利用三維折射率顯微術定量式分析紅血球細胞

貧血為一種常見的人體疾病，可以看成是一種單一臨床上的疾病也可能是某種慢性疾病的症狀之一，這是由於血液當中的紅血球攜帶氧氣的能力不足，無法正常地將氧氣輸送到人體各個器官，所產生的一種症狀。透過抽血進行血液檢驗，藉由全血自動分析儀計算出血液中的紅血球、白血球、血小板等相關的數值是否介於人體的正常參考區間內。檢驗是否有貧血症狀目前可以從紅血球體積以及紅血球內的血紅素含量作為初步的判斷依據，目前臨床上使用的全自動血液分析儀在紅血球細胞的測量方式，大多採用直流電阻抗技術法去計算細胞的體積以及數量，此方法的優點可以在短時間內測量大量細胞平均結果，速度快、信息量大，但無法比較單一紅血球細胞之間的差異性以及提供型態上的判讀。因此，全自動血液分析儀在初步檢驗上是一項很好的工具，但仍需仰賴後續的人工處理分析進一步情況。基於實驗室所開發的三維折射率顯微鏡(術)，透過干涉全像術以及電腦斷層影像的概念，重建出細胞的三維折射率分布，即為細胞的三維結構。將此方法應用在紅血球細胞上，可以重建出紅血球的三維折射率分布，並量化單一細胞的體積以及血紅素含量，更可以同時觀察細胞間的型態差異。透過此系統，將可以在無需染色標定的情況下，得到紅血球多項的內生性資訊，如體積、血紅素濃度、折射率值分布以及三維結構等等。藉由三維折射率顯微鏡的測量，我們已經初步定量出正常以及貧血疾病的紅血球在各項數值上的結果，並與全自動血液分析儀測得的臨床數值做比較。目前統計結果在體積以及濃度表現上雖與全自動血液分析儀有所差異，但仍呈現出正相關的結果，而從細胞折射率值的表現上，也能夠看出正常及貧血兩者之間存在明顯的不同，並能清楚的表現出紅血球細胞的三維型態。現階段由於細胞統計的樣本數量較少，在病患上也只有輕微的地中海型貧血的資料，因此在未來希望能量化出更多的紅血球細胞，及提升系統的準確性，獲得更為可信的統計結果。

關鍵字

紅血球；血紅素；貧血；全像干涉術；三維折射率影像

並列摘要

Anemia is a common disease caused by low amount of red blood cells (RBCs)/hemoglobin in the blood, or a lowered ability of the blood to carry oxygen. The symptoms include fatigue, dyspnea, confusion or syncope. Volume and hemoglobin contents of the RBCs are the indices to determine if the patient has the anemia symptom in the initial blood test. The current method in the clinical hospital is the fully-automated whole blood analyzer for calculating the indices of RBCs. Using this method, the RBCs analysis can be finished in a short time and acquire high throughput data. The fully-automated whole blood analyzer is good for RBCs clinical analysis in the initial blood test although it also needs more manual process by clinical laboratory technologist when the machine detects the abnormal state on the blood. Furthermore, the automated machine cannot provide morphology of the RBCs. The goal of the thesis is to establish a method that can reconstruct 3D morphology of RBCs and quantify the volume and hemoglobin contents by a novel optical microscopy technique: three-dimensional (3D) refractive-index (RI) microscopy which is based on digital holography and optical diffraction tomography. Using this technique, we can reconstruct the 3D-RI mapping of the BRCs and evaluate the deviation between individual RBCs without staining or other extra process. 3D-RI microscopy have been used for the measurement of normal and thalassemia RBCs and the statistical result of volume and Hb concentration are compared with the clinical blood tests. The 3D-RI microscopy was practically performed on mapping 3D-RI distribution of RBCs and we found that the RI and morphology difference between normal and the thalassemia patients are clear to be observed in our results. In the future, we will measure more RBCs from normal and anemia patients with different symptoms to establish a more reliable statistics.

並列關鍵字

Red ； blood ； cells ； imaging ； hemoglobin ； anemia ； holography ； three-dimensional refractive index image

參考文獻

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利用三維折射率顯微術定量式分析紅血球細胞

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