Miller James Christian, Koirala Prashanna, Argote de la Torre Maria Fernanda, Farsi Marjan, Lieberth Jaret, Shrestha Rabina, Bloomekatz Joshua
Department of Biology, University of Mississippi, University MS 38677.
bioRxiv. 2025 Jun 3:2025.06.01.657246. doi: 10.1101/2025.06.01.657246.
Embryo mounting is one of the technical challenges researchers encounter when undertaking an imaging project. Embryos need to be oriented in a reproducible manner such that the tissue of interest is accessible to a microscope objective for the entire imaging period. To overcome this challenge researchers can embed embryos in viscous media or create specialized dishes and casts to hold embryos in a desired orientation during imaging. Here we describe a method for using a cheap stereolithographic (SLA) 3D-printer to manufacture re-usable molds that create agarose wells in which embryos can be mounted for imaging. These agarose wells provide a reliable means for orienting multiple embryos for imaging. This method includes a design framework that can be easily customized for a variety of tissues, organisms and imaging challenges. Using this method we have created molds for imaging cardiac development in zebrafish for both upright and inverted microscopes. By utilizing materials and equipment that are accessible this method allows researchers to easily create molds specific to their mounting needs.
胚胎固定是研究人员在进行成像项目时遇到的技术挑战之一。胚胎需要以可重复的方式进行定向，以便在整个成像期间，感兴趣的组织能够被显微镜物镜观察到。为了克服这一挑战，研究人员可以将胚胎嵌入粘性介质中，或者制作专门的培养皿和铸型，以便在成像过程中将胚胎保持在所需的方向。在这里，我们描述了一种使用廉价的立体光刻（SLA）3D打印机制造可重复使用模具的方法，该模具可以制作琼脂糖孔，胚胎可以在其中固定以进行成像。这些琼脂糖孔为多个胚胎的成像定向提供了可靠的方法。该方法包括一个设计框架，可以很容易地针对各种组织、生物体和成像挑战进行定制。使用这种方法，我们已经为斑马鱼心脏发育成像制作了模具，适用于正立和倒置显微镜。通过使用容易获得的材料和设备，这种方法使研究人员能够轻松制作符合其固定需求的模具。
