Palmer William M, Flynn Jamie R, Martin Antony P, Reed Stephanie L, Grof Christopher P L, White Rosemary G, Furbank Robert T
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW, Australia.
School of Biomedical Sciences and Pharmacy, University of Newcastle, Callaghan, NSW, Australia.
Methods Mol Biol. 2018;1770:285-304. doi: 10.1007/978-1-4939-7786-4_17.
Plant histology and imaging traditionally involve the transformation of tissues into thin sections to minimize light scatter in opaque material, allowing optical clarity and high-resolution microscopy. Recently, new techniques in 3D tissue clearing, including PEA-CLARITY, have been developed to minimize light scatter within intact, whole samples. These techniques can achieve equivalent microscopic resolution to that of thin section imaging with the added benefit of maintaining the original 3D structure and position of biomolecules of interest. Furthermore, PEA-CLARITY is compatible with standard stains and immunohistochemistry, allowing molecular interrogation of intact, 3D tissues. This chapter outlines the current methods available for 3D histology in plants and details the materials, equipment, reagents, and procedure for the PEA-CLARITY technique.
植物组织学和成像传统上涉及将组织转化为薄片，以尽量减少不透明材料中的光散射，从而实现光学清晰度和高分辨率显微镜观察。最近，包括PEA-CLARITY在内的三维组织透明化新技术已经被开发出来，以尽量减少完整全样本内的光散射。这些技术可以实现与薄片成像相当的微观分辨率，同时还具有保持感兴趣生物分子的原始三维结构和位置的额外优势。此外，PEA-CLARITY与标准染色和免疫组织化学兼容，允许对完整的三维组织进行分子检测。本章概述了目前可用于植物三维组织学的方法，并详细介绍了PEA-CLARITY技术的材料、设备、试剂和步骤。
