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Chemotaxis of immune cells in vivo is important for their development,differentiation and proper function.However,no method has been developed for real-time measurement of single-cell responses to a specific chemotactic stimulus in vivo.We have developed the first two-photon caged chemokine probe[1-2],which is able to manipulate immune cell at a chosen tissue.This protein probe is synthesized through an efficient one-pot total chemical strategy in multi-milligram scale.Using this system,we analyze murine T lymphocyte chemotaxis in vitro at the single cell level,and manipulate the cell motility in eripheral tissues and secondary lymphoid organs.This provides a promising chemical tools to investigate the immune cell biology in vivo.