Table 1.
Reconstruction parameters.
Fig 1.
Comparison of the different existing 2D blind-SIM algorithms on a paxillin sample illuminated with a distorted grating.
a) 2D blind-SIM using simultaneous estimation of the object and illumination functions [16]. b) 2D blind-SIM using the proposed method with sequential estimation. For improved comparability, a conjugate gradient scheme was used here. c) 2D WF deconvolution. The scale bar is 1 μm.
Fig 2.
Results of 3D simulation with thick slice reconstruction.
a) Resolution test target placed in the focal plane of our simulated sample. 800nm out-of-focus was a π/2-rotated version of the same structure. b) 2D WF deconvolution. c) Focal slice of 3D WF deconvolution of the entire WF image stack. d)One of the 9 simulated SIM images. Here we simulate a two-beam SIM. e) 2D blind reconstruction of (d) containing out-of-focus light. f) Thick slice blind-SIM result, showing optical sectioning and high resolution. g-i) Simulation as in a-f) but with a distorted illumination pattern as depicted in g) (zoom). h)Reconstructed illumination function. i) Thick slice blind-SIM reconstruction of the object described in (a) but illuminated with the distorted pattern(g). Scale bar: 2.5 μm.
Fig 3.
Experimental results in an MCF-7 actin-labelled cell.
A 200×200 pixels region of interest was selected to keep the computational time small. a) 2D wide-field (WF) deconvolution. b) 2D blind-SIM reconstruction with higher resolution but no optical sectioning. c) 3D WF deconvolution. d) Thick slice blind-SIM result. The resolution is improved and the out-of-focus contribution removed. The green arrows indicate a pair of filaments that is removed because it originally stems from another plane. The processing time was 25 min with GPU vs. 330 min without GPU. e) and f) are the reconstructions of the original 3D data with the ZEN software (version 2010D). e) Plane that was selected. f) Two slices under slice e), i.e. 182 nm away. The sample was prepared by Michael Reuter and data acquired by Elen Tolstik on a commercial ELYRA-S.1 SIM microscope (Carl Zeiss Microimaging, Jena, Germany). Scale bar: 2 μm.