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PLA is an antibody-based, ultra-sensitive in situ technique for detecting protein-protein interactions. Two target proteins are recognized by specific primary antibodies, followed by binding of secondary antibodies conjugated with DNA oligonucleotide probes. When the two targets are within <40 nm, the probes are ligated into a circular template by a DNA ligase. Rolling circle amplification (RCA) generates long DNA products, which are detected by fluorescent oligonucleotide hybridization, with each interaction event appearing as a countable, discrete fluorescent focus.