Fig. 2
From: A peptide tag-specific nanobody enables high-quality labeling for dSTORM imaging
Super-resolution imaging and analysis of differently labeled vimentin constructs. a Schematic illustration of labeling strategies used for comparative SRM imaging of native or ectopically expressed vimentin. b Representative PALM/dSTORM images of chemically fixed Hela cells expressing the corresponding constructs outlined in a or native vimentin (left panel). Insets show magnifications of representative vimentin filaments of varying thickness (1—thick, 2—medium, 3—thin, peripheral). Scale bars, 5 µm in main images, 1 µm in insets. c Filament widths as histograms (left) with a bin size of 75 nm (x-axis) plotted against relative fraction (y-axis). Full data are represented underneath the histograms as box + scatter plots with the same x-axis. The box marks the three quartiles and the whiskers mark 95% of all the data. The average lengthwise fluorophore coverage was calculated for each bin and plotted (right) as mean filament width (black line) and standard deviation (colored area) against relative fraction covered by fluorophores (y-axis). Width and lengthwise fluorophore coverage were analyzed for a total of 676 (bivVB6-NbAF647), 295 (PAmCherry), 724 (GFP-NbAF647), and 620 (bivBC2-NbAF647) filaments, N = 5 cells for each condition, cells, and selected filaments are shown in Supplementary Fig. 4. Image reconstruction details are given in Methods section
