Miller Sean J, Rothstein Jeffrey D
Department of Neurology, Johns Hopkins University School of Medicine, Johns Hopkins Univ., 855 N Wolfe Street, Room 250.18, Baltimore, MD 21205 USA.
The Brain Science Institute at Johns Hopkins, Johns Hopkins Univ. School of Medicine, Johns Hopkins Univ., 855 N Wolfe Street, Baltimore, MD 21205 USA.
Biol Proced Online. 2017 Jul 5;19:7. doi: 10.1186/s12575-017-0057-2. eCollection 2017.
Pathological analyses and methodology has recently undergone a dramatic revolution. With the creation of tissue clearing methods such as CLARITY and CUBIC, groups can now achieve complete transparency in tissue samples in nano-porous hydrogels. Cleared tissue is then imagined in a semi-aqueous medium that matches the refractive index of the objective being used. However, one major challenge is the ability to control tissue movement during imaging and to relocate precise locations post sequential clearing and re-staining.
Using 3D printers, we designed tissue molds that fit precisely around the specimen being imaged. First, images are taken of the specimen, followed by importing and design of a structural mold, then printed with affordable plastics by a 3D printer.
With our novel design, we have innovated tissue molds called innovative molds (iMolds) that can be generated in any laboratory and are customized for any organ, tissue, or bone matter being imaged. Furthermore, the inexpensive and reusable tissue molds are made compatible for any microscope such as single and multi-photon confocal with varying stage dimensions. Excitingly, iMolds can also be generated to hold multiple organs in one mold, making reconstruction and imaging much easier.
Taken together, with iMolds it is now possible to image cleared tissue in clearing medium while limiting movement and being able to relocate precise anatomical and cellular locations on sequential imaging events in any basic laboratory. This system provides great potential for screening widespread effects of therapeutics and disease across entire organ systems.
病理分析及方法学最近经历了一场巨大的变革。随着诸如CLARITY和CUBIC等组织透明化方法的出现，研究团队如今能够使纳米多孔水凝胶中的组织样本实现完全透明。随后，在与所用物镜折射率相匹配的半水介质中对透明化后的组织进行成像。然而，一个主要挑战是在成像过程中控制组织移动，以及在连续透明化和重新染色后重新定位精确位置的能力。
我们使用3D打印机设计了能够精确适配成像样本的组织模具。首先，对样本进行成像，接着导入并设计结构模具，然后用价格实惠的塑料通过3D打印机打印出来。
通过我们的新颖设计，我们创新出了名为创新模具（iMolds）的组织模具，其可以在任何实验室制作，并且能针对任何正在成像的器官、组织或骨质进行定制。此外，这种价格低廉且可重复使用的组织模具适用于任何显微镜，如具有不同载物台尺寸的单光子和多光子共聚焦显微镜。令人兴奋的是，还可以制作iMolds在一个模具中容纳多个器官，从而使重建和成像变得更加容易。
综上所述，借助iMolds，现在有可能在透明化介质中对透明化后的组织进行成像，同时限制其移动，并能够在任何基础实验室的连续成像过程中重新定位精确的解剖学和细胞位置。该系统为筛查治疗方法和疾病在整个器官系统中的广泛影响提供了巨大潜力。
