Extended Data Fig. 5: Molecular identification of cellular and subcellular structures.
From: Light-microscopy-based connectomic reconstruction of mammalian brain tissue
a, Single plane of a LICONN imaging volume in the hippocampal CA1 stratum radiatum, including the structural channel (grey) and immunolabelling for myelin basic protein (magenta) and the astrocytic gap-junction protein connexin-43 (cyan). Magnified views highlight structures expressing the respective molecular markers: (i) Immunolabelling for myelin basic protein identifies myelin sheaths, where the regions largely devoid of signal in the structural LICONN channel correspond to lipid-rich myelination. (ii) Connexin-43 indicates gap junctions. b, Single plane from LICONN imaging volume in hippocampus, showing the structural channel (grey) and immunolabelling for GFAP (orange) and connexin-43 (cyan). GFAP is an intermediate filament protein, commonly used as astrocyte marker. Note the GFAP signal in the immediate vicinity of the capillary traversing the image, and the cell nucleus on the right in the periphery of the blood vessel. Panels in the bottom show the immunolabelling and structural channels separately, as indicated by the box. c, Single plane from a LICONN imaging volume in the CA3 stratum lucidum, with additional immunolabelling for vimentin (orange, here highlighting the endothelial lining of a blood capillary) and the peroxisomal marker PMP70 (70 kDa peroxisomal membrane protein). Note that peroxisomes do not produce pronounced contrast in the structural (protein density) channel but can be identified by specific labelling. The displayed combinations of immunolabellings for MBP/Cnx-43 were technically replicated n = 1 time, RIM1/2 and VGLUT1 n = 2 times, GFAP/Cnx-43 n = 2 times, vimentin/PMP70 n = 1 time.
