Confocal image of CA1 neuropil in organotypic hippocampal slice culture, with PSD95-HaloTag expressed via a genetic knock-in ( PSD95-HaloTag mouse line), labeling all excitatory postsynapses (orange) and virus-assisted delivery of synaptophysin 1-eGFP (SYP1, blue), highlighting a subset of presynaptic terminals. Denoising (Noise2Void 51 ) applied (top). Scale bar, 2 m. Overlay with isotropically super-resolved volumetric LIONESS data (middle and bottom). Orthogonal planes in xy and xz directions represented as maximum intensity projections spanning 150 nm, with positions of corresponding planes indicated by arrowheads at image edges. Confocal SYP1 and PSD95 signals extend beyond the 3D super-resolved cellular structures defined by LIONESS. Data are representative for n = 4 biological replicates.
