Supplementary Figure 9: 3D super-resolution reconstruction and soSPIM versus wide-field single-molecule detection.
From: 3D high- and super-resolution imaging using single-objective SPIM
(a) Super-resolution lateral (xy) and orthogonal (xz and yz) reconstructions of the nucleus membrane protein Lamin-B1 labeled with the photoactivatable dye FLIP 565 in suspended S180 cells within a 24 × 24 µm2 microwell. Orthogonal reconstructions were performed from 14 planes (Fig. 3) acquired every 540 nm at between 6 and 13 µm above the coverslip. Scale bar, 5 µm. (b) Example of one of the 4,000 frames acquired during a STORM sequence acquisition on the nucleus membrane protein Lamin-B1 labeled with Alexa Fluor 647 dyes in a suspended S180 cell within a 24 × 24 µm2 microwell. The image on the left was acquired with a 1.5-µm-thick light sheet using soSPIM. The image on the right was acquired using a wide-field illumination performed with a collimated laser beam centered on the microwell. The graph shows the line intensity profiles of two single-molecule events acquired with soSPIM illumination (red line) and wide-field illumination (blue line). Scale bar, 1 µm.
