Neuroscience and Behavior Graduate Program, University of Massachusetts Amherst, Amherst, USA.
Department of Biology, University of Massachusetts Amherst, Amherst, USA.
Sci Rep. 2021 Aug 12;11(1):16462. doi: 10.1038/s41598-021-95692-2.
Techniques used to clear biological tissue for fluorescence microscopy are essential to connect anatomical principles at levels ranging from subcellular to the whole animal. Here we report a simple and straightforward approach to efficiently render opaque tissue samples transparent and show that this approach can be modified to rapidly label intact tissue samples with antibodies for large volume fluorescence microscopy. This strategy applies a magnetohydrodynamic (MHD) force to accelerate the removal of lipids from tissue samples at least as large as an intact adult mouse brain. We also show that MHD force can be used to accelerate antibody penetration into tissue samples. This strategy complements a growing array of tools that enable high-resolution 3-dimensional anatomical analyses in intact tissues using fluorescence microscopy. MHD-accelerated clearing is simple, fast, reliable, inexpensive, provides good thermal regulation, and is compatible with existing strategies for high-quality fluorescence microscopy of intact tissues.
用于荧光显微镜的生物组织清除技术对于将解剖学原理从亚细胞水平连接到整个动物水平至关重要。在这里,我们报告了一种简单而直接的方法,可以有效地使不透明的组织样品透明,并表明可以修改该方法以快速用抗体标记完整的组织样品,用于大容量荧光显微镜。该策略应用磁流体动力学(MHD)力来加速去除组织样品中的脂质,其大小至少与完整的成年小鼠大脑一样大。我们还表明,MHD 力可用于加速抗体渗透到组织样品中。该策略补充了一系列越来越多的工具,这些工具可使用荧光显微镜在完整组织中进行高分辨率的 3 维解剖分析。MHD 加速清除方法简单、快速、可靠、经济实惠,提供良好的热调节,并与现有的完整组织高质量荧光显微镜策略兼容。