A 3D Built-in Thermostatized Chamber for Fluorescence Microscopy
AbstractRapid prototyping using 3D printing has increased its popularity by allowing generation of small devices in a single piece or as built-in assemblies within hours. In optical microscopy, this technology has improved the design of cell culture dishes and chambers for orienting and immobilizing animal embryos. Generally, these devices are custom designed to support multiple specimens for screening employing objectives of low magnification. Varying size and geometry of developing embryos pose a significant challenge for imaging. In addition, temperature control of the incubation media is not trivial. However, few protocols make these options available. This work deals with the construction and performance of an inexpensive 3D-printed water-recirculating chamber adapted to the stage of an inverted microscope for imaging zebrafish embryos at controlled temperatures. We explored the possibility of heating and cooling the recirculating water using a Peltier cell. Although the system reached the temperatures set in the incubation media, its control over time was limited. The 3D built-in device is though versatile regarding embryo mounting and manipulation within the microscope stage, and allowed the use of high resolution objectives with minimal risk of water leakage into the optics.