Extended Data Fig. 1: Time step histograms.
From: Overlapping nuclear import and export paths unveiled by two-colour MINFLUX
a,b, The time between two successive MINFLUX localizations for NbGFP-HMSiR in datasets 1 and 2. The insets are EFO (emission frequency at offset) histograms. The EFO is a measure of the emission intensity. With a higher average EFO, the localization is less likely to fail since sufficient photons are collected within the dwell time of the localization. Longer localization times occur when the dwell time needs to be extended to collect enough photons. These EFO histograms reveal a significantly higher EFO peak within dataset 2 compared to dataset 1. This difference explains why the average localization time of dataset 1 was longer than dataset 2. We note that the different distributions of HMSiR localization times for the two datasets should have no material effect on the NPC reconstructions since the data were collected over 15–20 min and the NPC scaffolds have been shown to be stable over this time period7. Further explanation and discussion of the EFO can be found in the Methods and Extended Data Fig. 6. c,d, The time between two successive MINFLUX localizations for Imp α-JF549 in datasets 1 and 2. See Supplementary Tables 1 and 3 for acquisition parameters.
