Efficient multiphoton microscopy with picosecond laser pulses

Multiphoton microscopes employ femtosecond lasers as light sources because the high peak power of the ultrashort pulse allows for multiphoton excitation of fluorescence in the examined sample. However, such short pulses are susceptible to broadening in a microscope's highly dispersive optical elements and require careful dispersion management, otherwise decreasing excitation efficiency. Here, we have developed a 10 nJ Yb:fiber picosecond laser with an integrated pulse picker unit and evaluated its performance in multiphoton microscopy. Our results show that performance comparable to femtosecond pulses can be obtained with picosecond pulses only by reducing the pulse repetition rate and that such pulses are significantly less prone to the effect of chromatic dispersion. These findings proved that the temporal pulse compression is not always efficient, and it can be omitted by using a smaller and easier-to-use all-fiber setup.