Dual-scale and dual-channel oblique plane microscopy for multi-resolution imaging of large tissue specimens

Oblique plane microscopy (OPM) is a single-objective implementation of light sheet fluorescence microscopy (LSFM) that enables volumetric imaging in an open-top configuration suitable for large tissue specimens. Such applications require both detailed visualization of cellular structures and rapid coverage of large areas. However, achieving these two capabilities simultaneously within a single OPM system remains challenging. Here, we present a dual-scale and dual-channel OPM platform (D²OPM) that alternates between two primary objectives mounted on a turret to enable micro-scale high-resolution imaging and meso-scale high-throughput imaging within a unified optical architecture. To maintain consistent remote-focus performance during objective switching, the illumination geometry is adjusted without introducing additional imaging arms. The micro-scale mode provides submicron lateral resolution for cellular assessment, while the meso-scale mode enables rapid centimeter-scale image acquisition. Because high-resolution volumetric imaging inherently limits acquisition speed, controlled under-sampling is applied in the micro-scale mode and combined with deep learning-based image restoration to preserve morphologic detail while increasing throughput. Dual-channel fluorescence imaging is achieved using dual-wavelength excitation and spectral separation onto a single camera. Imaging of human colon, stomach, and pancreatic specimens demonstrates structural and cytologic features comparable to hematoxylin and eosin histology, with quantitative agreement in nucleus size measurements. These results establish D²OPM as a compact multi-resolution LSFM platform for rapid histomorphologic assessment.