Multi-immersion Oblique Plane Microscope (miOPM): A reconfigurable platform for high-resolution Light-Sheet Fluorescence Microscopy.

Bingying Chen; Alfred Millett-Sikking; Seweryn Gałecki; Stephan Daetwyler; Jenny Jiou; Jim Monistrol; Qionghua Shen; Felix Zhou; Hsin-Yu Lin; Edward M. Jenkins; Miriam C. Stein; Madeleine Marlar-Pavey; Gabriel Sturm; Alexander D. Q. Li; Qiaosi Tang; Brian Y. Feng; Ulises Diaz; Yannan Chen; Ary Shalizi; Astrid Gillich; Jonathan R. Friedman; Raju Tomer; Bo-Jui Chang; Wallace F. Marshall; Sarah H. Shahmoradian; Kevin M. Dean; Reto Paul Fiolka

doi:10.1101/2025.10.04.680473

BioengineeringbioRxivOct 4, 2025

Preprint

Multi-immersion Oblique Plane Microscope (miOPM): A reconfigurable platform for high-resolution Light-Sheet Fluorescence Microscopy.

Bingying Chen iD, Alfred Millett-Sikking, Seweryn Gałecki iD, Stephan Daetwyler iD, Jenny Jiou iD, Jim Monistrol iD, Qionghua Shen iD, Felix Zhou iD, Hsin-Yu Lin, Edward M. Jenkins iD, Miriam C. Stein, Madeleine Marlar-Pavey iD, Gabriel Sturm iD, Alexander D. Q. Li iD, Qiaosi Tang, Brian Y. Feng iD, Ulises Diaz, Yannan Chen iD, Ary Shalizi, Astrid Gillich iD, Jonathan R. Friedman iD, Raju Tomer iD, Bo-Jui Chang iD, Wallace F. Marshall iD, Sarah H. Shahmoradian iD, , Reto Paul Fiolka iD

Abstract

Light-Sheet Fluorescence Microscopy (LSFM) enables gentle, rapid, and efficient volumetric imaging of biological specimens. Despite its potential, LSFM has fragmented into multiple variants, each narrowly optimized for specific samples or imaging regimes, limiting its broad applicability. To overcome this barrier, we introduce the multi-immersion oblique plane microscope (miOPM), an adaptable LSFM platform for high-resolution imaging ranging from subcellular dynamics to whole organisms and cleared tissues. miOPM uniquely supports seamless interchangeability of oil, water, and air objectives, and maintains diffraction limited performance across a refractive index range of 1.33 - 1.51. It is compatible with standard sample mounting, current clearing protocols and high-throughput 3D imaging. We leverage miOPM to image diverse biological specimens at multiple spatial scales. By greatly expanding the adaptability and ease of use of LSFM, miOPM is poised to democratize advanced three-dimensional imaging.

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