A 3D printed modular microscope for in-incubator time-lapse fluorescence imaging
Year: 2026
Type: article
Abstract: We aim to develop a low-cost fluorescence microscope capable of obtaining time-lapse images of cell biology processes in vivo within a cell incubator setting. The commercial systems for fluorescence microscopy are expensive and require extensive setup for use in incubators. In recent years, 3D printing technology has enabled affordable alternatives to traditional frame structures and rigs, based on which multiple open-source frameworks have emerged for building low-cost, laboratory-grade microscopes. We used the open-source UC2 framework to develop a modular microscope tailored to fluorescent imaging within an incubator. We adopted standardized modules for the motor-controlled objective z-stepper, the Raspi camera, and a mirror, and developed custom-built modules for the laser, neutral density filter, beam expander, dichroic beamsplitter, tube lens, and sample holder. The initial version was built for benchtop imaging in a minimalist approach containing only a sample holder, objective, and Raspi camera module. This allowed us to optimize the software for controlling the motor to focus the objective. In subsequent iterations, we developed customized drop-in modules for fluorescent imaging. The current version features infinity optics, allowing placement of additional optical components between the camera and the objective. These modules can be combined in multiple ways based on the physical constraints of the incubator and the desired light path. The assembled microscope is controlled by a Raspberry Pi device. We developed Python code to control the objective z-stepper and the laser, as well as to coordinate between the two modules for automated time-lapse image capture. We are testing the system to image the cellular response to wounding in an ex vivo chick post-cataract surgery explant system, an established experimental model for studying the lens response to cataract surgery-induced wounding. We are visualizing cell boundaries using fluorescently conjugated Wheat Germ Agglutin, a lectin that demarcates cell membranes by binding GlcNAc and sialic acid residues. We are collecting images of the lens cellular response to wounding over 3 days, a typical period for wound closure. In summary, we demonstrate a modular, reconfigurable, 3D-printed microscope custom-built for in-incubator time-lapse fluorescence imaging for cell biology applications.
Institution: Thomas Jefferson University
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W7219732177
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