Abstract
We present a modular implementation of structured illumination microscopy (SIM) that is fast, largely self-contained and that can be added onto existing fluorescence microscopes. Our strategy, which we call HIT-SIM, can theoretically deliver well over 50 super-resolved images per second and is readily compatible with existing acquisition software packages. We provide a full technical package consisting of schematics, a list of components and an alignment scheme that provides detailed specifications and assembly instructions. We illustrate the performance of the instrument by imaging optically large samples containing sequence-specifically stained DNA fragments.
Highlights
Fluorescence microscopy is a proven tool to study molecules, organelles, cells and whole organisms within their native context
The system consists of a conventional wide-field microscope that can generate periodic illumination patterns using a spatial light modulator (SLM)
Light from the laser source is directed onto the SLM, which acts as a configurable diffraction grating that can switch between patterns in less than 500 μs
Summary
Fluorescence microscopy is a proven tool to study molecules, organelles, cells and whole organisms within their native context. Two-dimensional SIM requires nine fluorescence images per reconstruction, allowing it to deliver fast imaging that is theoretically compatible with any type of labeling [1]. The speed and label-compatibility of SIM are major advantages over slower SR techniques such as PALM/STORM/SOFI [8,9,10,11], though the latter techniques may offer considerably higher spatial resolutions. These advantages make SIM especially suited to investigations that require information on systems that are highly dynamic [12,13,14]
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