Single molecule light field microscopy
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jats:pWe introduce single molecule light field microscopy (SMLFM), a new class of three-dimensional (3D) single molecule localization microscopy. By segmenting the back focal plane of a microscope objective with an array of microlenses to generate multiple 2D perspective views, the same single fluorophore can be imaged from different angles. These views, in combination with a bespoke fitting algorithm, enable the 3D positions of single fluorophores to be determined from parallax. SMLFM achieves up to 20 nm localization precision throughout an extended jats:inline-formula <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" display="inline"> mml:mn6</mml:mn> <mml:mspace width="thinmathspace" /> <mml:mspace width="thinmathspace" /> mml:mtextµ</mml:mtext> <mml:mrow class="MJX-TeXAtom-ORD"> <mml:mi mathvariant="normal">m</mml:mi> </mml:mrow> </mml:math> </jats:inline-formula> depth of field. The capabilities of SMLFM are showcased by imaging membranes of fixed eukaryotic cells and DNA nanostructures below the optical diffraction limit.</jats:p>
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2334-2536
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The Royal Society (uf120277)
Royal Society (URF\R\180029)
Royal Society (RGF\EA\181021)
Wellcome Trust (212936/Z/18/Z)
Engineering and Physical Sciences Research Council (EP/G037256/1)
Engineering and Physical Sciences Research Council (EP/L015455/1)
Engineering and Physical Sciences Research Council (EP/R025398/1)