SOFISM: Super-resolution optical fluctuation image scanning microscopy
Aleksandra Sroda, Adrian Makowski, Ron Tenne, Uri Rossman, Gur Lubin, Dan Oron, Radek Lapkiewicz
Abstract
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 <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" display="inline"> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mo>×</mml:mo> <mml:mn>2.5</mml:mn> </mml:mrow> </mml:math> 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.