Vortex light field microscopy: 3D spectral single-molecule imaging with a twist

被引:1
作者
Zhang, Boya [1 ]
Daly, Sam [2 ]
Zhu, Chengxi [1 ]
Lenz, Martin o. [1 ]
Weiss, Lucien e. [3 ]
Needham, Lisa-maria [1 ]
Peters, Ruby [4 ,5 ]
Lee, Steven f. [2 ]
O'holleran, Kevin [1 ]
机构
[1] Univ Cambridge, Cambridge Adv Imaging Ctr, Downing Site, Cambridge CB2 3DY, England
[2] Univ Cambridge, Yusuf Hamied Dept Chem, Lensfield Rd, Cambridge CB2 1EW, England
[3] Polytech Montreal, Dept Engn Phys, Montreal, PQ H3T 1J4, Canada
[4] Univ Cambridge, Dept Physiol Dev & Neurosci, Cambridge CB2 3EL, England
[5] Univ Sheffield, Sch Math & Phys Sci, Sheffield S3 7RH, England
基金
英国工程与自然科学研究理事会; 英国生物技术与生命科学研究理事会;
关键词
DIFFRACTION-LIMIT; SUPERRESOLUTION; SPECTROSCOPY; EXCITATION;
D O I
10.1364/OPTICA.534148
中图分类号
O43 [光学];
学科分类号
070207 ; 0803 ;
摘要
3D single-molecule imaging reveals nanoscale structures in cell volumes but is limited by the need for spectrally distinct fluorophores. We address this limitation with vortex light field microscopy (VLFM), a 3D spectral single-molecule localization technique with 25 nm spatial and 3 nm spectral precision over a 4 mu m depth of field. By modifying our previous single-molecule light field microscope with an azimuthally oriented prism array, we generated spectral disparity orthogonal to axial disparity, enabling simultaneous spatial and spectral localization on a single detector. We demonstrate VLFM with four-color 3D single-particle tracking and two-color 3D dSTORM imaging in fixed cells, successfully identifying dyes with spectral peaks just 15 nm apart. This shows VLFM's potential for enhancing spatial biology workflows requiring highly multiplexed imaging.
引用
收藏
页码:1519 / 1525
页数:7
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