血管系統的功能與如何去維持細胞的恆定狀態有關，而利用超音波來量測血管動態功能如血流速度則是最常被用來評估血管系統的功能。目前超音波在量測血管動態功能上主要藉由超音波都卜勒(Doppler)技術，包含了能量都卜勒、彩色都卜勒以及脈衝波都卜勒量測方式。然而，一般的都卜勒技術因為受限於都卜勒角度的問題，沒辦法去量測複雜移動情況(如擾流或是渦流)，儘管有許多超音波向量流的技術被提出來解決此項問題，目前向量流技術的時間以及空間解析度不足以來清楚解析複雜移動方式，特別是針對在臨床前的小動物研究上，向量流的技術仍然有許多限制。
有鑒於此，本論文目的為發展高解析度(包含時間及空間)的向量都卜勒影像(high-resolution vector Doppler imaging, HRVDI)來準確的評估複雜的血流流動或是物體移動模型，並且能應用於評估血管功能。為了實現此目標，超音波影像擷取部分使用了高頻(40 MHz)超音波探頭並搭配超快速的平面波發射成像。同時，利用多角度的平面波發射來評估向量流並能準確評估每個立體位置點(二維空間與一維時間)的向量速度。此論文中闡述演算法的驗證及實際用於小動物研究中有關血管功能的研究，並延伸至在人體上的研究。
有關HRVDI的演算法的建立及相關驗證，HRVDI主要利用不同平面波發射角度向所得到的彩色都卜勒結果作為基礎，對於單一角度發射的彩色都卜勒來說，僅能量測到平行於波前進方向的速度，實際量測到準確流速需藉由使用者定義都卜勒角度，而在多重發射下，可獲得在不同發射方向的速度，並可藉由多重波束方式在不定義都卜勒角度情況下得到準確流速及方向。HRVDI首先被用來評估血管動態功能中有關腦血管的的血流動力學，特別是針對皮質層的穿透及上升小血管，以往因為時空間解析度不足所以難以量測到此區域的變化。首先藉由奇異值濾波器來偵測血流(特別針對小血管)，接著應用以形態學為基礎的Bowler-hat轉換以及海森血管增強技術(Hessian-based vessel enhancement)來抑制雜訊並改善血管的可視性；同時，以奇異值為基礎的自適應性濾波器也被提出來改善小血管流速的偵測。驗證部分則使用微流管來完成，誤差落於10 %以內。整合這些技術，高時空解析度的向量微都卜勒影像(high-spatiotemporal-resolution vector micro-Doppler imaging, HVμDI)被提出來視覺化及量化腦血管動力情形，最小可觀測的血管尺寸為38 μm而有效的影像解析度為500 Hz。實驗部分則在野生以及阿茲海默症的小鼠腦上執行。腦血管直徑、血流速度、血管曲折度、密度以及脈動性被用來做為評估參數。除此之外，HVμDI更被用來檢驗不同順時下血流的方向性。所有結果均顯示阿茲海默症造成的病變可被HVμDI診斷。
在第二部分，HRVDI被進一步用來探討在小鼠的心血管系統中有關冠狀動脈的血管動態功能。當冠狀動脈呈現粥狀硬化情形，會導致心肌供氧量不足並使心肌產生硬化，因此可藉由評估心肌的形變影像來評估冠狀動脈的血管動態功能。HRVDI可以提供每個立體像素點的向量速度，故很適合用來做高解析度心肌形變影像。一個自動兩階段隨機迭代方式被提出來量測心肌形變，驗證部分則是藉由量測氣球仿體在充氣及洩氣時管壁厚度的變化來完成。仿體結果部分顯示了高的相關係數(r=0.84)。體內研究則使用野生(wild-type, WT)及心肌梗塞(myocardial infarction, MI)的老鼠。結果顯示MI老鼠在逕向形變(長軸及短軸)、環狀形變、及橫向形變均有明顯下降，且此趨勢能藉由所提出的心肌影像來清楚觀測及評估，包含了不同局部區域上的病變情形。另一方面也能清楚量測出不同心肌層的形變數值[WT 老鼠: −22.0 pm 1.2 % (內層) 及 −16.8 pm 0.9 % (外層)]；在MI老鼠上分層形變則沒有明顯差異。此實驗很好的展示了藉由HRVDI可用來做出動態的高解析度影像並能應用於心臟學研究上。
第三部分則是將HRVDI及其衍伸的心肌形變影像應用於斑馬魚的心臟再生研究。斑馬魚心臟尺寸較小鼠心臟小，因此更適合所提出的高解析度超音波向量都卜勒影像。斑馬魚的心肌構造可分為兩部分，外層為完整的心肌構造，內層則為有許多層狀小樑(trabeculae)所組成的複雜構造，因此難以直接藉由影像辨別出心肌位置。因此，在班馬魚心臟研究部分首先利用奇異值濾波器來偵測心臟血流區域，再將此偵測出的區域從心臟大致位置中去除來識別心肌區域，此識別出的區域則被用來評估心肌的速度以及型變。驗證方面利用自製的不同形狀的流管來進行驗證，血管偵測的誤差則約為6 %左右。實驗方面使用AB-line的成年斑馬魚，並利用心肌凍損手術來評估心臟再生。藉由所提出的技術可直接視覺化在受傷前後以及恢復中心肌速度及型變的變化，心肌動損手術後，在晚期舒張時期的心肌速度有顯著的下降(約−2.1 mm/s)，而形變則在受傷區域有顯著的差異(約−4.4 %)。在術後第14天，速度及型變接回覆正常標準值。此部分展示了所提出的技術可運用於小動物斑馬魚的實驗，並能用來評估在心臟再生期間的血管動態功能。
在最後一部分則將HRVDI運用於人體上的量測，在人體量測上血流速度較快，在使用高頻超音波的情況下較容易超過可量測的速度範圍，並產生相位混疊的情形，而當相位混疊發生時，速度的誤差會大幅增加，甚至超過100 %，故相位展開基於時間及空間的連續性被用來解決此問題。直管仿體以及窄化管仿體被用來驗證相位展開的演算法，在流速高於可量測範圍的情況下，藉由相位展開演算法後大部分流速誤差在10 %以內。HRVDI的可行性在人體的靜脈血流上做初步測試。HRVDI被用來量測靜脈瓣附近不同位置的速度，同時也測試了不同姿勢下的流速情形。結果顯示流速在靜脈瓣間最快且在坐姿姿勢時流速會大幅下降。HRVDI並能用來動態描繪靜脈瓣間高速流噴射的情形以及在靜脈瓣竇所造成的低流速渦流情形。因此，HRVDI在人體研究的可行性被驗證以及展示。
在此論文中，基於都卜勒技術的HRVDI被提出，並藉由超快速的高頻超音波系統來實現。此技術能夠提出在影像範圍內每個立體像素點的向量速度，以此技術為基礎更能被用來動態視覺化及量化腦血管及心血管的動態功能，並延伸至人體靜脈血管功能的研究。所有結果均顯示高解析度的向量都卜勒影像可藉由超快速高頻超音波影像來實現並能實際應在多種研究上(包含小動物研究及人體研究)來評估血管功能。

The function of the vascular systems such as the cardiovascular or cerebral vascular systems is associated with the maintenance of the cellular homeostasis. In order to investigate the function of the vascular system, the golden standard is to use the ultrasound for measuring the dynamic vascular function such as the vascular velocity. Currently, measuring the dynamic vascular function through ultrasound is typically performed using ultrasound Doppler flow measurements (including power Doppler, color Doppler and pulsed-wave Doppler measurements). However, conventional Doppler measurements are prone to error due to the inherent Doppler angle issue in the complex turbulent or vortex flow region. Although several ultrasound vector flow imaging techniques have been developed. The spatial and temporal resolutions of current vector flow techniques are insufficient to depict the complex flow clearly, especially for the preclinical small-animal imaging.
Therefore, the goal of the present thesis aims to develop a high-resolution (both temporal and spatial) vector Doppler imaging (HRVDI) for the accurately estimating the complex flow (or motion) pattern and assessing the dynamic vascular function. To achieve this goal, a 40-MHz high-frequency linear-array transducer with ultrafast plane-wave transmitting is utilized for the data acquisition. Multi-angle plane-wave transmitting is utilized for estimation the vector flow imaging, which ensure the capability of the vector information in every voxel (two-dimensional spatial and one-dimensional temporal). Validation of the algorithms and the assessing vascular function in both small animals and humans to confirm the feasibility and capability in this thesis.
First part illustrates the algorithm of HRVDI and the phantom validation. The algorithm of HRVDI is based on the color Doppler measurements from different plane-wave transmitting angles. For color Doppler measurement from single plane-wave transmitting angles, only the velocity along the beam direction can be measured and a Doppler angle is needed to be manually determined for accurate measurement; while HRVDI utilized the color Doppler results from different transmitting angles and the accurate velocity (magnitude and direction) can be evaluated through multi-beam strategy. HRVDI is firstly applied for the dynamic vascular function related to cerebrovascular hemodynamics, especially for the small cortical penetrating and ascending vessels, which remains challenging in current ultrasound modalities due to the insufficient spatiotemporal resolution. Singular value decomposition (SVD) filtering is firstly utilized to extract the blood flow signals (especially for the small vessels). Then, morphology-based methods of bowler-hat transform and Hessian-based vessel enhancement are applied to suppress the noise and improve the visibility of the small vessels. Also, an automatically adaptively filtering strategy based on SVD operation is proposed for accurately measurement in small vessels. A microtube flow phantom is established for the validation and the errors are < 10 %. With these processing, a high-spatiotemporal-resolution vector micro-Doppler imaging (HVμDI) is developed for visualizing and quantifying cerebrovascular hemodynamics. The minimal detectable vascular size is 38 μm and the temporal resolution is 500 Hz. Experiments are carried out using WT and Alzheimer’s disease (AD) mice. Cerebrovascular vessel diameters, velocity, tortuosity, density and is utilized for quantification. In addition, HVμDI ensures the capability of assessing the instant flow directionality. Results shows the alteration due to AD can be assessed through HVμDI.
HRVDI is further applied for evaluating dynamic vascular function of the coronary atherosclerosis in the mice cardiovascular system. Coronary atherosclerosis disease causes insufficient blood supple to the heart muscle, leading to myocardial stiffness. Myocardial strain imaging can be utilized for the assessment of stiffness and thereby can reflect the disorder in coronary disease. HRVDI can be utilized for a high-resolution myocardial strain imaging since the vector velocity in every voxel can be measured. An automatic two-step randomized iterative approach is proposed for the strain estimation. Validation is carried out through balloon phantom experiments by estimating the change in balloon wall thickness due to the inflation or dilation. The experiments demonstrate a high coefficient of the correlation (r=0.84). In vivo experiments are performed for wild-type (WT) and myocardial infarction (MI) mice. Strains including radial (long and short axis), longitudinal and circumferential strains decreased largely in MI mice and the decrease can be dynamically visualized through high-resolution strain imaging. Regional analysis shows a significant decrease in the ventricle apical anterior region of MI mice. Layer-based longitudinal strain curves are also measured and the peak strain values for WT mice were −22.0 pm 1.2 % and −16.8 pm 0.9 % in the endocardial (inner) and epicardial (outer) layers, respectively. However, no significant difference in the layer-based values is noted for the MI mice. The experimental results demonstrate that the proposed dynamic cardiac strain imaging through HRVDI can be useful in high-performance small-animal imaging of cardiovascular research.
Subsequently, HRVDI and the proposed strain imaging are applied for the investigation of heart regeneration in adult zebrafish. Due to its smaller size compared to the one of mice, the proposed HRVDI becomes even more suitable for zebrafish. The myocardium of zebrafish can be separated into two parts: a compact outer compact myocardium and an inner myofibers organized into trabeculae. It’s difficult to recognize the myocardial region directly through the images. Therefore, cardio flow extraction through SVD filtering is conducted and the myocardial region is recognized by subtracting the extracted cardio flow region from the segmented ventricular position, and the recognized region is utilized for the evaluation of myocardial velocity and strain. Validation is performed through in-house manufactured flow phantom and the error of the flow extraction is around 6 %. In vivo experiments are carried out using AB-line adult zebrafish and cryoinjury is induced for evaluating heart regeneration. With the proposed technique, the difference in myocardial velocity and strain during heart regeneration is clearly visualized and quantified. After cryoinjury, myocardial velocity significantly decreases (around -2.1 mm/s); while myocardial strain significantly decreases (around -4.4 %) in the injured region. Both the myocardial velocity and strain returns to the baseline value at day 14 after cryoinjury. The experimental results demonstrate that the proposed technique can be implemented on the small-animal zebrafish experiment and utilized for the investigation of heart regeneration.
Finally, HRVDI is utilized for measurement in human study. Since the flow velocity is faster in human than in small animals, the flow velocity may exceed the maximal measured velocity due to the use of high-frequency ultrasound, leading to aliasing. When aliasing occurs, the velocity estimation becomes reliable (error may even exceed 100 %). Therefore, phase-unwrapping processing based on the spatial and temporal continuity is performed to avoid the aliasing. With phase-unwrapping processing, most of errors are less than 10 %. The preliminary feasibility of HRVDI in human study are demonstrated through human venous flow, which is associated with the chronic venous disease. Flow velocities at different location in sitting and standing positions are evaluated. The velocity is faster between the leaflets and the velocity decreases in sitting position. HRVDI is able to dynamically depict the high-speed jet phenomena between the venous valve leaflets and low-speed vortex phenomena in the sinus pocket. Hence, the capability and feasibility of HRVDI in human study is proved and demonstrated.
In this thesis, HRVDI are proposed and implemented on various dynamic vascular functions by using ultrafast high-frequency ultrasound system. This imaging methodology is able to provide the vector flow information in every voxel in all the field of view. The dynamic vascular function, including the cerebral hemodynamics, myocardial strain imaging and the venous vortex flow, can be dynamically visualized and quantified. All results reveal that the proposed HRVDI can achieved through ultrafast high-frequency imaging and can be applied on various study (from small animal to human) to evaluate the dynamic vascular function.

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