使用層光顯微鏡量化斑馬魚房室通道動態結構在不同酒精濃度下的影響

Abstract

本實驗主要針對斑馬魚心臟房室通道做進一步的量化探討，房室（AV）瓣膜是心臟跳動節奏形成中的關鍵組成部分，並且在預防心室收縮期間從心室向心房逆流方面起著至關重要的作用。以往許多研究和臨床研究已經指出人類和脊椎動物模型的房室瓣膜故障或畸形會導致心髒病的發病原因，因此測量AV通道運動的方法可以潛在地解決心臟中的許多問題。 我們針對斑馬魚研究架設一套層光螢光顯微鏡系統，並且設計一套房室通道量化方法，將層光螢光顯微鏡系統所得到的影像，做量化以及FFT分析。最後使用顯微注射劑注射不同弄度酒精至斑馬魚血管內部，用此光學系統以及量化方法來分析斑馬魚在不同濃度酒精下房市通道跳動反應。 本實驗目的在於開發一種LSFM光學系統量化方法，此方法可以評估斑馬魚房室通道健康狀態重要指標，進而瞭解房室瓣膜病變機制，未來希望能更進一步做藥物篩檢和毒物檢測。

This research explores optical quantification of the structural dynamics of zebrafish atrioventricular (AV) canal which is a key component in the formation of heart beating rhythm and vital for prevention of backflow of blood from ventricle to atrium under ventricular contraction. Previous research and clinical studies have indicated congenital or acquired pathogenesis of canal diseases attributed to malfunction or malformation of AV valve in both human and vertebrate animal models. Therefore, the development of a method that measures valvular dynamics of zebrafish may potentially facilitate drug screening of human implanted diseases. Our approach is to setup a light-sheet fluorescence microscopy(LSFM) in combination with numerical quantitative method to quantify the structural dynamics of AV canal. Specifically the time-sequenced images obtained by LSFM is numerical quantified with a threshold method and analyzed by a built-in function of fast fourier transform in Matlab. Finally, LSFM is assessed by microinjection of different percentile content of alcohol into the blood vessel of zebrafish to elucidate quantitatively how the width of AV canal of zebrafish responds to the injection of alcohol. For the prospects of future work, the system will be optimized for its resolution, applied to examined human implanted diseases and drug screening for therapeutics of cardiac disorders.