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Focus image scanning microscopy for sharp and gentle super-resolved microscopy. | LitMetric

AI Article Synopsis

  • Super-resolution microscopy faces challenges in imaging live and thick samples due to high illumination intensity and out-of-focus background interference.
  • The authors propose a new method called STED-ISM, which utilizes a detector array to enhance typical STED microscopy, reducing the intensity needed for high-resolution imaging.
  • They also introduce Focus-ISM, an innovative strategy to improve optical sectioning and eliminate background noise, requiring only minimal changes to conventional microscopes while benefiting live sample imaging.

Article Abstract

To date, the feasibility of super-resolution microscopy for imaging live and thick samples is still limited. Stimulated emission depletion (STED) microscopy requires high-intensity illumination to achieve sub-diffraction resolution, potentially introducing photodamage to live specimens. Moreover, the out-of-focus background may degrade the signal stemming from the focal plane. Here, we propose a new method to mitigate these limitations without drawbacks. First, we enhance a STED microscope with a detector array, enabling image scanning microscopy (ISM). Therefore, we implement STED-ISM, a method that exploits the working principle of ISM to reduce the depletion intensity and achieve a target resolution. Later, we develop Focus-ISM, a strategy to improve the optical sectioning and remove the background of any ISM-based imaging technique, with or without a STED beam. The proposed approach requires minimal architectural changes to a conventional microscope but provides substantial advantages for live and thick sample imaging.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9747786PMC
http://dx.doi.org/10.1038/s41467-022-35333-yDOI Listing

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