Sharpening emitter localization in front of a tuned mirror - NPC dataset

This is a depository for two single molecule localisation microscopy datasets of nuclear pore complex (NPC) structures for single particle averaging. The data was published in: 

Heil, H.S., Schreiber, B., Götz, R. et al. Sharpening emitter localisation in front of a tuned mirror. Light Sci Appl 7, 99 (2018). https://doi.org/10.1038/s41377-018-0104-z

Both datasets have two different levels of localisation precision as one is a conventional STORM experiment and the second a mirror-enhanced STORM experiment. A detailed description of the sample preparation and imaging conditions can be found in the related publication. In short the NPC structures are placed on the surface of a glas coverslip or nano-mirror coated coverslip by manual isolation and spreading of nuclear envelopes from xenopus laevis oocytes, fixed and stained by indirect immunolabeling. The primary antibody targets GP210, the secondary F(ab')2 fragment is conjugated with Alexa Fluor 647. 

In this depository I'm providing the raw images data, localisation data and super-resolved reconstruction  for the two experiments, as well as the localisation data and super-resolved reconstruction of single NPC rings. 

I'm also providing a MatLab script that allows to select single NPC positions in the super-resolved image and export the localization data of the single NPC ROI: P01_ImageAlignment_PickElements.m

Information about the dataset is also available  here: NPC Image Alignment Dataset_Info.pdf.

Image parameters: 102 nm pixel size, EM Gain 100, Photoelectrons per A/D count: 15.01

Column structure of the localisation text files: 

Id,Frame, x [nm], y [nm], sigma [nm], intensity [photon], offset [photon], bkgstd [photon], chi2, Uncertainty [nm], detections

Files: 

NPCData_glass_EPI.tif

-> NPC on glass coverslip, low power EPI illumination, widefield image, 20 ms exposure

NPCData_glass_STORM.tif

-> NPC on glass coverslip, high power EPI illumination, 5 ms exposure, 20000 frames

NPCData_glass_STORM_loc.csv

-> ThunderSTORM Localisation data of NPCData_glass_STORM.tif, parameters specified NPCData_glass_STORM_loc-protocol.txt

NPCData_glass_STORM_20xNormalizedGaussian.tif

-> 20x Nomalized Gaussian reconstruction of localization data from NPCData_glass_STORM.tif (ThunderSTORM), pixelsize 5.1 nm

NPCData_glass_STORM_singleRings.zip

-> Localisation data and 20x 20x Nomalized Gaussian reconstruction of single NPC ROIs picked out of the NPCData_glass_STORM dataset, ROI size is 240*240 nm2

NPCData_nanomirror_EPI.tif

-> NPC on nanomirror coated coverslip, low power EPI illumination, widefield image, 20 ms exposure

NPCData_nanomirror_STORM.tif

-> NPC on nanomirror coated coverslip, high power EPI illumination, 5 ms exposure, 20000 frames

NPCData_nanomirror_STORM_loc.csv

-> ThunderSTORM Localisation data of NPCData_nanomirror_STORM.tif, parameters specified NPCData_nanomirror_STORM_loc-protocol.txt

NPCData_nanomirror_STORM_20xNormalizedGaussian.tif

-> 20x Nomalized Gaussian reconstruction of localisation data from NPCData_nanomirror_STORM.tif (ThunderSTORM), pixelsize 5.1 nm

NPCData_nanomirror_STORM_singleRings.zip

-> Localisation data and 20x 20x Nomalized Gaussian reconstruction of single NPC ROIs picked out of the NPCData_nanomirror_STORM dataset, ROI size is 240*240 nm2