- 學位論文
利用雙光子顯微術以及動態模型觀測肝代謝功能
Utilizing Multiphoton Microscopy and Kinetic Model to Monitor In Vivo Metabolic Activity of the Liver
摘要
肝乃是人體最重要的器官之一，而慢性乾病變長年在亞洲位居十大死因之榜上，因此對於肝臟的研究刻不容緩。然而肝是沈默的器官，缺乏神經來表達它的狀態，只能透過外在的觀察來判斷肝的狀況。現行的檢查技術，例如：GPT、GOT、ICG、CT、MRI…等等，固然已經幫人類解決很多問題了，但其共同點為：缺乏一個活體顯微以及動態及時的觀測。 利用雙光子螢光顯微鏡以及肝視窗，我們可以看見動態螢光分子（6-CFDA），在肝臟中的即時動態。而解析度可以進一步的到達細胞（微米）等級。同時我們發現，肝臟代謝6-CFDA的過程，可以藉由兩個第一階速率方程（first-order rate equation）來描述。利用微分方程的解，來模擬螢光變化，我們以此量化肝臟攝取（k1）以及排出（k2）6-CFDA的能力。並藉由比較此二參數，來定義乾細胞的正常與否，或者整體肝臟的狀態。同時探討參數的分布狀況，可以看出肝小葉中細胞能力的分布，並討論其原因。 最終的目的在於：希望可以藉由直接觀察描述顯微活體影像，來連結其他方法所觀測到的巨觀病理狀況。
並列摘要
Liver is one of the most important organs in our bodies. Meanwhile, liver disease is one of the leading causes of death in Asia, especially Taiwan. Study of liver is important and urgent. Unfortunately, liver is one of the organs lacking neuro system to report its situation. Diagnoses of liver, such as GPT, GOT, ICG, CT, MRI…etc. already solved lots of problem for human being. One of their common drawback is they don’t have real-time and in vivo microscopic analysis. Utilizing two-photon fluorescent microscopy (TPFM) and liver chamber. We would able to monitor real-time in vivo image of movement of fluorescent probe, 6-CFDA. At the same time, we found that the metabolism of 6-CFDA could be expressed by two first-order rate equations. These equations helped us to quantify ability of influx (k1) and efflux (k2) of hepatocytes. By comparing these for parameters, we’d like to define cells’ situations or stages of whole liver. Meanwhile, zonation difference could be revealed by discussion of distribution of both parameters. Finding the connection between macroscopic pathology and histology in previous studies and in vivo microscopic situation we monitored is our ultimate purpose.