Is laser repetition rate important for two-photon light sheet microscopy?

Abstract

We demonstrate the thermal advantages of using low repetition rate, high peak power lasers for imaging in two-photon light sheet microscopy using a Bessel light beam. We compare the use of two ultrashort pulsed lasers in such an imaging system: a high repetition rate source operating at 80 MHz and a low repetition rate source operating at 1 MHz. The low repetition rate laser requires approximately one order of magnitude lower average power than the high repetition rate source to yield the same fluorescent signal. These lasers are used to image Zebrafish larvae and record their heart rates. The data show heart rate values 30% in excess of the ground truth baseline value when imaged with the high repetition rate source due to deleterious heating, whereas the low repetition rate source yields data only a few percent above this ground truth value.