Robust fluctuation-based super-resolution microscopy in a confocal architecture
arxiv(2023)
摘要
Record-holding super-resolution microscopy (SRM) techniques, e.g.
localization microscopy (LM) and stimulated-emission-depletion (STED)
microscopy, require significant resources and effort from life-science
researchers. In comparison with confocal microscopy, apart from relying on
extensive sample-staining procedures and long acquisition times, applying LM
for 3D and multicolor imaging poses a considerable experimental challenge. In
the current work, we provide a complete demonstration of an entry-level SRM
technique - providing super-resolving capabilities with an experimental
complexity level akin to that of confocal microscopy. Exchanging the confocal
pinhole with a small pixelated detector, we use the inherent fluctuations of
dye molecules as contrast for SRM. This contrast is processed into a
super-resolved image through a process of pixel reassignment, a robust and
deterministic algorithm. Since the fluctuation contrast is ubiquitous to
organic markers, it does not require engineering of the blinking statistics
through the sample buffer. Together with the built-in capabilities for
multi-color and 3D imaging, shown in our work, it can become a natural
extension to confocal microscopy - a straightforward first step into the realm
of SRM.
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