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. 2024 Aug 5;15:6648. doi: 10.1038/s41467-024-50411-z

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a 2D CLEM image showing the fluorescence signal (green) of the calbindin-specific scFv probe overlapping with the cell body of a Purkinje cell. b EM image showing 2D segmentation (green) of the calbindin-positive Purkinje cell (n = 1). c 3D reconstruction of the Purkinje cell labeled in a (n = 1), with the cell body in dark green and a dendritic branch in light green; three parallel fibers (red) make synapses on three spine heads of the dendritic branch (arrow indicates a parallel fiber (PF); arrowhead indicates a synapse). d EM image showing 2D segmentation of the synapse (arrowhead) between a parallel fiber (red) and a spine head of the dendritic branch (green) (n = 1). e 2D CLEM image showing fluorescence signals (red) of the GFAP-specific scFv probe overlapping with the cell body of a velate astrocyte in the granule cell layer (n = 1). f EM image showing 2D segmentation (red) of the velate astrocyte in (e) (n = 1). g 3D reconstruction of the velate astrocyte (red) labeled in (e) and two nearby granule cells (GC1 and GC2, light and dark blue) (n = 2); the astrocyte extends a veil-like glial process (arrowhead) between the two granule cells. h EM image showing 2D segmentation of the glial process (arrowhead) between GC1 and GC2 (n = 1). i 2D CLEM image showing fluorescence signals (red) of the GFAP-specific scFv probe overlapping with a Bergmann fiber (n = 1). j EM image showing 2D segmentation (red) of the Bergmann fiber in (i) (n = 1). k 3D reconstruction of two Bergmann glial cells (BG1 and BG2) (n = 2) traced from their Bergmann fibers labeled by the GFAP-specific scFv probe. l EM image showing 2D segmentation of the cell body of BG2 (n = 1) and a nearby basket cell (n = 1), noting the lack of infoldings in BG2’s nuclear membrane compared to the basket cell. n indicates an example.