Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, IL, USA.
Committee on Development, Regeneration, and Stem Cell Biology, The University of Chicago, Chicago, IL, USA.
Methods Mol Biol. 2023;2626:277-289. doi: 10.1007/978-1-0716-2970-3_14.
The Drosophila egg chamber is a powerful system to study epithelial cell collective migration and polarized basement membrane secretion. A strength of this system is the ability to capture these dynamic processes in ex vivo organ culture using high-resolution live imaging. Ex vivo culture also allows acute pharmacological or labeling treatments, extending the versatility of the system. However, many current ex vivo egg chamber culture setups do not permit easy medium exchange, preventing researchers from following individual egg chambers through multiple treatments. Here we present a method to immobilize egg chambers in an easy-to-construct flow chamber that permits imaging of the same egg chamber through repeated solution exchanges. This will allow researchers to take greater advantage of the wide variety of available pharmacological perturbations and other treatments like dyes to study dynamic processes in the egg chamber.
果蝇卵子发生腔是研究上皮细胞集体迁移和极化基底膜分泌的强大系统。该系统的一个优势是能够使用高分辨率活细胞成像在体外器官培养中捕获这些动态过程。体外培养还允许进行急性药理学或标记处理，从而扩展了该系统的多功能性。然而，许多当前的体外卵子发生腔培养设置不允许进行简单的培养基交换，这使得研究人员无法通过多次处理来跟踪单个卵子发生腔。在这里，我们提出了一种将卵子发生腔固定在易于构建的流动腔中的方法，该方法允许通过重复的溶液交换来对同一卵子发生腔进行成像。这将使研究人员能够更好地利用广泛的药理学干扰和其他处理方法（如染料）来研究卵子发生腔中的动态过程。
