Fig. 5: Revealing 21 proteins on primary hippocampal neurons by ChemiPlex. | Nature Communications

Fig. 5: Revealing 21 proteins on primary hippocampal neurons by ChemiPlex.

From: NanoPlex: a universal strategy for fluorescence microscopy multiplexing using nanobodies with erasable signals

Fig. 5: Revealing 21 proteins on primary hippocampal neurons by ChemiPlex.

a Volumetric confocal image acquired after 7 repetitive cycles of ChemiPlex using 3-channel per cycle. b Categories of imaged targets comprised of 5 filaments, 3 organelles, 7 neuronal proteins, and 6 pre- and post-synaptic proteins. c High magnification images from individual proteins acquired during 21-ChemiPlex. From cycle 1 to 7 the following targets were revealed: Gephyrin (Geph), Synaptotagmin 1 (Syt1), Neuroligin 2 (NRL2), alpha-Synuclein (a-Syn), Rab3a, PMP70, beta-Spectrin (bSpectrin), Homer, Bassoon, Microtubule Associated Protein 2 (MAP2), Synaptic Vesicle protein 2b (SV2b), SNARE Associated Protein 25(SNAP25), vGAT, Vimentin, Doc2a, Clathrin, alpha-Tubulin, Synaptobrevin 2 (VAMP2), Post Synaptic Density 95 (PSD-95), vGluT1 and GFAP. Atto643, AZDye568, and Atto488 were used for channels 1, 2, and 3, respectively. Details on antibodies and one-step IF are in Methods and Supplementary Table 8.

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