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Three-dimensional nanometre localization of nanoparticles to enhance super-resolution microscopy.

Bon P, Bourg N, Lécart S, Monneret S, Fort E, Wenger J, Lévêque-Fort S - Nat Commun (2015)

Bottom Line: Here we demonstrate fast full three-dimensional nanometre super-localization of gold nanoparticles through simultaneous intensity and phase imaging with a wavefront-sensing camera based on quadriwave lateral shearing interferometry.We show how to combine the intensity and phase information to provide the key to the third axial dimension.We demonstrate that nanoscale stabilization greatly enhances the image quality and resolution in direct stochastic optical reconstruction microscopy imaging.

View Article: PubMed Central - PubMed

Affiliation: 1] Laboratoire Photonique Numérique et Nanosciences (LP2N), CNRS UMR5298, Institut d'Optique Graduate School, Bordeaux University, Rue Francois Mitterand, 33400 Talence, France [2] Institut Langevin, ESPCI ParisTech, CNRS UMR 7587, PSL Research University, 1 rue Jussieu, Paris 75238, France [3] Institut des Sciences Moléculaires d'Orsay (ISMO), University Paris-Sud, CNRS UMR 8214, Orsay 91405, France.

ABSTRACT
Meeting the nanometre resolution promised by super-resolution microscopy techniques (pointillist: PALM, STORM, scanning: STED) requires stabilizing the sample drifts in real time during the whole acquisition process. Metal nanoparticles are excellent probes to track the lateral drifts as they provide crisp and photostable information. However, achieving nanometre axial super-localization is still a major challenge, as diffraction imposes large depths-of-fields. Here we demonstrate fast full three-dimensional nanometre super-localization of gold nanoparticles through simultaneous intensity and phase imaging with a wavefront-sensing camera based on quadriwave lateral shearing interferometry. We show how to combine the intensity and phase information to provide the key to the third axial dimension. Presently, we demonstrate even in the occurrence of large three-dimensional fluctuations of several microns, unprecedented sub-nanometre localization accuracies down to 0.7 nm in lateral and 2.7 nm in axial directions at 50 frames per second. We demonstrate that nanoscale stabilization greatly enhances the image quality and resolution in direct stochastic optical reconstruction microscopy imaging.

No MeSH data available.


Related in: MedlinePlus

Nanometre super-localization accuracy of a 100 nm gold nanoparticle.(a) Scatter plot of the nanoparticle lateral x and axial z positions showing 400 measurements at 50 frames per second video rate (20 ms acquisition time per measurement, the numerical computation time is negligible). The x and z position histograms are represented in grey. (b) s.d. of the measured nanoparticle position along x (black line) and z (red dashed line) as a function of the calibrated axial defocus induced by the piezo stage. For each axial stage position, 100 measurements are taken at 50 Hz to estimate the s.d. of the localization. The noise level corresponding to the set-up vibrations over the 2 s integration time is represented by a grey dashed line for the x direction and by an orange dashed dot line for the z direction. (c) Measured axial position as a function of the calibrated piezo stage z displacement.
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f2: Nanometre super-localization accuracy of a 100 nm gold nanoparticle.(a) Scatter plot of the nanoparticle lateral x and axial z positions showing 400 measurements at 50 frames per second video rate (20 ms acquisition time per measurement, the numerical computation time is negligible). The x and z position histograms are represented in grey. (b) s.d. of the measured nanoparticle position along x (black line) and z (red dashed line) as a function of the calibrated axial defocus induced by the piezo stage. For each axial stage position, 100 measurements are taken at 50 Hz to estimate the s.d. of the localization. The noise level corresponding to the set-up vibrations over the 2 s integration time is represented by a grey dashed line for the x direction and by an orange dashed dot line for the z direction. (c) Measured axial position as a function of the calibrated piezo stage z displacement.

Mentions: We quantify the accuracy of our 3D super-localization technique using 400 repeated position measurements at 50 Hz video rate (20 ms acquisition plus processing time per measurement). Figure 2a displays a scatter plot of the lateral x and axial z positions. The standard deviation (s.d.) of the position measurement at 50 Hz acquisition rate is σ(pxy)=1.5 nm in the lateral directions and σ(pz)=6.5 nm in the axial direction. This corresponds to remarkable super-localization accuracies of ∼λ/400 along xy and ∼λ/90 along z direction.


Three-dimensional nanometre localization of nanoparticles to enhance super-resolution microscopy.

Bon P, Bourg N, Lécart S, Monneret S, Fort E, Wenger J, Lévêque-Fort S - Nat Commun (2015)

Nanometre super-localization accuracy of a 100 nm gold nanoparticle.(a) Scatter plot of the nanoparticle lateral x and axial z positions showing 400 measurements at 50 frames per second video rate (20 ms acquisition time per measurement, the numerical computation time is negligible). The x and z position histograms are represented in grey. (b) s.d. of the measured nanoparticle position along x (black line) and z (red dashed line) as a function of the calibrated axial defocus induced by the piezo stage. For each axial stage position, 100 measurements are taken at 50 Hz to estimate the s.d. of the localization. The noise level corresponding to the set-up vibrations over the 2 s integration time is represented by a grey dashed line for the x direction and by an orange dashed dot line for the z direction. (c) Measured axial position as a function of the calibrated piezo stage z displacement.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525210&req=5

f2: Nanometre super-localization accuracy of a 100 nm gold nanoparticle.(a) Scatter plot of the nanoparticle lateral x and axial z positions showing 400 measurements at 50 frames per second video rate (20 ms acquisition time per measurement, the numerical computation time is negligible). The x and z position histograms are represented in grey. (b) s.d. of the measured nanoparticle position along x (black line) and z (red dashed line) as a function of the calibrated axial defocus induced by the piezo stage. For each axial stage position, 100 measurements are taken at 50 Hz to estimate the s.d. of the localization. The noise level corresponding to the set-up vibrations over the 2 s integration time is represented by a grey dashed line for the x direction and by an orange dashed dot line for the z direction. (c) Measured axial position as a function of the calibrated piezo stage z displacement.
Mentions: We quantify the accuracy of our 3D super-localization technique using 400 repeated position measurements at 50 Hz video rate (20 ms acquisition plus processing time per measurement). Figure 2a displays a scatter plot of the lateral x and axial z positions. The standard deviation (s.d.) of the position measurement at 50 Hz acquisition rate is σ(pxy)=1.5 nm in the lateral directions and σ(pz)=6.5 nm in the axial direction. This corresponds to remarkable super-localization accuracies of ∼λ/400 along xy and ∼λ/90 along z direction.

Bottom Line: Here we demonstrate fast full three-dimensional nanometre super-localization of gold nanoparticles through simultaneous intensity and phase imaging with a wavefront-sensing camera based on quadriwave lateral shearing interferometry.We show how to combine the intensity and phase information to provide the key to the third axial dimension.We demonstrate that nanoscale stabilization greatly enhances the image quality and resolution in direct stochastic optical reconstruction microscopy imaging.

View Article: PubMed Central - PubMed

Affiliation: 1] Laboratoire Photonique Numérique et Nanosciences (LP2N), CNRS UMR5298, Institut d'Optique Graduate School, Bordeaux University, Rue Francois Mitterand, 33400 Talence, France [2] Institut Langevin, ESPCI ParisTech, CNRS UMR 7587, PSL Research University, 1 rue Jussieu, Paris 75238, France [3] Institut des Sciences Moléculaires d'Orsay (ISMO), University Paris-Sud, CNRS UMR 8214, Orsay 91405, France.

ABSTRACT
Meeting the nanometre resolution promised by super-resolution microscopy techniques (pointillist: PALM, STORM, scanning: STED) requires stabilizing the sample drifts in real time during the whole acquisition process. Metal nanoparticles are excellent probes to track the lateral drifts as they provide crisp and photostable information. However, achieving nanometre axial super-localization is still a major challenge, as diffraction imposes large depths-of-fields. Here we demonstrate fast full three-dimensional nanometre super-localization of gold nanoparticles through simultaneous intensity and phase imaging with a wavefront-sensing camera based on quadriwave lateral shearing interferometry. We show how to combine the intensity and phase information to provide the key to the third axial dimension. Presently, we demonstrate even in the occurrence of large three-dimensional fluctuations of several microns, unprecedented sub-nanometre localization accuracies down to 0.7 nm in lateral and 2.7 nm in axial directions at 50 frames per second. We demonstrate that nanoscale stabilization greatly enhances the image quality and resolution in direct stochastic optical reconstruction microscopy imaging.

No MeSH data available.


Related in: MedlinePlus