We 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 $6\,\,\unicode{x00B5}{\rm m}$ depth of field. The capabilities of SMLFM are showcased by imaging membranes of fixed eukaryotic cells and DNA nanostructures below the optical diffraction limit.

中文翻译：

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单分子光场显微镜

我们介绍了单分子光场显微镜（SMLFM），这是一类新的三维（3D）单分子定位显微镜。通过用微透镜阵列分割显微镜物镜的后焦平面以生成多个2D透视图，可以从不同角度对同一单个荧光团成像。这些视图与定制拟合算法相结合，可以从视差中确定单个荧光团的3D位置。SMLFM在扩展的6，\，\ unicode {x00B5} {\ rm m} $景深范围内实现了高达20 nm的定位精度。SMLFM的功能通过固定真核细胞膜和低于光学衍射极限的DNA纳米结构的成像得以展示。