超快雷射的發明提供多光子激發螢光顯微術在生物醫學相關領域上多樣性的應用，發展至今解析度與掃描幀率不斷地提升，共軛顯微術大幅提升了三維影像對比度；高速掃描器提供更快的影像幀率，因此許多研究欲觀測活體細胞動態便需要透過即時觀測系統幫助。本論文主要目的為建構以快速二維掃描為基礎開發高時間空間解析度之多光子顯微術，並將其應用在動態分子影像上，希望提升二維影像幀率。將以往雙軸掃描系統中，其中一軸震鏡改為共振掃描震鏡(resonant scanning galvanometer)，其振盪頻率為8 kHz，提供更快速的點掃描速度，搭配同公司出產之Pixel Clock電路板，能擷取共振掃描震鏡掃描資訊，其像素同步頻率最高達15 MHz，為應付如此快速的取樣速度，使用由NI公司出產之FlexRIO介面卡和高速資料擷取前端模組，其中包含現場可程式化閘陣列(Field-programmable gate array，FPGA)，可快速收取兩軸震鏡信號並觸發光電倍增管擷取螢光訊號，使用NI公司成熟發展的LabVIEW軟體設計整體系統程式介面，處理所有類比數位訊號、讀取數據以及呈現多光子影像。

The invention of ultrafast-laser provides various applications for multiphoton microscopy in biomedical field. Recently, the resolution and frame rate of microscopy have great improvements. Confocal microscopy increases the 3D image contrast. The high speed scanning tools provide faster frame rate. In this research, we build a high temporal and spatial resolution multiphoton microscope based on high-speed 2D scanning method and we hope to increase frame rate for imaging cell dynamics. We adopt resonant galvanometer whose resonant frequency is 8 kHz to achieve rapid scanning speed in order to replace the traditional galvanometer. The system combines with the pixel clock board with the frequency ranges from 7.5 MHz to 15 MHz that contains scanning information of resonant galvanometer. Due to the fast sampling rate, we utilize FlexRIO which contains field-programmable gate array (FPGA) chip and high-speed data acquisition module to collect signal from galvanometers and to trigger photomultiplier tubes (PMT) to receive fluorescence signal. We use LabVIEW programming language of National Instruments (NI) to design overall system to process analog and digital signals, system parameters, and display images.

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