An actively stabilized, miniaturized epi‐fluorescence widefield microscope for real‐time observation in vivo

Recent developments in real‐time, in vivo micro‐imaging have allowed for the visualization of tissue pathological changes, facilitating rapid diagnosis. However, miniaturization, magnification, the field of view, and in vivo image stabilization remain challenging factors to reconcile. A key issue for this technology is ensuring it is user friendly for surgeons, enabling them to use the device manually and obtain instantaneous information necessary for surgical decision‐making. This descriptive study introduces a handheld, actively stabilized, miniaturized epi‐fluorescence widefield microscope (MEW‐M) for real‐time observation in vivo with high resolution. The methodology of MEW‐M system includes high resolution microscopy miniaturization technology, thousandfold shaking suppression (actively stabilized), ultra‐photosensitivity, and tailored image signal processing cell image capture and processing technology, which support for the excellent real‐time imaging performance of MEW‐M system in brain, mammary, liver, lung, and kidney tissue imaging of rats in vivo. With a single‐objective and high‐frame‐rate imaging, the MEW‐M system facilitates roving image acquisition, enabling contiguous analysis of large tissue areas. Research Highlights A handheld, actively stabilized MEW‐M system was introduced. Excellent real‐time, in vivo imaging with high resolution and active stabilization in brain, mammary, liver, lung, and kidney tissue of rats. A newly designed anti‐shaking MEW‐M system, and the real‐time imaging of organ tissue in living rats versus H&E stained histological examination, including brain, mammary, liver, lung, and kidney.