biostudies-literatureUnknownChen BNIBIB NIH HHSNIMH NIH HHSNCI NIH HHSNIGMS NIH HHSLight-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.bioRxiv : the preprint server for biology2025.10.04.680473https://www.ebi.ac.uk/biostudies/studies/S-EPMC12632540biostudies-literatureMulti-immersion Oblique Plane Microscope (miOPM): A reconfigurable platform for high-resolution Light-Sheet Fluorescence Microscopy.PMC12632540DP2 MH119423RM1 GM145399R35 GM137894R01 EB035538R35 GM119768U54 CA268072R35 GM133522Marlar-Pavey MShen QStein MCFiolka RLin HYSturm GFeng BGalecki SLi ALGillich AMillett-Sikking ATang QChen YZhou FDaetwyler SMonistrol JJenkins EChang BJShalizi ADiaz UChen BJiou JMarshall WFFriedman JRTomer RShahmoradian SDean KMfalseMulti-immersion Oblique Plane Microscope (miOPM): A reconfigurable platform for high-resolution Light-Sheet Fluorescence Microscopy.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.2025-01-01T00:00:00Z2025 Oct2026-06-10T03:13:13.992Z2026-06-10T03:07:46.052ZS-EPMC126325404127904010.1101/2025.10.04.680473