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This lecture by the instructor covers the methodology of single-molecule localization microscopy, focusing on the transition from single-molecule localizations to single proteins maps. The presentation includes details on biological samples, DNA-PAINT, protein distribution, k-means clustering, and the application of these techniques in T-cell signaling. Pros and cons of the methodology are discussed, highlighting the limitations when counting localizations and the reliability of k-means clustering. The lecture concludes with acknowledgments to collaborators and funding sources.