摘要：SummaryOptical visualization of complex microstructures in the entire organ is essential for biomedical research. However, the existing methods fail to accurately acquire the detailed microstructures of whole organs with good morphological and biochemical preservation. This study proposes a cryo-fluorescence micro-optical sectioning tomography (cryo-fMOST) to image whole organs in three dimensions (3D) with submicron resolution. The system comprises a line-illumination microscope module, cryo-microtome, three-stage refrigeration module, and heat insulation device. To demonstrate the imaging capacity and wide applicability of the system, we imaged and reconstructed various organs of mice in 3D, including the healthy tongue, kidney, and brain, as well as the infarcted heart. More importantly, imaged brain slices were performed sugar phosphates determination and fluorescencein situhybridization imaging to verify the compatibility of multi-omics measurements. The results demonstrated that cryo-fMOST is capable of acquiring high-resolution morphological details of various whole organs and may be potentially useful for spatial multi-omics.Graphical abstractDisplay OmittedHighlights•Cryo-fluorescence micro-optical sectioning tomography (Cryo-fMOST) was achieved•3D imaging of whole mouse tongue, kidney, heart, and brain at submicron resolution•Frozen state well preserves tissues’ original morphology and biochemical information•Cryo-fMOST is compatible with sugar phosphates determination and FISH measurementBiological sciences; Natural sciences; Neuroscience; Techniques in neuroscience;
