Increased localization precision by interference fringe analysis

Ebeling, Carl G.; Meiri, Amihai; Martineau, Jason; Zalevsky, Zeev; Gerton, Jordan M.; Menon, Rajesh

doi:10.1039/c5nr01927c

Cited by 9 publications

(4 citation statements)

References 31 publications

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“…Consequently, the current implementation of SDsSMLM has a reduced image resolution. This can be improved by replacing this grating with a new phase grating that can significantly suppress the 0th order and maximize the transmission efficiency only at the −1st and 1st orders, with the relatively high total transmission efficiency expected to be more than 85% 18 . In comparison, the blazed grating reported in our previous sSMLM system 5,6 has an absolute transmission efficiency of~18% for the 0th order and an absolute transmission efficiency of~50% for the 1st order in the far-red channel, corresponding to an overall efficiency of~68%.…”

Section: Discussionmentioning

confidence: 99%

Symmetrically dispersed spectroscopic single-molecule localization microscopy

et al. 2020

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Spectroscopic single-molecule localization microscopy (sSMLM) was used to achieve simultaneous imaging and spectral analysis of single molecules for the first time. Current sSMLM fundamentally suffers from a reduced photon budget because the photons from individual stochastic emissions are divided into spatial and spectral channels. Therefore, both spatial localization and spectral analysis only use a portion of the total photons, leading to reduced precisions in both channels. To improve the spatial and spectral precisions, we present symmetrically dispersed sSMLM, or SDsSMLM, to fully utilize all photons from individual stochastic emissions in both spatial and spectral channels. SDsSMLM achieved 10-nm spatial and 0.8-nm spectral precisions at a total photon budget of 1000. Compared with the existing sSMLM using a 1:3 splitting ratio between spatial and spectral channels, SDsSMLM improved the spatial and spectral precisions by 42% and 10%, respectively, under the same photon budget. We also demonstrated multicolour imaging of fixed cells and three-dimensional single-particle tracking using SDsSMLM. SDsSMLM enables more precise spectroscopic single-molecule analysis in broader cell biology and material science applications.

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Section: Discussionmentioning

confidence: 99%

Symmetrically dispersed spectroscopic single-molecule localization microscopy

et al. 2020

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“…However, it is well known that eventually the resolution limit is directly associated to the Signal To Noise ratio (SNR) 18 . This is obvious since the localization capability and precision is directly related to the SNR 19 . Thus, it could be advantageous to perform the temporal readouts coming from the probe at low temporal frequency, i.e.…”

Section: Temporal Super Resolution Via Time-spectral Polarization Encmentioning

confidence: 99%

Time-Spectral based Polarization-Encoding for Spatial-Temporal Super-Resolved NSOM Readout

Karelits

Mandelbaum

Zalevsky

et al. 2019

Sci Rep

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Detection of evanescent waves through Near-field Scanning Optical Microscopy (NSOM) has been simulated in the past, using Finite Elements Method (FEM) and 2D advanced simulations of a silicon Schottky diode, shaped as a truncated trapezoid photodetector, and sharing a subwavelength pin hole aperture. Towards enhanced resolution and next applications, the study of polarization’s influence was added to the scanning. The detector has been horizontally shifted across a vertically oriented Gaussian beam while several E-field modes, are projected on the top of the device. Both electrical and electro-optical simulations have been conducted. These results are promising towards the fabrication of a new generation of photodetector devices which can serve for Time-Spectral based Polarization-Encoding for Spatial-Temporal Super-Resolved NSOM Readout, as developed in the study.

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“…Localization microscopy (LM) [1][2][3][4][5][6][7][8][9][10][11] -sometimes called Single Molecule Localization Microscopy (SMLM) -has transformed biological fluorescence microscopy over the past decade and a half. Techniques like PALM [12][13][14][15] and STORM [16][17][18], together with STED [19] and structured illumination [20], have overcome the optical diffraction limit imposed by classical microscopes.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Spectral Differentiation of Multiple Fluorophores in Super-resolution Imaging Using a Glass Phase Plate

Fernando

Martineau

et al. 2022

Preprint

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Precision in superresolution microscopy is dependent on the photon yield. Multicolor localization microscopy typically relies on bandpass filters and sequential imaging to distinguish fluorescent tags. By engineering the pointspread function with a spatial light modulator (SLM), the wavelength of specific fluors can be distinguished by their unique pattern on the camera face and can thereby increase photon budgets and be imaged simultaneously. Here, instead of an SLM, we insert a silicon-dioxide phase plate, the X-phase plate (XPP) in the Fourier plane of the detection path of a wide-field fluorescence microscope to engineer the PSFs (XPSF). We demonstrate that the XPSF can be used for hyper-spectral superresolution microscopy of biological samples. The XPSF achieves ∼25 nm in-plane resolution, ∼250 nm axial resolution and can distinguish three fluorescent probes with ∼80 nm peak separation between consecutive spectra at the vicinity of axial focus. © 2022 Optical Society of America under the terms of the OSA Open Access Publishing AgreementOCIS codes(110.4234) Multispectral and hyperspectral imaging; (070.0070) Fourier optics and signal processing; (090.1970) Diffractive optics; (090.1970) Phase retrieval; (110.0180) Microscopy; (110.4235) Nanolithography; (180.2520) Fluorescence microscopy; (100.6640) Superresolution;http://dx.doi.org/10.1364/optica.XX.XXXXXX

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Increased localization precision by interference fringe analysis

Cited by 9 publications

References 31 publications

Symmetrically dispersed spectroscopic single-molecule localization microscopy

Symmetrically dispersed spectroscopic single-molecule localization microscopy

Time-Spectral based Polarization-Encoding for Spatial-Temporal Super-Resolved NSOM Readout

Simultaneous Spectral Differentiation of Multiple Fluorophores in Super-resolution Imaging Using a Glass Phase Plate

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