Microscope dmi3000 b

Bright-field and epifluorescence images of the vesicles were taken by a ×63 numerical aperture (NA) 1.3 oil immersion objective on a commercially built Leica Microscope DMI3000 B in combination with a Hamamatsu ORCA-ER camera. Confocal images were taken by a ×63 NA 1.4 oil immersion objective with on a Leica TSC SP5. The temporal evolution was observed by acquiring 3D stacks of epifluorescence snapshots with a ×100 NA 1.4 oil immersion objective or a ×40 NA (1.25–0.75) oil immersion objective on a commercially built Leica Microscope DMI3000 B. The 3D image acquisition was started one minute after finishing the encapsulation process. We acquired snapshots of the 3D epifluorescence stacks of the vesicles every three minutes in order to avoid any artefacts by phototoxicity.
Imaging of fixed cells was performed with an LSM510Meta confocal microscope (Zeiss) equipped with a ×63/1.3 Plan-Neofluar objective using the 488 nm and 543 nm laser lines.
Confocal and fluorescent images were used to measure the domain length and curvature by ImageJ. Actin domains at the membrane were counted with the 3D object counter provided by ImageJ.

Images of the webs were taken using a Leica M205C stereo microscope and a Leica Microscope DMI 3000B. For scanning electron microscopy (SEM), all samples were sputter-coated with platinum (2 nm coating) and imaged using a ZEISS Sigma VP 300.

Vesicles were imaged with a Leica Microscope DMI3000 B and a 63× numerical aperture 1.3 oil immersion objective for bright-field microscopy and epifluorescence, in combination with a Hamamatsu ORCA-ER camera (Hamamatsu, Japan).
Confocal images were acquired using Leica TSC SP5 and a 63× numerical aperture 1.4 oil immersion objective. The three-dimensional (3D) reconstruction using the confocal stack was done using Imaris Software. Kymographs were prepared from the time-lapse images of the vesicles sedimented on passivated surface in epifluorescence using a Fiji (27 (link)) plugin. We always used the closed chambers to image the vesicles to avoid large scale drifts and convection.