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. 2021 Dec 15;9:737621. doi: 10.3389/fcell.2021.737621

FIGURE 7.

FIGURE 7

Fluorescence images of immuno-labeled mouse cerebellum imaged with conventional FM and ILEM system (JEOL BSE detector). (A) An 80 µm slice of cerebellum labeled with calbindin D28 K primary and Alexa 488-conjugated secondary antibody before embedding with our IRF protocol. Imaged with conventional FM Nikon TiE inverted C2 confocal microscope Plan Apo 10x dry lens, labeled Purkinje cells can be easily recognized (asterisks). (B) After embedding with our IRF protocol, 150-nm cerebellar section imaged by conventional FM Nikon TiE inverted C2 confocal microscope Plan Apo 20x dry lens shows that the fluorescent labeling of Purkinje cells (asterisks) was preserved after embedding. (C) The same 150 nm section with a layer of Purkinje cells (asterisk, in B) imaged with fluorescent optics of ILEM-SEM (Plan Apo VC 100x lens) system shows a perfect correlation. Abbreviations: FM, fluorescence microscope; ILEM-SEM, integrated light and electron microscope; IRF, in-resin fluorescence; PC, Purkinje cell. Scale bars: A = 100 μm, B = 10 μm, C = 5 µm.