Full volume super-resolution imaging of thick mitotic spindle using 3D AO STED microscope

David McGloin, Jason Swedlow (Lead / Corresponding author), Piotr Zdankowski

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)
150 Downloads (Pure)

Abstract

Stimulated emission depletion (STED) nanoscopy is one of a suite of modern optical microscopy techniques capable of bypassing the conventional diffraction limit in fluorescent imaging. STED makes use of a spiral phase mask to enable 2D super-resolution imaging whereas to achieve full volumetric 3D super-resolution an additional bottle-beam phase mask must be applied. The resolution achieved in biological samples 10 μm or thicker is limited by aberrations induced mainly by scattering due to refractive index heterogeneity in the sample. These aberrations impact the fidelity of both types of phase mask, and have limited the application of STED to thicker biological systems. Here we apply an automated adaptive optics solution to correct the performance of both STED masks, enhancing robustness and expanding the capabilities of this nanoscopic technique. Corroboration in terms of successful high-quality imaging of the full volume of a 15μm mitotic spindle with resolution of 50nm x 50nm x 150nm achieved in all three dimensions is presented.

Original languageEnglish
Article number#352466
Pages (from-to)1999-2009
Number of pages11
JournalBiomedical Optics Express
Volume10
Issue number4
Early online date25 Mar 2019
DOIs
Publication statusPublished - 1 Apr 2019

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