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SOFISM: Super-resolution optical fluctuation image scanning microscopy
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DOI：10.1364/OPTICA.399600.png)
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Abstract
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Super-resolution optical microscopy is a rapidly evolving scientific field dedicated to imaging sub-wavelength-sized objects, leaving its mark in multiple branches of biology and technology. While several super-resolution optical microscopy methods have become a common tool in life science imaging, new methods, supported by cutting-edge technology, continue to emerge. One rather recent addition to the super-resolution toolbox, image scanning microscopy (ISM), achieves up to twofold lateral resolution enhancement in a robust and straightforward manner. To further enhance ISM's resolution in all three dimensions, we present and experimentally demonstrate here super-resolution optical fluctuation ISM (SOFISM). Measuring the fluorescence fluctuation contrast in an ISM architecture, we obtain images with a x2.5 lateral resolution beyond the diffraction limit along with an enhanced axial resolution for a fixed cell sample labeled with commercially available quantum dots. The inherent temporal averaging of the ISM technique enables image acquisition of the fluctuation correlation contrast within millisecond-scale pixel dwell times. SOFISM can therefore offer a robust path to achieve high-resolution images within a slightly modified confocal microscope, using standard fluorescent labels and reasonable acquisition times. (C) 2020 Optical Society of America under the terms of the OSA Open Access Publishing Agreement
Keywords:
STRUCTURED ILLUMINATION MICROSCOPY
FLUORESCENCE MICROSCOPY
LATERAL RESOLUTION
ENHANCED SOFI
RECONSTRUCTION
LIMIT
EXCITATION
ALGORITHM
BLINKING
PROTEIN
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