Signal, noise and resolution in linear and nonlinear structured-illumination microscopy

Journal of Microscopy
E A IngermanMats G L Gustafsson

Abstract

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. LAY DESCRIPTION: Structured-illumination microscopy (SIM) is a high-resolution light micro...Continue Reading

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Citations

Apr 3, 2021·Light, Science & Applications·Gang WenHui Li
Apr 27, 2021·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Rainer Heintzmann
Apr 27, 2021·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Hai GongMark A A Neil
Apr 27, 2021·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Miguel A BolandMark A A Neil
Aug 27, 2021·Current Protocols·Jessica Valli, Jeremy Sanderson

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