Extended Data Fig. 5: Algorithm flowchart for 4Pi-SIMFLUX reconstruction.
From: 4Pi-SIMFLUX: 4Pi single-molecule localization microscopy with structured illumination
Refer to the Methods section and Supplementary Note 2 for a detailed description. Rough XY ① and Z positions ② were estimated by MLE fitting with an elliptical Gaussian PSF model. 4Pi-SIMFLUX integrates SIMFLUX’s precise XY positions ③ and 4Pi-SMLM’s precise Z position ④ to obtain precise XYZ positions ⑤. In filtering step 1, molecules that are not fluorescent in all six frames were rejected using two filters: photon asymmetry ratio and modulation depth. In filtering step 2, molecules were rejected using other filters such as photon number, localization precision and interference contrast. Please see Supplementary Table 2 and Supplementary Note 2 for details.
