A new microscopy technique in SciSignal allows researchers to visualize proteins in space and time without attaching bulky tags or fluorescent molecules, which can interfere with the protein of interest.
Although current microscopy-based studies of signaling molecules have provided insights into the dynamics of T cell activation, they are often hampered by the need to attach tags or fluorophores to the proteins of interest. Farrelldeveloped a superresolution microscopy–based technique called pPAINT through which they visualized the spatiotemporal distribution of the phosphotyrosine-binding SH2 domains of various signaling and adaptor molecules in activated T cells on lipid bilayers.
To overcome these limitations, we developed a technique to visualize the nanoscale binding locations of signaling proteins by taking advantage of their native interaction domains. Here, we demonstrated that pPAINT is a new, single-molecule localization microscopy technique and used it to investigate T cell signaling by visualizing the Src homology 2 domain, which is common in signaling molecules.
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