Reduction of coherent artefacts in super-resolution fluorescence localisation microscopy.
Reduction of coherent artefacts in super-resolution fluorescence localisation microscopy.
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DOI：
10.1111/jmi.12453
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发表时间：
2016-12
影响因子：
2
通讯作者：
Waigh TA
中科院分区：
文献类型：
--
作者：
Georgiades P;Allan VJ;Dickinson M;Waigh TA
超分辨率定位显微镜技术依赖于整个视场的均匀照明，否则分辨率会降低，导致成像伪影，例如条纹。由于激光功率高、带宽窄，目前是 PALM/STORM 方法中切换荧光团的首选光源。然而，这些光源的高相干性常常会在显微镜中产生干涉现象，并在图像中产生相关的条纹/散斑伪影。我们定量地证明了使用聚合物膜散斑扰频器来减少相干现象的影响。表征了照明平面中、相机处以及荧光团软件定位后的散斑效应。散斑现象会降低重建图像中大长度尺度下显微镜的分辨率，散斑扰乱器抑制了这种效应，但小长度尺度分辨率在~30 nm 时没有变化。
Super‐resolution localisation microscopy techniques depend on uniform illumination across the field of view, otherwise the resolution is degraded, resulting in imaging artefacts such as fringes. Lasers are currently the light source of choice for switching fluorophores in PALM/STORM methods due to their high power and narrow bandwidth. However, the high coherence of these sources often creates interference phenomena in the microscopes, with associated fringes/speckle artefacts in the images. We quantitatively demonstrate the use of a polymer membrane speckle scrambler to reduce the effect of the coherence phenomena. The effects of speckle in the illumination plane, at the camera and after software localisation of the fluorophores, were characterised. Speckle phenomena degrade the resolution of the microscope at large length scales in reconstructed images, effects that were suppressed by the speckle scrambler, but the small length scale resolution is unchanged at ∼30 nm.
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影响因子：
48
作者：
Dempsey, Graham T.;Vaughan, Joshua C.;Chen, Kok Hao;Bates, Mark;Zhuang, Xiaowei
通讯作者：
Zhuang, Xiaowei
影响因子：
3.4
作者：
Bosch, Peter J.;Correa, Ivan R., Jr.;Subramaniam, Vinod
通讯作者：
Subramaniam, Vinod
影响因子：
3.7
作者：
Dulin D;Barland S;Hachair X;Pedaci F
通讯作者：
Pedaci F
影响因子：
2.5
作者：
Guan, Y. Q.;Cai, Y. Y.;Opas, M.
通讯作者：
Opas, M.
影响因子：
48
作者：
Rust, Michael J.;Bates, Mark;Zhuang, Xiaowei
通讯作者：
Zhuang, Xiaowei