A versatile and customizable low-cost 3D-printed open standard for microscopic imaging

Abstract

Modern microscopes used for biological imaging often present themselves as black boxes whose precise operating principle remains unknown, and whose optical resolution and price seem to be in inverse proportion to each other. With UC2 (You. See. Too.) we present a low-cost, 3D-printed, open-source, modular microscopy toolbox and demonstrate its versatility by realizing a complete microscope development cycle from concept to experimental phase. The self-contained incubator-enclosed brightfield microscope monitors monocyte to macrophage cell differentiation for seven days at cellular resolution level (e.g. 2 μm). Furthermore, by including very few additional components, the geometry is transferred into a 400 Euro light sheet fluorescence microscope for volumetric observations of a transgenic Zebrafish expressing green fluorescent protein (GFP). With this, we aim to establish an open standard in optics to facilitate interfacing with various complementary platforms. By making the content and comprehensive documentation publicly available, the systems presented here lend themselves to easy and straightforward replications, modifications, and extensions.

UCsim2: 2D Structured Illumination Microscopy using UC2

“…This simplifies development and building of optical systems, since modules with optical functionality can then just be stacked. The original UC2 system, which is described in detail in [13], relies entirely on 3D printed cubes that host a variety of different optical components with the help of customized inserts.…”

“…The printing and assembly time of the modules becomes an issue especially during upscaling towards multiple parallel setups. Furthermore, the designs presented in our previous publication [13] suffer from slight differences in scale between different 3D-printers with the stability and reproducibility depending also on the choice of the printing material.…”

“…This mechanism automatically centres the part and avoids mechanical ambiguities. During the designing process, we ensured that the new version is downward-compatible with previously presented 3D-printable cubes [13]. By using M5 set screws in the positions of the casted pins, this novel positive locking mechanism can also be used with fully 3D-printed modules, even though this design is not optimized for printing.…”

“…Likewise, a MEMS-based setup [12] creating an incoherently temporally varying excitation pattern could demonstrate a simple realisation of an ISM, which reduces complexity and costs by relying on commercially available laser video projectors. With the modularization and the resulting simplification of optical setups, we have recently been able to demonstrate with the open-source optical toolbox UC2 [13] that sophisticated setups can also be accomplished outside an optical laboratory by educated users from other fields.…”

The Field Guide to 3D Printing in Microscopy

“…2). These characteristics enable systems such as miCube (13), the UC2 (14), and the the Warwick Open Source Microscope (WOSM) (https://wosmic.org), where further customisation is possible due to their modular design. The microscope parts that can be built vary with the technology and materials available.…”