臺灣大學: 電機工程學研究所孫啟光李文正Lee, Wen-JengWen-JengLee2013-03-272018-07-062013-03-272018-07-062012http://ntur.lib.ntu.edu.tw//handle/246246/253883影像醫學傳統上使用的工具包含X光、磁振造影、超音波與核子醫學四種儀器設備。這四種儀器，已經廣泛地應用在各種疾病的非侵入式診斷。然而，上述四種檢查工具的空間解析度大約可以只能分辨至毫米級的構造，對於細胞、組織構造的微小變異，無法提供有效的檢驗。 近年來新興的生醫光電(biophotonic)技術，擁有高解析度的影像，能夠分辨出微米甚至奈米級的構造。利用生物體經由高強度雷射光的照射下所自然產生的二倍頻與三倍頻訊號，進行的倍頻顯微術，能夠獲得高解析、非侵入式的虛擬光學切片。 本論文探討使用倍頻顯微術進行之虛擬光學切片術的臨床醫學應用價值。論文中針對三種臨床應用，分別討論了不同臨床狀況虛擬光學切片術的角色。 我們利用倉鼠的肺部活組織檢查研究，探討虛擬光學切片術是否能夠輔助現有的工具。在此研究中，虛擬光學切片術的目的不是在取代傳統的病理切片，而是在輔助切片取樣的過程。我們發現虛擬光學切片術能夠在不破壞檢體的情況下，做檢體的虛擬光學切片檢查。檢查後的檢體，仍然可以依照一般流程送病理科檢查。我們使用6%的醋酸，增強細胞核的三倍頻，可以由虛擬光學切片術判斷檢體是否足夠，也能夠做初步的檢查。醋酸的反應是暫時且可逆的，並不影響後續的病理切片與染色。 在耳膜的研究中，我們探討了虛擬光學切片術對於不適合做傳統活組織檢查部位的應用價值。虛擬光學切片術能夠獲得耳膜的立體影像，而且能夠將顯微鏡的影像加以拼起來，合成整個耳膜的完整構造。虛擬光學切片也可以很得到耳膜中三維的纖維排列資訊，未來利用此資訊，可以輔助耳膜的手術，避免手術切斷太多的放射狀纖維，維持耳膜手術後的聽力。 在糖尿病的臨床試驗中，我們探討了非侵入式虛擬光學切片於糖尿病病患檢測的價值。在此研究中，我們成功的於不同的病患都取得良好的虛擬光學切片影像。實驗沒有造成任何的不良反應。證實了虛擬光學切片術可以是個臨床實際可用的非侵入式檢驗工具。在此研究中，針對膠原纖維的排列，我們提出了膠原纖維面積比例，膠原纖維平均方向與膠原纖維方向變異度三個新指標，雖然統計結果沒有顯著差異，但是糖尿病組的膠原纖維面積與膠原纖維排列變異度有降低的趨勢，此結果符合醫學上糖尿病會破壞膠原纖維的推論。未來仍值得持續深入研究皮膚的膠原纖維虛擬光學切片，以瞭解膠原纖維病變的嚴重程度。There are four kinds of widely used modalities in clinical medicine, X-ray, magnetic resonance imaging, ultrasound and nuclear medicine. However, the spatial resolution of these modalities is in the range of millimeters. This kind of resolution is not enough to provide information on the histological or cellular level. Recent advance of biophotonic technology enables us to obtain micrometer or even nanometer structure information in the living tissue. Under the exposure of intense laser light, there will be naturally occurring second harmonic generation and third harmonic generation signals. Using these signals, we can build a noninvasive high resolution three dimensional virtual optical biopsy system. The purpose of this thesis is to evaluate the clinical value of virtual optical biopsy using harmonic generation microscope. In the thesis, we discuss the value of virtual optical biopsy in three different kinds of clinical scenario. In the hamster lung biopsy study, we study the ability of virtual optical biopsy to aid the traditional biopsy. In this study, the role of virtual biopsy is not to replace pathology examination but to aid the clinical biopsy procedure. We found that the specimen will not be altered after virtual optical biopsy. Traditional pathology exam can be performed after virtual optical biopsy. We use 6% acetic acid to enhance the third harmonic signal from the specimen. The reaction from acetic acid is transient and reversible. The acetic acid will not influence the result of subsequent pathology examination. Using virtual optical biopsy, we can evaluate the adequacy of the specimen and obtain preliminary result. In the study of tympanic membrane, we evaluate the role of virtual optical biopsy in the tissue not suitable for traditional biopsy. We can obtain three dimensional information of tympanic membrane and can even reconstruct the whole tympanic membrane by combine multiple images. The three dimensional collagen fiber structures can be obtained from virtual optical biopsy. In the future, the surgeon may use virtual optical biopsy to guide tympanic surgery by not cutting too much of supporting radial fibers in the tympanic membrane. In this way, the complication of hearing impairment from tympanic membrane surgery will be decreased. In the clinical trial of diabetes, we study the role of noninvasive virtual optical biopsy in patient with diabetes. We successfully obtain high quality images in varies patients without any complications. This study proves that virtual optical biopsy is a clinically safe and useful modality. To quantify the collagen fiber orientation in skin, we propose three new indexes, collagen fiber area ratio, collagen fiber mean orientation and collagen fiber orientation variability. Although there is no statistically significant difference of these indices between the diabetes group and control group, there is a trend of lower collagen fiber ratio, and lower collagen fiber orientation variability in the diabetes group. This finding is compatible with the fact that there may be more collagen fiber degradation in the diabetes patients. Further study of collagen fiber should be performed. Virtual optical biopsy may provide clinically useful information on collagen fiber in skin in the future.4219948 bytesapplication/pdfen-US虛擬光學切片倍頻顯微術活組織檢查耳膜糖尿病膠原纖維Virtual optical biopsyHarmonic generation microscopyBiopsyTympanic membraneDiabetesCollagen fiber[SDGs]SDG3thesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/253883/1/ntu-101-D92921025-1.pdf