期刊
JOURNAL OF MICROSCOPY
卷 273, 期 1, 页码 3-25出版社
WILEY
DOI: 10.1111/jmi.12753
关键词
Fluorescence microscopy; HELM; illumination patterns; patterned excitation; photoswitchable molecules; RESOLFT; SPEM; superresolution
类别
资金
- National Science Foundation
- Sandler Family Supporting Foundation
- David and Lucile Packard Foundation
- National Institutes of Health [GM25101, GM31627]
- Deutsche Forschungsgemeinschaft [TRR166, TP B05]
- U.S. Department of Energy by University of California Lawrence Livermore National Laboratory [W-7405-Eng48]
Structured-illumination microscopy allows widefield fluorescence imaging with resolution beyond the classical diffraction limit. Its linear form extends resolution by a factor of two, and its nonlinear form by an in-principle infinite factor, the effective resolution in practice being determined by noise. In this paper, we analyse the noise properties and achievable resolution of linear and nonlinear 1D and 2D patterned SIM from a frequency-space perspective. We develop an analytical theory for a general case of linear or nonlinear fluorescent imaging, and verify the analytical calculations with numerical simulation for a special case where nonlinearity is produced by photoswitching of fluorescent labels. We compare the performance of two alternative implementations, using either two-dimensional (2D) illumination patterns or sequentially rotated one-dimensional (ID) patterns. We show that 1D patterns are advantageous in the linear case, and that in the nonlinear case 2D patterns provide a slight signal-to-noise advantage under idealised conditions, but perform worse than 1D patterns in the presence of nonswitchable fluorescent background.
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