Department of Microbiology and Immunology, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON N6A 5C1, Canada.
Robarts Research Institute, London, Ontario, Canada.
Biochem Cell Biol. 2020 Oct;98(5):612-623. doi: 10.1139/bcb-2020-0064. Epub 2020 Apr 27.
Using multiple imaging modalities while performing independent experiments in parallel can greatly enhance the throughput of microscopy-based research, but requires the provision of appropriate experimental conditions in a format that meets the optical requirements of the microscope. Although customized imaging chambers can meet these challenges, the difficulty of manufacturing custom chambers and the relatively high cost and design inflexibility of commercial chambers has limited the adoption of this approach. Herein, we demonstrate the use of 3D printing to produce inexpensive, customized, live-cell imaging chambers that are compatible with a range of imaging modalities, including super-resolution microscopy. In this approach, biocompatible plastics are used to print imaging chambers designed to meet the specific needs of an experiment, followed by adhesion of the printed chamber to a glass coverslip, producing a chamber that is impermeant to liquids and that supports the growth and imaging of cells over multiple days. This approach can also be used to produce moulds for casting microfluidic devices made of polydimethylsiloxane. The utility of these chambers is demonstrated using designs for multiplex microscopy, imaging under shear, chemotaxis, and general cellular imaging. Together, this approach represents an inexpensive yet highly customizable approach for producing imaging chambers that are compatible with modern microscopy techniques.
在并行进行独立实验时使用多种成像模式可以大大提高基于显微镜的研究的通量，但需要以满足显微镜光学要求的格式提供适当的实验条件。虽然定制成像室可以满足这些挑战，但制造定制室的难度以及商业室的相对较高的成本和设计灵活性限制了这种方法的采用。在这里，我们展示了使用 3D 打印来生产廉价的、定制的、用于活细胞成像的腔室，这些腔室与多种成像模式兼容，包括超分辨率显微镜。在这种方法中，使用生物相容性塑料来打印成像腔室，这些腔室设计旨在满足实验的具体需求，然后将打印腔室粘附到玻璃盖玻片上，产生一种对液体不可渗透的腔室，并支持细胞在数天内的生长和成像。这种方法也可用于制作聚二甲基硅氧烷微流控器件的铸造模具。使用用于多路显微镜、剪切成像、趋化性和一般细胞成像的设计来证明这些腔室的实用性。总之，这种方法代表了一种廉价但高度可定制的生产与现代显微镜技术兼容的成像腔室的方法。
