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1.
Commun Biol ; 2: 74, 2019.
Article in English | MEDLINE | ID: mdl-30820469

ABSTRACT

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.


Subject(s)
Cryoelectron Microscopy/methods , Cytological Techniques/instrumentation , Cytological Techniques/methods , Microscopy, Fluorescence/methods , ATP-Binding Cassette Transporters/genetics , ATP-Binding Cassette Transporters/metabolism , Animals , CHO Cells , Cricetinae , Cricetulus , ErbB Receptors/chemistry , ErbB Receptors/genetics , ErbB Receptors/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Escherichia coli Proteins/genetics , Escherichia coli Proteins/metabolism , Fluorescent Dyes/chemistry , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Maleimides/chemistry , Reproducibility of Results
2.
Rev Sci Instrum ; 88(9): 093512, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28964232

ABSTRACT

A technique for the creation of free-standing cryogenic targets for laser-driven ion acceleration is presented, which allows us to create solid state targets consisting of initially gaseous materials. In particular, the use of deuterium and the methods for its preparation as a target material for laser-driven ion acceleration are discussed. Moving in the phase diagram through the liquid phase leads to the substance covering an aperture on a cooled copper frame where it is solidified through further cooling. An account of characterization techniques for target thickness is given, with a focus on deducing thickness values from distance values delivered by chromatic confocal sensors.

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