Eremchev, M.Roesel, D.Dansette, P. -m.Michailovas, A.Roke, S.2023-07-172023-07-172023-07-172023-05-0110.1116/6.0002640https://infoscience.epfl.ch/handle/20.500.14299/199143WOS:001004430500001Cell-sized giant unilamellar vesicles (GUVs) are an ideal tool for understanding lipid membrane structure and properties. Label-free spatiotemporal images of their membrane potential and structure would greatly aid the quantitative understanding of membrane properties. In principle, second harmonic imaging is a great tool to do so, but the low degree of spatial anisotropy that arises from a single membrane limits its application. Here, we advance the use of wide-field high throughput SH imaging by SH imaging with the use of ultrashort laser pulses. We achieve a throughput improvement of 78% of the maximum theoretical value and demonstrate subsecond image acquisition times. We show how the interfacial water intensity can be converted into a quantitative membrane potential map. Finally, for GUV imaging, we compare this type of nonresonant SH imaging to resonant SH imaging and two photon imaging using fluorophores.BiophysicsMaterials Science, BiomaterialsMaterials Sciencemultiphoton microscopylabel-freerepetition-rategenerationneuronswatertext::journal::journal article::research article