隨著顯微影像技術的突破，我們可以取得腦內神經元之間在不同的尺度下連結圖譜。神經科學家想達成的目標是了解在神經細胞中，訊號傳遞、儲存及提取的連續處理過程，第一步則是腦神經網路體建構，一個三維的腦神經網路圖譜，詳細記錄生物體的腦神經網路在結構及功能上如何連結。我們實驗室預計結合同步輻射光源及高基氏染色方法完成的全腦圖譜。我們利用了高基氏染色特性，在X-射線造影技術成熟之前，藉由減少實際染上的神經數目，達到單一神經的解析度。高基氏-卡氏染色法雖然省時，染上的神經形態卻不完整且有非預期的沉澱產生。因此為了改善高基氏-卡氏染色法品質，我們根據實驗所得的固定溫度、浸泡長度、固定緩衝液、添加劑建立一套新的染色流程，達到改善染色品質的目的。為了加速樣品製備的優化，我們採用在光學顯微鏡下利用組織澄清技術，達到初步的篩選，避免傳統包埋在塑膠的繁複過程。由同步輻射光源搭配金屬染色方法的技術進行全腦重建圖譜的技術漸趨成熟，本研究提供了高基氏染色方法優化的方向及組織澄清光學成像的快速篩選。

With the breakthrough in microscopy, we can obtain the neuron connection in the brain at either macroscale or microscale. The goal of neuroscientists is to understand how the propagation, storage, and acquisition of information are accomplished in the billions of nerve cells. First, we need to establish the wiring diagram of the brain, a three-dimensional brain mapping, the neuronal connection based on the structure and the function of nerve cells. In our lab, we will provide a synchrotron X-ray based whole-head mapping with Golgi stained specimen. The Golgi-Cox method provides a fast staining process but incomplete stained neurons as well. For the improvement of the staining quality, we establish a new protocol based on optimal experimental temperature, incubation time, fixation condition and additive. To speed the optimization of Golgi method, we decided to primarily image samples through optical microscopy with clearing agent, thioglycerol. Synchrotron X-ray based brain mapping with metallic stain technique has drawn the attention of neuroscientists. The present study introduced the potential orientations of Golgi method optimization and fast approach to screen out by clearing tissue.

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