光學(xué)斷層分子成像定量分析及其應(yīng)用
發(fā)布時(shí)間：2018-08-23 21:44
【摘要】：近十年來(lái)，光學(xué)斷層分子成像（Optical Molecular Tomography, OMT）因其在預(yù)臨床的重大疾病研究應(yīng)用中所表現(xiàn)出的良好性能，受到了學(xué)者們的廣泛關(guān)注。光學(xué)斷層分子成像由于具有靈敏度高、成本相對(duì)低廉、可能實(shí)現(xiàn)絕對(duì)定量等諸多優(yōu)點(diǎn)，因而在預(yù)臨床和臨床的惡性腫瘤、心血管疾病、神經(jīng)系統(tǒng)病變等重大疾病研究中得到了廣泛的應(yīng)用。作為光學(xué)斷層分子成像的典型代表，生物發(fā)光斷層成像及超高分辨率micro-CT混合成像系統(tǒng)以其高靈敏性、高特異性及高分辨的解剖結(jié)構(gòu)等優(yōu)點(diǎn)，在生物醫(yī)學(xué)研究中具有廣闊的應(yīng)用前景。本文對(duì)光學(xué)斷層分子成像的絕對(duì)定量分析方法進(jìn)行了深入研究，并就生物發(fā)光斷層分子成像和超高分辨率micro-CT混合成像系統(tǒng)對(duì)胃癌和干細(xì)胞移植治療缺血性疾病的生物醫(yī)學(xué)應(yīng)用進(jìn)行了系統(tǒng)的成像研究，本論文的主要研究?jī)?nèi)容包括： 1.構(gòu)建了生物發(fā)光斷層分子成像和超高分辨率micro-CT混合成像系統(tǒng)，并對(duì)生物發(fā)光斷層分子成像系統(tǒng)進(jìn)行了定量校準(zhǔn)。針對(duì)預(yù)臨床小動(dòng)物研究的需要，，構(gòu)建了生物發(fā)光斷層分子成像系統(tǒng)，并對(duì)成像系統(tǒng)CCD相機(jī)進(jìn)行了像元響應(yīng)的非均勻性校準(zhǔn)和絕對(duì)強(qiáng)度的定量校準(zhǔn)。針對(duì)光學(xué)斷層分子成像空間位置的差異構(gòu)建了視場(chǎng)函數(shù)，對(duì)進(jìn)入光學(xué)成像系統(tǒng)的光通量進(jìn)行了定量分析。構(gòu)建了超高分辨率micro-CT成像系統(tǒng)，為本論文的下肢缺血模型的血管新生定量研究奠定了基礎(chǔ)。 2.對(duì)生物發(fā)光斷層分子成像定量重建方法進(jìn)行了研究。基于有限元網(wǎng)格剖分方法，提出了總能量定量重建的策略。通過(guò)已知光源移植到小動(dòng)物體內(nèi)，驗(yàn)證了總能量重建的可行性。建立了基于報(bào)告基因系統(tǒng)的光學(xué)探針?biāo)从车纳飳W(xué)參數(shù)與成像系統(tǒng)中總能量重建的物理參數(shù)之間的內(nèi)在聯(lián)系，實(shí)現(xiàn)了生物發(fā)光斷層分子成像系統(tǒng)對(duì)腫瘤細(xì)胞和干細(xì)胞的細(xì)胞數(shù)量的定量分析。通過(guò)PC-3M-luc-C6前列腺癌和A549-luc-C8非小細(xì)胞肺癌細(xì)胞系以及脂肪來(lái)源的間充質(zhì)干細(xì)胞的在體定量研究，證實(shí)了細(xì)胞水平定量分析的可行性和適用性。 3.研究了光學(xué)斷層分子成像在光學(xué)探針?lè)逯挡ㄩL(zhǎng)偏移及解剖結(jié)構(gòu)模型偏差等情況下引入的重建誤差及其貢獻(xiàn)率的大小。以生物發(fā)光斷層成像為例分析了光學(xué)斷層分子成像的重建誤差的主要來(lái)源。（1）在假定光學(xué)參數(shù)準(zhǔn)確的前提下，研究了動(dòng)物模型解剖結(jié)構(gòu)偏差帶來(lái)的光學(xué)重建在定位和定量上的偏差。（2）在假定解剖結(jié)構(gòu)準(zhǔn)確的情況下，研究了光學(xué)探針?lè)逯挡ㄩL(zhǎng)偏移對(duì)重建誤差在定位和定量方面的影響。通過(guò)對(duì)解剖結(jié)構(gòu)偏差以及峰值波長(zhǎng)偏移的實(shí)驗(yàn)數(shù)據(jù)的系統(tǒng)測(cè)試、重建及數(shù)據(jù)分析，表明二者均會(huì)對(duì)重建的定位和定量帶來(lái)較大的影響；其中，當(dāng)光學(xué)探針?lè)逯挡ㄩL(zhǎng)在640nm以下時(shí)，解剖結(jié)構(gòu)的偏差對(duì)重建在定位和定量有更大的影響。實(shí)驗(yàn)數(shù)據(jù)也揭示了光學(xué)探針?lè)逯挡ㄩL(zhǎng)漂移量在±10nm范圍內(nèi)、動(dòng)物模型的解剖結(jié)構(gòu)偏差在亞毫米級(jí)別以內(nèi)時(shí)都可以得到較理想的定位和定量重建結(jié)果。 4.利用自主研發(fā)的混合成像系統(tǒng)對(duì)胃癌的生物學(xué)行為進(jìn)行定量成像研究。利用生物發(fā)光斷層分子成像對(duì)胃癌原位動(dòng)物模型實(shí)現(xiàn)在體定量分析以及腫瘤模型的長(zhǎng)程動(dòng)態(tài)觀測(cè)，為胃癌的預(yù)臨床研究提供了新的成像方法和成像工具。通過(guò)micro-CT造影成像精確計(jì)算出腫瘤的實(shí)際體積大小，同時(shí)在研究中提出了新穎的低成本歐乃派克時(shí)間累積對(duì)比增強(qiáng)造影方法，可以實(shí)現(xiàn)微小胃癌轉(zhuǎn)移灶的在體定位和定量分析。 5.對(duì)干細(xì)胞移植治療缺血性疾病促進(jìn)血管新生開(kāi)展了混合成像系統(tǒng)的定量示蹤和基于空間血管體積因數(shù)SVVF定量評(píng)估的成像研究。（1）采用生物發(fā)光斷層分子成像及超高分辨率micro-CT混合成像系統(tǒng)在三維空間實(shí)現(xiàn)干細(xì)胞的在體定量示蹤以及血管新生的血管灌注分析，實(shí)現(xiàn)對(duì)干細(xì)胞在體空間分布和細(xì)胞數(shù)量的定量示蹤；并根據(jù)干細(xì)胞的遷移、存活數(shù)量和空間分布，進(jìn)一步交互式實(shí)施高分辨的微血管網(wǎng)絡(luò)密度的定量評(píng)估。（2）提出了空間血管體積因數(shù)（Spatial VascularVolume Fraction, SVVF）的概念，并將其用于小鼠下肢缺血模型血管新生的定量評(píng)估。（3）通過(guò)血管體積因數(shù)SVVF的系列成像研究發(fā)現(xiàn)在時(shí)間和空間上血管新生的量化指標(biāo)，進(jìn)而探討性地分析了干細(xì)胞移植后旁分泌的作用效果和作用半徑。（4）通過(guò)多種傳統(tǒng)的方法對(duì)干細(xì)胞移植后的血管新生進(jìn)行了交叉驗(yàn)證。利用血管鑄型驗(yàn)證血管新生的宏觀變化情況；通過(guò)掃描電鏡成像證實(shí)了在干細(xì)胞移植后促進(jìn)血管新生作用的不同時(shí)間點(diǎn)血管新生的微觀結(jié)構(gòu)及發(fā)芽情況；通過(guò)共聚焦熒光顯微成像證實(shí)了eGFP轉(zhuǎn)基因小鼠來(lái)源的脂肪間充質(zhì)干細(xì)胞在缺血組織的存在性和血管新生的情況；通過(guò)激光多普勒成像觀測(cè)血流灌注的恢復(fù)情況。本文提出的SVVF方法以及交叉驗(yàn)證的結(jié)果一致發(fā)現(xiàn)，脂肪來(lái)源的間充質(zhì)干細(xì)胞移植與對(duì)照組相比可以明顯地增大了血管密度，進(jìn)而證實(shí)脂肪來(lái)源的間充質(zhì)干細(xì)胞移植促進(jìn)了血管新生。綜上，在三維空間的血管體積因數(shù)成像以干細(xì)胞在三維空間的在體示蹤存活的數(shù)量和空間分布作為反饋信息，進(jìn)而對(duì)血管新生進(jìn)行定量成像研究。生物發(fā)光斷層成像及超高分辨率micro-CT混合成像系統(tǒng)在三維空間為基于細(xì)胞移植治療的機(jī)制研究和長(zhǎng)程監(jiān)測(cè)研究提供了新穎的成像工具和功能評(píng)估手段。 本論文的研究涉及到的動(dòng)物實(shí)驗(yàn)均按照動(dòng)物操作規(guī)程執(zhí)行，并得到了本單位實(shí)驗(yàn)動(dòng)物保護(hù)委員會(huì)的批準(zhǔn)。
[Abstract]:Over the past decade, optical molecular tomography (OMT) has attracted considerable attention due to its excellent performance in the research and application of preclinical major diseases. Bioluminescence tomography and ultra-high resolution micro-CT hybrid imaging system have been widely used in the study of preclinical and clinical malignancies, cardiovascular diseases, neuropathy and other major diseases. Absolute quantitative analysis methods of optical tomography molecular imaging are studied in this paper. Biomedical applications of bioluminescent tomography molecular imaging and ultra-high resolution micro-CT hybrid imaging system for gastric cancer and stem cell transplantation in the treatment of ischemic diseases are systematically studied. In imaging research, the main contents of this thesis include:
1. Bioluminescent tomography molecular imaging system and ultra-high resolution micro-CT hybrid imaging system were constructed, and the bioluminescent tomography molecular imaging system was calibrated quantitatively. Calibration and quantitative calibration of absolute intensity.The field-of-view function is constructed to quantify the luminous flux entering the optical imaging system according to the spatial position difference of optical tomography molecular imaging.The ultra-high resolution micro-CT imaging system is constructed,which lays a foundation for the quantitative study of angiogenesis in the lower limb ischemia model in this paper.
2. Quantitative reconstruction method of bioluminescent tomography molecular imaging is studied. Based on finite element mesh generation method, a strategy of total energy reconstruction is proposed. The feasibility of total energy reconstruction is verified by transplanting known light sources into small animals. The biological parameters reflected by optical probes based on reporter gene system are established. The in vivo quantitative analysis of tumor cells and stem cells by bioluminescent tomography molecular imaging system has been achieved through the intrinsic relationship with the physical parameters of total energy reconstruction in the imaging system. PC-3M-luc-C6 prostate cancer and A549-luc-C8 non-small cell lung cancer cell lines and adipose-derived mesenchymal stem cells were quantified in vivo. The study confirmed the feasibility and applicability of quantitative analysis of cell level.
3. The reconstruction error and its contribution rate of optical tomography molecular imaging under the condition of peak wavelength shift of optical probe and deviation of anatomical structure model are studied. The main source of reconstruction error of optical tomography molecular imaging is analyzed with bioluminescence tomography as an example. (1) On the premise of accurate optical parameters, the reconstruction error is studied. (2) Under the assumption of accurate anatomical structure, the effects of peak wavelength offset of optical probe on the location and quantification of optical reconstruction errors were studied. The results of reconstruction and data analysis show that both of them have a great influence on the localization and quantification of the reconstructed image. When the peak wavelength of the optical probe is below 640 nm, the deviation of the anatomical structure has a greater influence on the localization and quantification of the reconstructed image. Satisfactory localization and quantitative reconstruction results can be obtained when the anatomical structure deviation is within the sub-millimeter level.
4. Quantitative imaging of biological behavior of gastric cancer was studied by using a self-developed hybrid imaging system. Quantitative analysis of animal models of gastric cancer in situ and long-term dynamic observation of tumor models were carried out by using bioluminescent tomography, which provided a new imaging method and imaging tool for preclinical research of gastric cancer. Ro-CT imaging can accurately calculate the actual size of the tumor, and a novel and low-cost Ohnipec time-cumulative contrast-enhanced imaging method is proposed to localize and quantify the metastasis of gastric cancer in vivo.
5. Quantitative tracing of stem cell transplantation for angiogenesis in ischemic diseases and quantitative evaluation of SVVF based on spatial vascular volume factor were carried out. (1) In vivo quantitative identification of stem cells in three-dimensional space was achieved by using bioluminescent tomography and ultra-high resolution micro-CT hybrid imaging system. Tracing and angiogenesis perfusion analysis were used to quantify the spatial distribution and cell number of stem cells in vivo, and high-resolution quantitative evaluation of microvascular network density was carried out interactively according to the migration, survival and spatial distribution of stem cells. (2) Spatial Vascular Volume was proposed. The concept of e-Fraction (SVVF) was applied to quantitatively evaluate angiogenesis in a mouse model of lower limb ischemia. (3) Quantitative parameters of angiogenesis were found in time and space by serial imaging of SVVF, and the paracrine effect and radius of action after stem cell transplantation were discussed. Angiogenesis after stem cell transplantation was cross-validated by traditional methods. The macroscopic changes of angiogenesis were verified by vascular casting; the microscopic structure and germination of angiogenesis at different time points after stem cell transplantation were confirmed by scanning electron microscopy; and the confocal fluorescence was used. Microscopic imaging confirmed the existence and angiogenesis of adipose-derived mesenchymal stem cells from eGFP transgenic mice in ischemic tissues, and the recovery of blood perfusion was observed by laser Doppler imaging. In conclusion, the number and spatial distribution of stem cells traced in vivo in three-dimensional space are used as feedback information in three-dimensional space to quantify angiogenesis. Bioluminescent tomography and ultra-high resolution micro-CT hybrid imaging system provide novel imaging tools and functional evaluation tools for mechanism research and long-term monitoring research based on cell transplantation therapy in three-dimensional space.
The animal experiments involved in this paper are carried out according to the animal operation rules, and have been approved by the Laboratory Animal Protection Committee of this unit.
【學(xué)位授予單位】：西安電子科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R318.51;R735.2
本文編號(hào)：2200012
[Abstract]:Over the past decade, optical molecular tomography (OMT) has attracted considerable attention due to its excellent performance in the research and application of preclinical major diseases. Bioluminescence tomography and ultra-high resolution micro-CT hybrid imaging system have been widely used in the study of preclinical and clinical malignancies, cardiovascular diseases, neuropathy and other major diseases. Absolute quantitative analysis methods of optical tomography molecular imaging are studied in this paper. Biomedical applications of bioluminescent tomography molecular imaging and ultra-high resolution micro-CT hybrid imaging system for gastric cancer and stem cell transplantation in the treatment of ischemic diseases are systematically studied. In imaging research, the main contents of this thesis include:
1. Bioluminescent tomography molecular imaging system and ultra-high resolution micro-CT hybrid imaging system were constructed, and the bioluminescent tomography molecular imaging system was calibrated quantitatively. Calibration and quantitative calibration of absolute intensity.The field-of-view function is constructed to quantify the luminous flux entering the optical imaging system according to the spatial position difference of optical tomography molecular imaging.The ultra-high resolution micro-CT imaging system is constructed,which lays a foundation for the quantitative study of angiogenesis in the lower limb ischemia model in this paper.
2. Quantitative reconstruction method of bioluminescent tomography molecular imaging is studied. Based on finite element mesh generation method, a strategy of total energy reconstruction is proposed. The feasibility of total energy reconstruction is verified by transplanting known light sources into small animals. The biological parameters reflected by optical probes based on reporter gene system are established. The in vivo quantitative analysis of tumor cells and stem cells by bioluminescent tomography molecular imaging system has been achieved through the intrinsic relationship with the physical parameters of total energy reconstruction in the imaging system. PC-3M-luc-C6 prostate cancer and A549-luc-C8 non-small cell lung cancer cell lines and adipose-derived mesenchymal stem cells were quantified in vivo. The study confirmed the feasibility and applicability of quantitative analysis of cell level.
3. The reconstruction error and its contribution rate of optical tomography molecular imaging under the condition of peak wavelength shift of optical probe and deviation of anatomical structure model are studied. The main source of reconstruction error of optical tomography molecular imaging is analyzed with bioluminescence tomography as an example. (1) On the premise of accurate optical parameters, the reconstruction error is studied. (2) Under the assumption of accurate anatomical structure, the effects of peak wavelength offset of optical probe on the location and quantification of optical reconstruction errors were studied. The results of reconstruction and data analysis show that both of them have a great influence on the localization and quantification of the reconstructed image. When the peak wavelength of the optical probe is below 640 nm, the deviation of the anatomical structure has a greater influence on the localization and quantification of the reconstructed image. Satisfactory localization and quantitative reconstruction results can be obtained when the anatomical structure deviation is within the sub-millimeter level.
4. Quantitative imaging of biological behavior of gastric cancer was studied by using a self-developed hybrid imaging system. Quantitative analysis of animal models of gastric cancer in situ and long-term dynamic observation of tumor models were carried out by using bioluminescent tomography, which provided a new imaging method and imaging tool for preclinical research of gastric cancer. Ro-CT imaging can accurately calculate the actual size of the tumor, and a novel and low-cost Ohnipec time-cumulative contrast-enhanced imaging method is proposed to localize and quantify the metastasis of gastric cancer in vivo.
5. Quantitative tracing of stem cell transplantation for angiogenesis in ischemic diseases and quantitative evaluation of SVVF based on spatial vascular volume factor were carried out. (1) In vivo quantitative identification of stem cells in three-dimensional space was achieved by using bioluminescent tomography and ultra-high resolution micro-CT hybrid imaging system. Tracing and angiogenesis perfusion analysis were used to quantify the spatial distribution and cell number of stem cells in vivo, and high-resolution quantitative evaluation of microvascular network density was carried out interactively according to the migration, survival and spatial distribution of stem cells. (2) Spatial Vascular Volume was proposed. The concept of e-Fraction (SVVF) was applied to quantitatively evaluate angiogenesis in a mouse model of lower limb ischemia. (3) Quantitative parameters of angiogenesis were found in time and space by serial imaging of SVVF, and the paracrine effect and radius of action after stem cell transplantation were discussed. Angiogenesis after stem cell transplantation was cross-validated by traditional methods. The macroscopic changes of angiogenesis were verified by vascular casting; the microscopic structure and germination of angiogenesis at different time points after stem cell transplantation were confirmed by scanning electron microscopy; and the confocal fluorescence was used. Microscopic imaging confirmed the existence and angiogenesis of adipose-derived mesenchymal stem cells from eGFP transgenic mice in ischemic tissues, and the recovery of blood perfusion was observed by laser Doppler imaging. In conclusion, the number and spatial distribution of stem cells traced in vivo in three-dimensional space are used as feedback information in three-dimensional space to quantify angiogenesis. Bioluminescent tomography and ultra-high resolution micro-CT hybrid imaging system provide novel imaging tools and functional evaluation tools for mechanism research and long-term monitoring research based on cell transplantation therapy in three-dimensional space.
The animal experiments involved in this paper are carried out according to the animal operation rules, and have been approved by the Laboratory Animal Protection Committee of this unit.
【學(xué)位授予單位】：西安電子科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R318.51;R735.2
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 ;Optical properties of human normal small intestine tissue determined by Kubelka-Munk method in vitro[J];World Journal of Gastroenterology;2003年09期
本文編號(hào)：2200012
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