Multifluorescence High‐Resolution Episcopic Microscopy for 3D Imaging of Adult Murine Organs

• DOI: 10.1002/adpr.202100110
• Publication Date: 2021-05-26T20:04:58Z
• Authors: Claire Walsh; Natalie A. Holroyd; Eoin Finnerty; Sean G. Ryan; Paul W. Sweeney; Rebecca J. Shipley; Simon Walker-Samuel
• Keywords: Optical sectioning; Biological specimen

3D microscopy of large biological samples (>0.5 cm 3 ) is transforming research. Many existing techniques require trade‐offs between image resolution, sample size, and method complexity. A simple robust instrument with the potential to conduct large‐volume imaging currently exists in form optical high‐resolution episcopic (HREM). However, development date limited single‐fluorescent wavelength nonspecific eosin staining. Herein, developments realize HREM become multifluorescent (MF‐HREM) are presented. MF‐HREM a serial‐sectioning block‐facing wide‐field fluorescence technique, which does not tissue clearing or sectioning. Multiple detailed preparation postprocessing enable multiple specific stains show how these segmentation quantification data. The application demonstrated variety contexts: whole tumor vascular networks cell invasion xenograft tumors up 7.5 mm at resolutions 2.75 μm, glomeruli volume adult mouse kidney, white‐matter track orientation brain.