Full volume super-resolution imaging of thick mitotic spindle using 3D AO STED microscope
Piotr Zdańkowski , David McGloin , Jason R. Swedlow
AbstractStimulated 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.
|Journal series||Biomedical Optics Express, ISSN 2156-7085|
|Publication size in sheets||0.5|
|Keywords in English||microscopy, super-resolution microscopy, fluorescence microscopy, adaptive optics, aberration correction|
|Score||= 140.0, 08-06-2020, ArticleFromJournal|
|Publication indicators||= 6; = 0; : 2018 = 1.565; : 2018 = 3.91 (2) - 2018=3.757 (5)|
|Citation count*||8 (2020-07-20)|
* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.