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Main Authors: Agazzi, Costanza, Toledo-García, Nick, Martín-Badosa, Estela, Montes-Usategui, Mario, Maluenda, David, Tiana-Alsina, Jordi, Martínez-Herrero, Rosario, Carnicer, Artur
Format: Preprint
Published: 2026
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Online Access:https://arxiv.org/abs/2603.27193
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author Agazzi, Costanza
Toledo-García, Nick
Martín-Badosa, Estela
Montes-Usategui, Mario
Maluenda, David
Tiana-Alsina, Jordi
Martínez-Herrero, Rosario
Carnicer, Artur
author_facet Agazzi, Costanza
Toledo-García, Nick
Martín-Badosa, Estela
Montes-Usategui, Mario
Maluenda, David
Tiana-Alsina, Jordi
Martínez-Herrero, Rosario
Carnicer, Artur
contents Fluorescence depletion microscopy techniques such as STED and RESOLFT require optical fields with a well-defined and spatially confined central intensity minimum to achieve sub-diffraction lateral resolution. Here, we present the design and experimental implementation of an azimuthally polarized, doughnut-shaped depletion beam based on super-pupil engineering principles. By tailoring the radial amplitude distribution at the entrance pupil to approximate a Bessel-type target function, the resulting focal field exhibits a tighter central doughnut compared to conventional azimuthally polarized beams. The designed pupil field distribution is implemented using a phase-only spatial light modulator operated in a double pass configuration, enabling independent modulation of orthogonal polarization components via complex-field holographic encoding. Experimental characterization using sub-diffraction fluorescent beads demonstrates a reduction of the peak-to-peak distance of the central doughnut by approximately 16% relative to a nominal azimuthally polarized reference beam. Although the engineered field exhibits pronounced sidelobes, these do not preclude its use as a depletion beam, since lateral resolution is strongly influenced by the spatial confinement and effective suppression of the central intensity minimum for a given depletion intensity. This suggests that the proposed approach can enable improved lateral resolution at comparable depletion powers, providing a flexible and experimentally accessible route for engineering depletion fields in reconfigurable super-resolution microscopy systems.
format Preprint
id arxiv_https___arxiv_org_abs_2603_27193
institution arXiv
publishDate 2026
record_format arxiv
spellingShingle Azimuthal super-pupil beam engineering for improved fluorescence depletion microscopy
Agazzi, Costanza
Toledo-García, Nick
Martín-Badosa, Estela
Montes-Usategui, Mario
Maluenda, David
Tiana-Alsina, Jordi
Martínez-Herrero, Rosario
Carnicer, Artur
Optics
Fluorescence depletion microscopy techniques such as STED and RESOLFT require optical fields with a well-defined and spatially confined central intensity minimum to achieve sub-diffraction lateral resolution. Here, we present the design and experimental implementation of an azimuthally polarized, doughnut-shaped depletion beam based on super-pupil engineering principles. By tailoring the radial amplitude distribution at the entrance pupil to approximate a Bessel-type target function, the resulting focal field exhibits a tighter central doughnut compared to conventional azimuthally polarized beams. The designed pupil field distribution is implemented using a phase-only spatial light modulator operated in a double pass configuration, enabling independent modulation of orthogonal polarization components via complex-field holographic encoding. Experimental characterization using sub-diffraction fluorescent beads demonstrates a reduction of the peak-to-peak distance of the central doughnut by approximately 16% relative to a nominal azimuthally polarized reference beam. Although the engineered field exhibits pronounced sidelobes, these do not preclude its use as a depletion beam, since lateral resolution is strongly influenced by the spatial confinement and effective suppression of the central intensity minimum for a given depletion intensity. This suggests that the proposed approach can enable improved lateral resolution at comparable depletion powers, providing a flexible and experimentally accessible route for engineering depletion fields in reconfigurable super-resolution microscopy systems.
title Azimuthal super-pupil beam engineering for improved fluorescence depletion microscopy
topic Optics
url https://arxiv.org/abs/2603.27193