3D easySTED Nanoscopy Illumination Optics with Multiple Wave Plates for Simplified Beam Alignment

초록

Stimulated emission depletion (STED) nanoscopy is a widely used technique for investigating subcellular structures with nanoscale resolution. A persistent challenge in STED nanoscopy is the need for precise collinear alignment of excitation and depletion beams in the illumination path, which complicates the experimental setup. To address this, our study proposes an illumination optics design for 3D easySTED nanoscopy that simplifies beam alignment. The proposed system couples the excitation and depletion beams into a single-mode optical fiber, ensuring their propagation through a shared optical path. It includes multiple wave plates to manipulate the polarization states and a spatial light modulator to modulate the phases of the excitation and depletion beams so that they can shape the depletion beam into a doughnut-shaped focus while maintaining a Gaussian focus for the excitation beam. This design facilitates alignment and supports the maintenance of optimal focus. Simulation results demonstrate that the proposed approach achieves resolution comparable to that of conventional 3D STED nanoscopy.

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