Fig. 1

Patch2MAP enables super-resolution morphological and proteomic imaging in neural tissue without exogenous protein expression. a, Schematic of experimental tissue processing pipeline for full morphological labelling of individual neurons and subsequent cell-delineated super-resolution proteomic analysis. b, Left column: two-photon z-stack of a mouse L5 PC filled via somatic patch pipette with Alexa-488 and biocytin in an acute slice. Middle column: confocal z-stack of the same neuron at the stage of intermediate expansion. Strepatvidin Alexa-fluor 488 reveals biocytin. Right column: overlay of the two z-stacks. c, After intermediate expansion, the 300 μm-thick acute brain slice is resliced into 4 thinner slices. Confocal z-stacks show the 4 slices which contain the neuron from b. d, Confocal z-stacks of 3x-expanded soma and branches from the neuron in b and highlighted in c. e, Magnified view of basal branch from d. Green arrowheads indicate example spines shown to the right of the branch. From left to right: image of the cell-filling biocytin channel stained with Strepatvidin Alexa-fluor488, presynaptic protein Bassoon stained with anti-guinea pig-Alexa405 (blue), NMDAR subunit GluN1 stained with anti-mouse-Alexa555 (yellow), and AMPAR subunit GluA1 stained with anti-rabbit-Alexa647 (red). Scale bars correspond to the physical dimensions of the tissue at the time of the imaging without accounting for the level of tissue expansion.