In-depth analysis of vapor nanobubble-mediated permeabilization at the level of the plasma membrane and nuclear envelope

Vapor nanobubble (VNB) photoporation efficiently permeabilizes a wide range of cell types, with minimal toxicity. Despite its promise, however, little is known about its effects on cell homeostasis. Elucidation of the cellular response to VNB-induced plasma membrane disruption could help to diminish side effects and thereby increase efficiency of intracellular delivery. In this thesis, we set out to investigate the cellular response to VNB photoporation. We find a mechanoresponse to occur at the level of the nucleus, as shown by the increase in A-type lamins and chromatin compaction. We find that the observed nuclear stiffening is required for the cell to cope with the effects of VNB photoporation. Finally, we show that the obtained knowledge can be exploited to increase the efficiency of VNB photoporation, which will be vital for the technique to compete with other intracellular delivery methods. In some applications, cytosolic delivery is not sufficient and introduction into the nucleus is required. Controlled disruption of the nuclear envelope (NE) could also help to study the consequences of NE ruptures (NERs), which have been found to occur in cells with a vulnerable NE. Detailed investigation of NER consequences is difficult because of their stochastic nature. Thus, a method that induces NERs with spatiotemporal control would allow to study their consequences in more detail. In this thesis, we investigate the potential of VNB photoporation for disrupting the NE. We show that NE photoporation allows to induce NERs that mimick the spontaneous NERs to a high degree. Moreover, we find that NE photoporation can facilitate delivery of molecules into the nucleus. Once fully optimized, NE photoporation will not only be a substantial asset for research on NERs, it will become a valuable tool for a wide range of applications that require the transient and controlled disruption of the NE