The photoacoustic effect is generated when a variable light interacts with a strongly light-absorbing material. In water, it may produce hot bubbles and shock waves that could affect the integrity of nearby cellular membranes, opening transient pores (photoporation). In this study, we have evaluated the effect of pulsed laser-irradiated carbon nanoparticles (cNP) on model membranes and on Chinese hamster ovary (CHO) cells. Fluorescence lifetime measurements of calcein-loaded liposomes support the notion that the photoacoustic effect causes transient openings in membranes, allowing diffusion fluxes driven by gradient concentrations. With CHO cells, we have shown that this effect can induce either intracellular delivery of calcein, or release of cellular compounds. The latter process has been recorded live with multiphoton excitation microscopy during pulsed infrared laser irradiation. Calcein loading and cell viability were assayed by flow cytometry, measuring necrotic cells as well as those in early apoptosis. To further assess long-term cell recovery after the rather harsh treatment, cells were reseeded and their behaviour recorded for 48 h. These extended studies on cell viability show that pulsed laser cNP photoporation may be considered an adequate intracellular delivery technique only if employed with soft irradiation conditions (below 50 mJ/cm 2). Biological membranes constitute the cell boundaries, isolating the inner cell medium from the external environment. They consist of a hydrophobic matrix, formed by an oriented double layer of phospholipids (and/or glycolipids) to which proteins are bound in different forms 1. This barrier prevents large molecules from entering cells in a passive way. Only by active mechanisms, like receptor-mediated endocytosis, can they reach the cytoplasm 2. This is the basis of specific or receptor-targeted nanoparticle technology 3,4. However, this process may be slow, and it often leads molecules ultimately into lysosomes, thus to enzymatic degradation 5. In order to promote the introduction of drugs or genetic material avoiding the endocytic pathway, methods for direct cytoplasmic delivery are necessary 6. Different physical forces have been employed to induce transient openings in cell membranes, which can close fast enough as to not jeopardize cell living conditions. Examples of such methods are microinjection 7 , ultrasound 8 , electroporation 9 or light-irradiation techniques 10. In the latter category, the so-called photoacoustic effect is generated when a variable light interacts with a strong light-absorbent material e.g. black carbon 11,12. In aqueous media this effect can be easily achieved by irradiating a carbon nanoparticle (cNP) suspension with a pulsed infrared (IR) laser. Light energy absorption heats cNP above 1000 °C 13,14. Vaporization of the surrounding water and generation of acoustic emissions can impact nearby cells thermally-by contact with the hot bubblesand mechanically-by fluid mechanical forces-to physically disrupting cell membranes with transient...