Researchers at Bielefeld University in Germany have made a groundbreaking development in the field of microscopy with the creation of a fluorescence microscope that utilizes structured illumination. This innovative microscope has the ability to conduct fast super-resolution imaging across a wide field of view, allowing for the simultaneous imaging of multiple living cells. The primary goal of this microscope is to enable researchers to study and understand the effects of various drugs and drug combinations on the human body, particularly in the context of polypharmacy.

Polypharmacy refers to the administration of multiple drugs to chronically ill or elderly patients. This common practice can have dangerous consequences due to potential drug interactions. The microscope developed as part of the EIC Pathfinder OpenProject DeLIVERy seeks to shed light on this issue by providing a platform for investigating polypharmacy in individual patients. By enabling the imaging of isolated living cells with super-resolution capabilities, researchers can observe the dynamics of cell membrane feature and organelles when exposed to different drug combinations.

Published in the journal Optics Express, the researchers describe the features and capabilities of their fluorescence microscope. It utilizes optical fiber delivery of excitation light to achieve high image quality over a large field of view, all while maintaining a high resolution. The microscope is capable of imaging liver cells with a field of view of up to 150 x 150 μm2 and imaging rates of up to 44 Hz, while still achieving a spatiotemporal resolution of under 100 nm. These impressive specifications make it a highly effective tool for studying the effects of drugs on cells.

The foundation of this novel microscope lies in super-resolved structured illumination microscopy (SR-SIM), which involves the use of a structured pattern of light to excite fluorescence in a sample beyond the diffraction limit of light. SR-SIM is particularly well-suited for live cell imaging as it utilizes low-power excitation that does not harm the sample, all while producing highly detailed images. With the ability to achieve high resolution across a wide field of view, the new microscope is capable of reconstructing super-resolved images using a set of raw images.

To achieve high resolution across a wide field of view, the microscope’s image reconstruction relies on a set of optical fibers that illuminate the sample with a sinusoidal striped pattern. By shifting and rotating this pattern, additional information is obtained and used to reconstruct super-resolved images. The use of a newly designed fiber switch and a hexagonal holder allows for precise adjustment of all beams, resulting in a large field of view that is compatible with live-cell imaging. The setup is versatile enough to be used for total internal reflection fluorescence excitation (TIRF)-SIM, which restricts fluorescence excitation and detection to a thin region of the sample.

Given that the liver plays a critical role in drug metabolism, the researchers tested the microscope using samples of fixed multicolor-stained rat liver cells. The reconstructed images produced by the microscope showcased the visualization of tiny membrane structures that are smaller than the diffraction limit of light. The development of this compact system has revolutionized the field by combining a large field of view, fast pattern switching speed, and multicolor, power-efficient excitation. Additionally, the setup allows for either 2D-SIM or TIRF-SIM imaging, further enhancing its versatility and practicality.

Moving forward, the researchers plan to leverage the capabilities of this microscope by conducting live cell studies of liver cells. This will allow them to observe the dynamics of cells when treated with different drugs. Additionally, efforts are underway to improve the image reconstruction process, with the ultimate goal of achieving real-time reconstruction of acquired raw data. The development of this groundbreaking fluorescence microscope has opened up new avenues of research, paving the way for advancements in personalized healthcare and clinical applications that require high-resolution imaging.

The development of a fluorescence microscope utilizing structured illumination has revolutionized the field of microscopy. This innovative instrument provides researchers with the ability to conduct fast super-resolution imaging over a wide field of view, enabling the simultaneous imaging of multiple living cells. By studying the effects of various drugs and combinations of drugs on cells, researchers hope to gain valuable insights into the dangers of polypharmacy and improve personalized healthcare. As technology advances and image reconstruction processes improve, the potential applications of this microscope are boundless, holding great promise for the future of scientific research and medical advancements.

Physics

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