利用組織切片技術研究腦組織三維結構

Abstract

前腦發育畸形症屬稀罕人腦疾病，胎兒發育期間會發展出不正常的腦室，可依據不同的腦室變形程度加以分類。得此病最嚴重的情況胎兒只會有一個腦室，無法順利發育雙眼與鼻子。通常胎兒發育期間最先會由醫學超音波檢查診斷出該疾病，在最嚴重的情況下出生後無法存活。根據病理學，研究異常人體器官取下之組織樣本時須將組織切片並且由光學顯微鏡在小於釐米尺度的範圍觀察切片的細節。在本研究中，我們利用連續石蠟切片染色後由高解析度白光顯微鏡分析組織切片，並試著比較正常與前腦發育畸形之胎兒腦組織在二維及三維結構上的差異。另外我們也希望透過新型振動式切片技術與雙光子顯微技術的結合發展出更好的腦組織三維完整取像方法。使用任何方式進行組織切片時都會有組織損失的問題，進而增加三維影像接合之變數，本文亦將探討此問題對三維結構重建之影響。

Holoprosencephaly, a kind of brain disease, is usually identified by ultrasonography on fetuses during gestation and confirmed by Magnetic Resonance Imaging (MRI). In the most severe cases the fetus carries craniofacial malformations Cyclopia and Proboscis above the eye with the nose absent. In this research we use the standard histological method to study the microscopic anatomy of brain tissue from human postmortem fetal brain with the Holoprosencephaly disease. Consecutive paraffin slices stained with Hematoxylin and Eosin were imaged by light microscopy and the images were then reconstructed into 3D stacks for more information on the anatomy of the disease. We also study the possibility of using agarose for embedding brain tissue along with the oscillating blade cutting technique to obtain better 3D image from slices with two-photon microscopy.