1. Benchmarking the volume measurement using polystyrene beads of defined size.
Benchmarking the volume measurement using polystyrene beads of defined size. (A) Polystyrene beads (Tetraspeck; ThermoFisher) of defined diameter were diluted in PBS, allowed to sediment to the bottom of the dishes, and imaged using a 3D Cell Explorer microscope. The small colored boxes depict digital RI stain of beads in red on black background. The large boxes show full 3D visualization of acquired holographs. 3D stacks of the beads were digitally stained (voxel segmented) for an RI estimated to cover at least 95% of the bead volumes. Bars, 5 µm. (B) Comparison of volume quantification performed by voxel summation either by voxel segmentation and counting in STEVE software or by surface fitting in Imaris. Bead diameter was computed based on voxel counts and voxel size and compared to the nominal diameter provided by the manufacturer.
Artur Yakimovich et al. mSphere 2018; doi: /mSphereDirect