eSRRF supplementary data The eSRRF preprint is available here: https://doi.org/10.1101/2022.04.07.487490 Find the code and documentation here: https://github.com/HenriquesLab/NanoJ-eSRRF 1a: Set01a_DNA-PAINT_COS7-MT.tif DNA-PAINT of fixed COS-7 cells with indirect immunolabeling against tubulin Pixelsize: 160 nm Exposure time: 30 ms 50 000 frames 1b: Set01b_DNA-PAINT_COS7-MT_bin100.tif Dataset 1a with 100 frames binning to effectivly increase emitter density. eSRRF analysis parameters: M=5, R=0.5, S=1 all frames 2: Set02_HeLa_ffDronpa-MAP4.tif Live-cell TIRF imaging dataset of HeLa cells expressing ffDronpa-MAP4 Pixelsize: 156 nm Exposure time: 40 ms eSRRF analysis parameters: M=4 R=2 S=4 100 frames rolling analysis gap = 50 frames 3: Set03_COS7_PrSS-mEmerald-KDEL.tif Live-cell HiLO-TIRF of COS-7 cells expressing PrSS-mEmerald-KDEL marking the ER lumen. Pixelsize: 160 nm Exposure time: 5 ms eSRRF analysis parameters: M=5 R=2 S=1 100 fr rolling analysis gap = 10 frames 4a: Set4a_MFM_HeLa_TOM20.tif Live-cell MFM of HeLa cells expressing TOM20-HaloTag labeled with JF549 Pixelsize: 160 nm Exposure time: 20 ms eSRRF analysis parameters: M=4 R=2 S=1 100 fr rolling analysis gap = 25 frames 3D axial offset = 390 nm 4b: Set4b_MFM_beadcalibration.tif MFM calibration data with TetraSpeck beads immobilized on a coverslip, Pixelsize: 160 nm z-step 60 nm