Lee Boram, Lee Eunsoo, Kim June Hoan, Kim Hyung-Jin, Kang Yong Guk, Kim Hyun Jung, Shim Jin-Kyoung, Kang Seok-Gu, Kim Beop-Min, Kim Karam, Kim Youseok, Cho Keunchang, Sun Woong
Department of Anatomy, Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Seoul 02841, Republic of Korea.
Fluorescence Core Imaging Center, Ewha Womans University, Seoul 03760, Republic of Korea.
iScience. 2021 Mar 5;24(4):102267. doi: 10.1016/j.isci.2021.102267. eCollection 2021 Apr 23.
Optical clearing has emerged as a powerful tool for volume imaging. Although volume imaging with immunostaining have been successful in many protocols, yet obtaining homogeneously stained thick samples remains challenging. Here, we propose a method for label-free imaging of brain slices by enhancing the regional heterogeneity of the optical properties using the tissue clearing principle. We used FxClear, a method for delipidation of brain tissue, to retain a larger proportion of lipids at the white matter (WM). Furthermore, the embedding media affected the contrasts for the lipid-rich or extracellular matrix-rich areas, depending on their chemical properties. Thus, we tailored clearing conditions for the enhancement of the refractive indices (RIs) differences between gray and WM, or several pathological features. RI differences can be imaged using conventional light microscopy or optical coherence tomography. We propose that our protocol is simple, reliable, and flexible for label-free imaging, easily implementable to routine histology laboratory.
光学透明化已成为一种用于容积成像的强大工具。尽管在许多方案中，免疫染色的容积成像已取得成功，但获得均匀染色的厚样本仍然具有挑战性。在此，我们提出一种通过利用组织透明化原理增强光学特性的区域异质性来对脑切片进行无标记成像的方法。我们使用FxClear（一种用于脑组织脱脂的方法）在白质（WM）中保留较大比例的脂质。此外，包埋介质根据其化学性质影响富含脂质或富含细胞外基质区域的对比度。因此，我们调整了透明化条件以增强灰质和白质之间的折射率（RI）差异或几种病理特征。RI差异可以使用传统光学显微镜或光学相干断层扫描进行成像。我们认为我们的方案对于无标记成像而言简单、可靠且灵活，易于在常规组织学实验室中实施。
