In the present study, we investigated the dynamics of a femtosecond (fs) laser induced bio-printing with cell-free and cell-laden jets under the variation of laser pulse energy and focus depth, by using time-resolved imaging. By increasing the laser pulse energy or decreasing the focus depth thresholds for a first and second jet are exceeded and more laser pulse energy is converted to kinetic jet energy. With increasing jet velocity, the jet behavior changes from a well-defined laminar jet, to a curved jet and further to an undesired splashing jet. We quantified the observed jet forms with the dimensionless hydrodynamic Weber and Rayleigh numbers and identified the Rayleigh breakup regime as the preferred process window for single cell bioprinting. Herein, the best spatial printing resolution of 42 ± 3 µm and single cell positioning precision of 12.4 µm are reached, which is less than one single cell diameter about 15 µm.
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