Figure 3.

Impact of imaging conditions on data accuracy in PALM. (A) Representative images of iPSC-derived CM acquired with freshly prepared and used imaging buffer. Magnified sections (red frame) revealed differences in z-disc thickness when imaging conditions deteriorate. (B) Quantitative assessment demonstrated a significant difference in the thickness of sarcomere filaments, showing a much lower z-Disc size when imaging was performed with freshly prepared buffer (fresh vs. old buffer: 67.35 ± 1.05 nm vs. 116.6 ± 1.95 nm). (C) Low buffer quality reduces the number of detected photons and strongly impairs localization precision. Data are presented as mean ± SEM. 50 filaments have been subjected to analysis. Statistical significance was determined using the Student’s t-test. **** p < 0.0001. Scale bar 10 µm.