformation in newborns. Among all types of CHDs, a large portion is contributed by malformation of endocardial cushion malformation during early heart development. Although the etiology of endocardial cushion malformation is unclear, it is a result of interactions between genetic and environmental factors has been confirmed. There is hypothesis indicating that malformation of endocardial cushion is caused by altered shear stress conditions where in cushion forming area the shear stress is supposed to be high compare with other area in congenital heart. However it is difficult to justify due to lack of in vivo imaging modality that is able to monitor structure and hemodynamic conditions simultaneously and over long time period. To address this problem, we present an optical coherence microscopy based particle image velocimetry system. This system is capable of invasively imaging biological sample structures at micrometer resolution and providing velocity information at the same time. With this imaging set up we successfully assessed velocity profile in a microfluidic system with simultaneous structure details demonstration of the microfluidic channel. Both flow measurement and structural information were verified using conventional microscopy. As a result, OCM-based PIV imaging modality not only makes it feasible to study in detail the process of congenital heart remodeling in response to environmental alterations, but also provides new options for measuring fluid flow in live tissue

Development of High-speed Optical Coherence Tomography for Time-lapse Non-destructive Characterization of Samples

Optical coherence tomography (OCT) is an established optical imaging modality which can obtain label-free, non-destructive 3D images of samples with micron-scale resolution and millimeter penetration. OCT has been widely adopted for biomedical researches

Doppler imaging with dual-detection full-range frequency domain optical coherence tomography

Most of full-range techniques for Frequency Domain Optical Coherence Tomography (FD-OCT) reported to date utilize the phase relation between consecutive axial lines to reconstruct a complex interference signal and hence may exhibit degradation in either mirror image suppression performance or detectable velocity dynamic range or both when monitoring a moving sample such as flow activity. We have previously reported a technique of mirror image removal by simultaneous detection of the quadrature components of a complex spectral interference called a Dual-Detection Frequency Domain OCT (DD-FD-OCT) [Opt. Lett. 35, 1058-1060 (2010)]. The technique enables full range imaging without any loss of acquisition speed and is intrinsically less sensitive to phase errors generated by involuntary movements of the subject. In this paper, we demonstrate the application of the DD-FD-OCT to a phase-resolved Doppler imaging without degradation in either mirror image suppression performance or detectable velocity dynamic range that were observed in other full-range Doppler methods. In order to accommodate for Doppler imaging, we have developed a fiber-based DD-FD-OCT that more efficiently utilizes the source power compared with the previous free-space DD-FD-OCT. In addition, the velocity sensitivity of the phase-resolved DD-FD-OCT was investigated, and the relation between the measured Doppler phase shift and set flow velocity of a flow phantom was verified. Finally, we demonstrate the Doppler imaging using the DD-FD-OCT in a biological sample

Swept-source based, single-shot, multi-detectable velocity range Doppler optical coherence tomography

Phase-Resolved Doppler Optical Coherence Tomography (PR-DOCT) allows visualization and characterization of the location, direction, velocity, and profile of flow activity embedded in a static sample structure. The detectable Velocity Dynamic Range (VDR) of each particular PR-DOCT system is governed by a detectable Doppler phase shift, a flow angle, and an acquisition time interval used to determine the Doppler phase shift. In general, the lower boundary of the detectable Doppler phase shift is limited by the phase stability of the system, while the upper boundary is limited by the π phase ambiguity. For a given range of detectable Doppler phase shift, shortening the acquisition duration will increase not only the maximum detectable velocity but unfortunately also the minimum detectable velocity, which may lead to the invisibility of a slow flow. In this paper, we present an alternative acquisition scheme for PR-DOCT that extends the lower limit of the velocity dynamic range, while maintaining the maximum detectable velocity, hence increasing the overall VDR of PR-DOCT system. The essence of the approach is to implement a technique of multi-scale measurement to simultaneously acquire multiple VDRs in a single measurement. We demonstrate an example of implementation of the technique in a dual VDR DOCT, where two Doppler maps having different detectable VDRs were simultaneously detected, processed, and displayed in real time. One was a fixed VDR DOCT capable of measuring axial velocity of up to 10.9 mm/s without phase unwrapping. The other was a variable VDR DOCT capable of adjusting its detectable VDR to reveal slow flow information down to 11.3 μm/s. The technique is shown to effectively extend the overall detectable VDR of the PR-DOCT system. Examples of real time Doppler imaging of an African frog tadpole are demonstrated using the dual-VDR DOCT system

Development Of Optical Coherence Tomography For Tissue Diagnostics

Microvasculature can be found in almost every part of the human body, including the internal organs. Importantly, abnormal changes in microvasculature are usually related to pathological development of the tissue cells. Monitoring of changes in blood flow properties in microvasculature, therefore, provides useful diagnostic information about pathological conditions in biological tissues as exemplified in glaucoma, diabetes, age related macular degeneration, port wine stains, burn-depth, and potentially skin cancer. However, the capillary network is typically only one cell in wall thickness with 5 to 10 microns in diameter and located in the dermis region of skin. Therefore, a non-invasive flow imaging technique that is capable of depth sectioning at high resolution and high speed is demanded. Optical coherence tomography (OCT), particularly after its advancement in frequency domain OCT (FD-OCT), is a promising tool for non-invasive high speed, high resolution, and high sensitivity depth-resolved imaging of biological tissues. Over the last ten years, numerous efforts have been paid to develop OCTbased flow imaging techniques. An important effort is the development of phase-resolved Doppler OCT (PR-DOCT). Phase-resolved Doppler imaging using FD-OCT is particularly of interest because of the direct access to the phase information of the depth profile signal. Furthermore, the high speed capability of FD-OCT is promising for real time flow monitoring as well as 3D flow segmentation applications. However, several challenges need to be addressed; 1) Flow in biological samples exhibits a wide dynamic range of flow velocity caused by, for example, the iv variation in the flow angles, flow diameters, and functionalities. However, the improvement in imaging speed of FD-OCT comes at the expense of a reduction in sensitivity to slow flow information and hence a reduction in detectable velocity range; 2) A structural ambiguity socalled \u27mirror image\u27 in FD-OCT prohibits the use of maximum sensitivity and imaging depth range; 3) The requirement of high lateral resolution to resolve capillary vessels requires the use of an imaging optics with high numerical aperture (NA) that leads to a reduction in depth of focus (DOF) and hence the imaging depth range (i.e. less than 100 microns) unless dynamic focusing is performed. Nevertheless, intrinsic to the mechanism of FD-OCT, dynamic focusing is not possible. In this dissertation, the implementation of PR-DOCT in a high speed swept-source based FD-OCT is investigated and optimized. An acquisition scheme as well as a processing algorithm that effectively extends the detectable velocity dynamic range of the PR-DOCT is presented. The proposed technique increased the overall detectable velocity dynamic range of PR-DOCT by about five times of that achieved by the conventional method. Furthermore, a novel technique of mirror image removal called ‘Dual-Detection FD-OCT’ (DD-FD-OCT) is presented. One of the advantages of DD-FD-OCT to Doppler imaging is that the full-range signal is achieved without manipulation of the phase relation between consecutive axial lines. Hence the full-range DD-FDOCT is fully applicable to phase-resolved Doppler detection without a reduction in detectable velocity dynamic range as normally encountered in other full-range techniques. In addition, PRDOCT can utilize the maximum SNR provided by the full-range capability. This capability is particularly useful for imaging of blood flow that locates deep below the sample surface, such as v blood flow at deep posterior human eye and blood vessels network in the dermis region of human skin. Beside high speed and functional imaging capability, another key parameter that will open path for optical diagnostics using OCT technology is high resolution imaging (i.e. in a regime of a few microns or sub-micron). Even though the lateral resolution of OCT can be independently improved by opening the NA of the imaging optics, the high lateral resolution is maintained only over a short range as limited by the depth of focus that varies inversely and quadratically with NA. Recently developed by our group, ‘Gabor-Domain Optical Coherence Microscopy’ (GD-OCM) is a novel imaging technique capable for invariant resolution of about 2-3 m over a 2 mm cubic field-of-view. This dissertation details the imaging protocol as well as the automatic data fusion method of GD-OCM developed to render an in-focus high-resolution image throughout the imaging depth of the sample in real time. For the application of absolute flow measurement as an example, the precise information about flow angle is required. GDOCM provides more precise interpretation of the tissue structures over a large field-of-view, which is necessary for accurate mapping of the flow structure and hence is promising for diagnostic applications particularly when combined with Doppler imaging. Potentially, the ability to perform high resolution OCT imaging inside the human body is useful for many diagnostic applications, such as providing an accurate map for biopsy, guiding surgical and other treatments, monitoring the functional state and/or the post-operative recovery process of internal organs, plaque detection in arteries, and early detection of cancers in the gastrointestinal tract. Endoscopic OCT utilizes a special miniature probe in the sample arm to vi access tubular organs inside the human body, such as the cardiovascular system, the lung, the gastrointestinal tract, the urinary tract, and the breast duct. We present an optical design of a dynamic focus endoscopic probe that is capable of about 4 to 6 m lateral resolution over a large working distance (i.e. up to 5 mm from the distal end of the probe). The dynamic focus capability allows integration of the endoscopic probe to GD-OCM imaging to achieve high resolution endoscopic tomograms. We envision the future of this developing technology as a solution to high resolution, minimally invasive, depth-resolved imaging of not only structure but also the microvasculature of in vivo biological tissues that will be useful for many clinical applications, such as dermatology, ophthalmology, endoscopy, and cardiology. The technology is also useful for animal study applications, such as the monitoring of an embryo’s heart for the development of animal models and monitoring of changes in blood circulation in response to external stimulus in small animal brains

Microfluidic characterization of cilia-driven fluid flow using optical coherence tomography-based particle tracking velocimetry

Motile cilia are cellular organelles that generate directional fluid flow across various epithelial surfaces including the embryonic node and respiratory mucosa. The proper functioning of cilia is necessary for normal embryo development and, for the respiratory system, the clearance of mucus and potentially harmful particulate matter. Here we show that optical coherence tomography (OCT) is well-suited for quantitatively characterizing the microfluidic-scale flow generated by motile cilia. Our imaging focuses on the ciliated epithelium of Xenopus tropicalis embryos, a genetically manipulable and experimentally tractable animal model of human disease. We show qualitative flow profile characterization using OCT-based particle pathline imaging. We show quantitative, two-dimensional, two-component flow velocity field characterization using OCT-based particle tracking velocimetry. Quantitative imaging and phenotyping of cilia-driven fluid flow using OCT will enable more detailed research in ciliary biology and in respiratory medicine

Measuring Collagen Arrangement and Its Relationship with Preterm Birth using Mueller Matrix Polarimetry

Preterm birth (PTB) is defined as delivery prior to 37 weeks of gestation. It is the leading cause of infant death worldwide, responsible for infant neurological disorders, long-term cognitive impairment, as well as chronic health issues involving the auditory, visual, digestive, and respiratory systems. In expectant mothers, causes for PTB can include infection, inflammation, vascular disease, short intervals between pregnancies, multiple gestations and genetic factors. In the U.S., PTB occurs in over 11% of births and at an elevated 18.1% in Miami-Dade County, FL; while in the developing world the incidence of PB is over 15%. Early identification of at-risk pregnancies is important for the success of medical intervention. Current diagnosis methodologies of PTB include ultrasound imaging of cervical length and fetal fibronectin assay but have low positive predictive power. Compared to the markers targeted by current diagnosis methodologies, collagen content in the cervix changes more drastically throughout the course of gestation due to its link to changes in load bearing capacity that occur during the phases of pregnancy. Mueller matrix polarimetry is capable of characterizing changes in collagen without making contact with patients and may prove to be an improvement to current diagnosis methodologies. A clear difference is seen in collagen orientation between nonpregnant and pregnant patients. The development of a new imaging modality aimed at assessing early changes in collagen arrangement in the cervix may improve risk determination of PTB and reduce the morbidity of the condition. Earlier prediction of PTB could improve outcomes by allowing longer intervention times to prolong gestation time for the infant in the womb. A more reliable quantitative predictor may also lead to development of more treatment options

Photoacoustic Reporter Gene Imaging And Optical Coherence Computed Tomography

Advances in imaging technologies have always been the major driving forces for the evolution of biomedical research. Compared with other modalities, optical imaging possesses several prominent merits. Because light interacts with tissue at the microscopic level through many distinct physical mechanisms, optical methods allow sensitive exploration of various aspects of the life down to the single-molecule level. From the technical perspective, optical systems utilize safe non-ionizing radiation, could be implemented at relatively low cost, also have the potential to be miniaturized for portable or endoscopic applications. As a result, optical imaging tools are playing an increasingly important role in both laboratorial research and clinical practice. Among them, photoacoustic imaging: PAI) and optical coherence tomography: OCT) are the two fastest growing branches. PAI measures the laser-induced acoustic wave, and produces high-resolution images of the optically absorbing features of tissue at multiple length-scales. OCT detects singly backscattered photons, and enables real-time high-resolution in vivo biopsy of tissue up to an optical transport mean-free-path. My doctoral research is focused on developing three novel optical imaging techniques based on the spirits of PAI and OCT. In the first part of this study, we established a new paradigm to visualize gene expression in vivo based on optical absorption. In the post-genomic era, we are now being challenged to develop novel molecular imaging methods to identify the functions of genes. PAI can detect specific molecules according to their characteristic absorption spectra, thus is a promising candidate for molecular imaging of gene expression. The full potential of photoacoustic molecular imaging still remains to be explored. For the first time, we demonstrated imaging gene expression by PAI in living mice and rats, using a chromogenic lacZ/X-gal reporter gene system. We demonstrated the expression of the lacZ reporter gene can be detected by PAI as deep as 5 cm inside tissue. In addition, we showcased that PAI could follow gene expression from the microscopic to the macroscopic level. This work represents one of the pioneering efforts to extend photoacoustic methods for molecular imaging. In the second part of this study, we developed a novel multimodal microscope, called the integrated photoacoustic and optical coherence microscope: iPOM), which combines PAI and OCT in a single imaging platform. PAI is predominantly sensitive to optical absorption, while OCT exploits optical scattering. By combining their naturally complementary imaging contrasts, iPOM can provide comprehensive information about biological tissue. We designed and built a reflection-mode prototype of iPOM, which fuses optical-resolution photoacoustic microscopy with spectral-domain optical coherence tomography. The potential applications of iPOM in studying cutaneous and ocular microcirculation, and tissue engineering were demonstrated. Finally, we invented a new optical tomography, named optical coherence computed tomography: optical CCT), which overcomes several major limitations of OCT. OCT relies on singly backscattered photons to obtain high-resolution images. Its image quality degrades fast with the increase of the depth, because the multiply scattered photons quickly become dominant at a penetration larger than 500 &mum. As a result, OCT can only effectively penetrate ~1 mm into highly scattering tissue like skin. In addition, OCT is mainly sensitive to optical scattering, which does not reflect the molecular content of tissue directly. Optical CCT measures both singly and multiply scattered photons using a low-coherence interferometer. We make use of both types of photons by adopting a model-based reconstruction algorithm. The light-tissue interaction model was established using the time-resolved Monte Carlo method. The optical properties of the tissue were reconstructed from measurements by solving the inverse radiative transport problem under the first Born approximation. As a result, optical CCT could image deeper than OCT, and provide extra molecule-specific contrasts, such as optical absorption. We designed and built the first optical CCT system. In experiments, absorbing inclusions of 100 &mum diameter were imaged with consistent quality through a 2.6-mm-thick: equivalent to ~3 transport mean-free-paths) tissue-mimicking phantom