超高解析光學同調斷層掃描於單顆皮膚細胞之造影與分析

Imaging and Analysis of Single Skin Cell by Ultrahigh-Resolution Optical Coherence Tomography

謝宗勳(Tsung-Hsun Hsieh)

指導教授 ： 黃升龍

國立臺灣大學/電機資訊學院/光電工程學研究所/碩士(2012年)

https://doi.org/10.6342/NTU.2012.00072

摘要

超高解析光學同調斷層掃描(ultrahigh-resolution optical coherence tomography; UR-OCT)首次應用於體外單顆皮膚細胞的三維造影與分析研究上。此非侵入式、毋須標定、以及擁有極深成像與細胞等級之空間解析能力的光學偵測技術，可從三維的方式來分析單顆細胞之散射特性。基於UR-OCT影像以及細胞形態學上的觀察，我們可以很容易地辨別活的和細胞凋亡之基底癌細胞。除此之外，我們首次提出一種全新且快速之自動提取個別單顆皮膚細胞之特徵參數的方法，包含訊號平均、細胞體積、細胞密度及平均動態範圍。並藉此分析方法進行了定量比較和參數分析，其中三個特徵參數p-value小於0.05。實驗結果顯示出UR-OCT於細胞級別上具有檢測細胞死亡與否之能力。
更進一步，我們藉由圖像分割中屬於型態學影像處理之膨脹與侵蝕技術，進行一系列的影像處理，並計算出由UR-OCT拍攝的HaCaT之細胞大小和細胞核體積，進而推算出細胞核質比為0.18，非常接近螢光顯微鏡之結果。
最後，我們採用HaCaT與Hs68細胞株作為表皮中的角質細胞與真皮中的纖維母細胞代表，同樣使用自行定義之特徵參數，藉由圖像分割中的k-means聚類法，從統計學上對不同類型的皮膚細胞進行區分。我們展現了總共12顆的HaCaT與Hs68細胞可以很準確且快速地被辨別，暗示了UR-OCT於未來皮膚科臨床應用與組織工程上扮演一個重要角色。

關鍵字

光學同調斷層掃描 ； 單顆細胞 ； 基底癌細胞 ； 細胞凋亡

並列摘要

Ultrahigh-resolution optical coherence tomography (UR-OCT) has been used to study single skin cells in vitro for the first time. This non-invasive and label-free optical detection technique with deep imaging depth can be used to analyze scattering properties of single cells in three-dimensional with cellular spatial resolution. Based on morphological observation of the UR-OCT images, live and apoptotic basal cell carcinoma (BCC) can be easily identified. In addition, we developed a novel method to automatically extract the characteristic parameters which defined as signal average, cell volume, cellular density and average dynamic range of individual cells. Quantitative comparison showed that three parameters were found with p-value smaller than 0.05. The experimental results show that the ability of UR-OCT to detect cell death at cellular level.
To find the nuclear-cytoplasmic ratio (N:C ratio) of single cells, we did a series of image segmentation processes by means of morphological dilation and erosion. In this way, calculated N:C ratio of single HaCaT cell was 0.18, which was very close to the observation of confocal fluorescence microscopy.
Finally, the HaCaT and Hs68 cell lines were used to model epidermis and dermis respectively. We applied clustering analysis on these cells by k-means clustering method based on the characteristic parameters. We demonstrate that a total of 12 HaCaT and Hs68 cells can be distinguished very quickly and accurately, indicating that the UR-OCT will play an important role in clinical dermatology and tissue engineering in the future.

並列關鍵字

Optical coherence tomography ； Single cell ； Basal cell carcinoma ； Apoptosis

參考文獻

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