Аннотация:Owning to excellent optical properties and high biocompatibility carbon dots (CDs) have drawn increasing attention and have been widely applied as imaging agents for various bio-applications. Here we report a strategy for live-cell fluorescent bioimaging based on in situ synthesis of CDs within cells by tightly focused femtosecond laser pulses. Laser-produced carbon dots exhibit bright excitation-dependent fluorescence and are highly two-photon active under near infrared femtosecond excitation, thus demonstrating a potential for two-photon fluorescence imaging. The Raman spectra of fluorescent centers show strong D (1350 cm-1) and G (1590 cm-1) bands, thus suggesting that they are composed of carbon dots with sp2-hybridyzed core. Using Mouse GV oocytes as a model system we examine cytotoxicity and demonstrate the possibility of long-term fluorescent intracellular tracking of the laser-produced CDs. Created virtually in any point of the live cell, CD-based fluorescent μm-sized markers demonstrate high structural stability and retain bright fluorescence many hours after formation. Our results point to laser-produced fluorescent CDs as a highly-potent tool for cell cycle tracking, culture cell marking and probing intracellular movements.