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Article Dans Une Revue Proceedings of the National Academy of Sciences of the United States of America Année : 2014

Fast high-resolution 3D total internal reflection fluorescence microscopy by incidence angle scanning and azimuthal averaging.

Résumé

Total internal reflection fluorescence microscopy (TIRFM) is the method of choice to visualize a variety of cellular processes in particular events localized near the plasma membrane of live adherent cells. This imaging technique not relying on particular fluorescent probes provides a high sectioning capability. It is, however, restricted to a single plane. We present here a method based on a versatile design enabling fast multiwavelength azimuthal averaging and incidence angles scanning to computationally reconstruct 3D images sequences. We achieve unprecedented 50-nm axial resolution over a range of 800 nm above the coverslip. We apply this imaging modality to obtain structural and dynamical information about 3D actin architectures. We also temporally decipher distinct Rab11a-dependent exocytosis events in 3D at a rate of seven stacks per second.

Dates et versions

hal-01133826 , version 1 (20-03-2015)

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Citer

Jérôme Boulanger, Charles Gueudry, Daniel Münch, Bertrand Cinquin, Perrine Paul-Gilloteaux, et al.. Fast high-resolution 3D total internal reflection fluorescence microscopy by incidence angle scanning and azimuthal averaging.. Proceedings of the National Academy of Sciences of the United States of America, 2014, 111 (48), pp.17164-9. ⟨hal-01133826⟩
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