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We thank B. Hajj (Institut Curie), J. Chen (Janelia Farm Research Campus), I. Izeddin (Ecole Normale Supérieure), J. Sillibourne (Commissariat à l'Energie Atomique et aux Energies Alternatives) and M. Bornens (Institut Curie) for providing super-resolution data sets used in the figures of this correspondence. We are grateful to K. Busch (Universität Osnabrück) for sharing the plasmid TOM-20-GFP. We also thank N. Clack, F. Mueller and X. Darzacq for discussions during the preparation of this work. This work was supported by funding from the Transcription Imaging Consortium at the Janelia Farm Research Campus, the Institut Curie International PhD Program and by the Tridimic grant from the program Recherches Partenariales et Innovation Biomédicale from Agence Nationale pour la Recherche.
The authors declare no competing financial interests.
ViSP executables for Windows and Mac OS X (with User Manual and License) (ZIP 15640 kb)
Rotation of a 3D STORM acquired mammalian mitochondria segment (U2OS cell transfected with TOM20-GFP and nanobody label with Alexa 647) to illustrate differences in localization precision between the lateral and axial dimensions using ViSP. The left panel is a reconstruction with isotropic Gaussians, while the right panel is that of a reconstruction with anisotropic Gaussians, revealing a poorer localization precision in the axial (z) dimension. Size of region is 1.4 × 3.2 × 1.8 μm. (MOV 3790 kb)
Left panel shows a 200,000 localization reconstruction of a 3D STORM acquired mammalian mitochondria network (U2OS cell transfected with TOM20-GFP and nanobody labeling with Alexa 647). Right panel is a surface rendering and cluster segmentation of the mitochondria after a local density threshold using ViSP. (MOV 27813 kb)
Left panel is a localization-based representation of a multi-channel centrosome complex (mEos2-Centrin1 in red and Alexa 647-labelled Cep 164 in blue) in a combined 3D PALM/STORM acquisition. Right panel is the representation of the same channels using ViSP's surface rendering feature. (MOV 13957 kb)
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Beheiry, M., Dahan, M. ViSP: representing single-particle localizations in three dimensions. Nat Methods 10, 689–690 (2013). https://doi.org/10.1038/nmeth.2566
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