BACKGROUND 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. METHODS 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. RESULTS 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. CONCLUSIONS 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.

中文翻译：

---

3D打印机生成的组织iMolds用于清除组织，使用单光子和多光子显微镜进行深层组织评估。

背景技术病理学分析和方法论最近经历了戏剧性的革命。通过创建诸如CLARITY和CUBIC之类的组织清除方法，小组现在可以在纳米多孔水凝胶中的组织样品中实现完全透明。然后在与所用物镜的折射率相匹配的半水介质中想象被清除的组织。然而，一项主要挑战是在成像过程中控制组织运动并在顺序清除和重新染色后重新定位精确位置的能力。方法我们使用3D打印机设计了可以精确地围绕要成像的标本的组织模具。首先，获取标本的图像，然后导入和设计结构模具，然后通过3D打印机用负担得起的塑料进行打印。结果凭借我们新颖的设计，我们开发了创新的组织模具，称为创新模具（iMolds），可以在任何实验室中生成，并且可以针对要成像的任何器官，组织或骨骼进行定制。此外，使便宜且可重复使用的组织模具与任何显微镜兼容，例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，还可以生成iMolds以将多个器官固定在一个模具中，从而使重建和成像更加容易。结论与iMolds一起使用，现在可以在清除介质中对清除的组织进行成像，同时限制移动，并能够在任何基础实验室中按顺序进行的成像事件重新定位精确的解剖和细胞位置。该系统具有很大的潜力，可用于筛选整个器官系统中治疗剂和疾病的广泛影响。