Solid immersion microscopy images cells under cryogenic conditions with 12 nm resolution(35 visite) Wang L, Bateman B, Zanetti-domingues L, Moores A, L , Astbury S, Spindloe C, Darrow M, C , Romano M, Needham S, R , Beis K, Rolfe D, J , Clarke D, T , Martin-fernandez M, L
Parole chiave: Super-Resolution Fluorescence Microscopy
Central Laser Facility, Research Complex at Harwell, Science and Technology Facilities Council, Rutherford Appleton Laboratory, Harwell, Didcot, Oxford, OX11 0QX, UK. Diamond Light Source, Harwell Science and Innovation Campus, Didcot, OX11 0DE, UK. Department of Life Sciences, Imperial College London, London, SW7 2AZ, UK.
Super-resolution fluorescence microscopy plays a crucial role in our understanding of cell structure and function by reporting cellular ultrastructure with 20-30 nm resolution. However, this resolution is insufficient to image macro-molecular machinery at work. A path to improve resolution is to image under cryogenic conditions. This substantially increases the brightness of most fluorophores and preserves native ultrastructure much better than chemical fixation. Cryogenic conditions are, however, underutilised because of the lack of compatible high numerical aperture objectives. Here, using a low-cost super-hemispherical solid immersion lens (superSIL) and a basic set-up we achieve 12 nm resolution under cryogenic conditions, to our knowledge the best yet attained in cells using simple set-ups and/or commercial systems. By also allowing multicolour imaging, and by paving the way to total-internal-reflection fluorescence imaging of mammalian cells under cryogenic conditions, superSIL microscopy opens a straightforward route to achieve unmatched resolution on bacterial and mammalian cell samples.