2019
DOI: 10.1101/857516
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Multi-colour DNA-qPAINT reveals how Csk nano-clusters regulate T-cell receptor signalling

Abstract: Phosphorylation of the negative regulatory element of the tyrosine kinase Lck by Csk downmodulates T-cell receptor induced signalling. Being constitutively active, Csk spatial organization is responsible for regulating this signalling interaction. Here, we used stoichiometrically accurate, multiplexed, single-molecule super-resolution microscopy (DNA-qPAINT) to image the nanoscale spatial architecture of Csk and two binding partners implicated in its membrane association -PAG and TRAF3. Combined with a newly d… Show more

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“…The high level of nanometre accuracy attained by DNA-PAINT (i.e. it can routinely obtain sub-10 nm localization precision) has proven capable of imaging individual molecular targets on synthetic samples that simulates biomolecular nanoclusters [18], as well as protein clusters in fixed cells [19][20][21]. Furthermore, due to the predictable binding kinetics between imager and docking strands, it is possible to correlate the frequency of single-molecule events with the underlying number of labelled molecular targets [22] overcoming 'overcounting' artefacts observed with other SMLM techniques.…”
Section: Introductionmentioning
confidence: 99%
“…The high level of nanometre accuracy attained by DNA-PAINT (i.e. it can routinely obtain sub-10 nm localization precision) has proven capable of imaging individual molecular targets on synthetic samples that simulates biomolecular nanoclusters [18], as well as protein clusters in fixed cells [19][20][21]. Furthermore, due to the predictable binding kinetics between imager and docking strands, it is possible to correlate the frequency of single-molecule events with the underlying number of labelled molecular targets [22] overcoming 'overcounting' artefacts observed with other SMLM techniques.…”
Section: Introductionmentioning
confidence: 99%