The lamellarity distribution of liposomes characterized with multipass resistive-pulse sensing and fluorescence imaging is well approximated by the lamellarity distribution obtained from the cryoTEM imaging and shown in Figure . Thus, we assign the data points within the green-shaded region of Figure to unilamellar liposomes and the data points within the blueshaded region to multi-vesicle liposomes. The assignment of liposome lamellarity in the resistive pulse studies is only approximate due to photobleaching and the error in the fluorescence signal. Total membrane surface area of all lamellar structures within a liposome vs. the outer surface area of the liposome and liposome radius. The total membrane surface area was calculated from the liposome radii measured in the cryoTEM images and includes the surface area of both sides of the phospholipid membrane(s) assuming a membrane thickness of 4.1 ± 0.3 nm. 406 liposomes with radii >50 nm were analyzed. The green points correspond to unilamellar liposomes, blue correspond to MV1 liposomes, and red corresponds to MV2 liposomes. (E) Average number of photons per translocation vs. the outer surface area of the liposome for 74 individual liposomes characterized with resistive-pulse sensing. The surface area of the liposome exterior was calculated using rL from 5 liposome multipasses. Based on the liposome lamellarity distribution observed in (D), the green shading corresponds to unilamellar liposomes while the blue shading corresponds to multi-vesicle liposomes. This lamellarity assignment in (E) is only approximate. The percent distribution obtained from the cryoTEM images of unilamellar and multi-vesicle liposomes with radii >50 nm was considered in the assignment.