Fig. 2

Patch2MAP enables super-resolution morphological and multiplexed proteomic imaging of human synapses. a, Two-photon z-stack and example somatic voltages in response to step current injections of a human L2/3 PC filled via somatic patch pipette with Alexa-488 and biocytin in an acute slice. b, Confocal z-stack of the same neuron at the intermediate expansion stage. Streptavidin Alexa-488 is used to show biocytin intracellular fill. C, Confocal images after re-slicing of the processed human brain slice into 4 thinner slices that contain the neuron shown in b. d, Confocal z-stacks of 3x expanded dendritic branch highlighted in a, b, c. e, Example basal branch from a different human neuron. Dendritic spine of interest indicated by green box. f, Magnified view of dendritic spine from e. g, Multi-round staining images of a Patch2MAP processed human tissue. Biocytin channels from both rounds were used as the reference for image registration. Round 1 (top): Bassoon (blue), GluN1 (yellow), and GluA1 & GluA2 (red). Round 2 (bottom): Shank3 (green), α-synuclein (magenta), and PSD-95 (cyan). Scale bars correspond to the physical dimensions of the tissue at the time of the imaging without accounting for the level of tissue expansion.