Correlation-based nanometric localization using lattice SIM2 and dSTORM in PML nuclear bodies validated by calibration spheres

We evaluated the Lattice di-SIM 3D structural illumination method with a focus on Promyelocytic Leukemia Nuclear Bodies (PML NBs). Lattice SIM 2 's performance was compared to its predecessor using biological samples and calibration beads. Optimized SIM 2 parameters revealed PML NBs with a distinct ring-like morphology. Furthermore, we compared Lattice-SIM 2 with dSTORM, finding a strong correlation between blinking events and the SIM 2 mask. For 2D nanoscopic correlation, we used the long-lived imaging buffer Eternity, while the modified variant, Eternity-SIM, with a higher refractive index, provided improved 3D correlation for SIM 2 imaging. • Optimized Lattice SIM 2 parameters to reveal the ring-like circular morphology of Promyelocytic Leukemia Nuclear Bodies (PML NBs). • Correlation of Lattice SIM 2 to Lattice SIM and to dSTORM imaging using biological samples and fluorescent spheres. • Investigated the effectiveness of Lattice SIM 2 to minimize Z-elongation compared to Lattice SIM. • Correlative Lattice 3D SIM 2 -STORM nanoscopy with Eternity-SIM buffer to minimize refractive index differences.