Fig. 4

Comparison of the multi-color super-resolution experimental results on a fixed COS-7 cell sample with DAPI-labeled nucleus, Alexa Fluor^TM 568-labeled actin and MitoTracker^TM Green FM-labeled mitochondria. a Comparison of the wide-field image and the super-resolution images obtained by different methods (Open-3DSIM¹⁹ and PCA-3DSIM), where the top are lateral slices of layer 12 and the bottom are axial slices along the purple dashed line. The raw SIM images were captured through a 100\(\times\) objective (CFI Apochromat TIRF 100\(\times\) Oil, NA 1.49, Nikon, Japan) of N-SIM (Nikon, Japan). For easy distinguishing, we show the mitochondria, actin and nucleus of the COS-7 cell in green, red and blue, respectively. The mitochondria are excited at 488 nm wavelength, the nucleus at 405 nm wavelength and the actin at 561 nm wavelength. b–d Magnified wide-field image and super-resolution images from the white-, yellow-, and green-boxed regions in (a) obtained by different methods. e Intensity profiles along the blue lines in (b)–(d). f The lateral wave vector distribution of the tiled images along the x-direction in a specific layer and that of the tiled images at a particular lateral position in different layers. g rFRC maps of super-resolved images obtained by different methods for a certain layer (g1 and g2), as well as the distribution of rFRC values (g3), mean resolutions (g4), and contrasts (g5) for 3D super-resolution stack images obtained by different methods. h Slices of super-resolution image stacks obtained by PCA-3DSIM in different layers as well as the whole 3D sight. The experiments were independently repeated 10 times with similar results. Colored arrows point to regions where reconstruction differences are distinct. Scale bars: 3 μm (a); 1 μm (b, c); 0.5 μm (d); 5 μm (h). Scale on z-axis: 17 layers, 0.125 μm per layer