Dual-color simultaneous structured illumination microscopy based on galvo-mirrors

Yuan, Yifan; Liu, Wenjie; Wang, Yueying; Lü, Yang; Xu, Fan; Hao, Xiang; Han, Yubing; Kuang, Cuifang; Liu, Xü

doi:10.1016/j.optcom.2022.128012

Cited by 4 publications

(1 citation statement)

References 18 publications

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“…The other limitation of a DMD or indeed any grating-based SIM instrument is that the optical path defines the separation between the ± 1st order beams, which must match or be smaller than the back-aperture of the imaging objective lens. This defined beam separation means that if another objective lens with a different back-aperture size was needed to be used the optical beam paths would have to be changed In part to address this, approaches that use individual control of the two in the sample plane interfering beams have been demonstrated recently using galvanometric or piezoelectric scan mirrors and piezoelectric stages for control of the grating orientation and phase, respectively [24][25][26]. Either two sets of two-axis galvanometric scanners are employed to position the two interference beams, or a single scanning mirror and retroreflecting prism are utilized, requiring precise alignment of multiple elements to space the beams appropriately.…”

Section: Introductionmentioning

confidence: 99%

Miniaturised structured illumination microscopy using two 3-axis MEMS micromirrors

Tinning

Donnachie

Christopher

et al. 2022

Preprint

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We present the development and performance characterisation of a novel structured illumination microscope (SIM) in which the grating pattern is generated using two optical beams controlled via 2 micro-electro-mechanical system (MEMS) three-axis scanning micromirrors. The implementation of MEMS micromirrors to accurately and repeatably control angular, radial and phase positioning delivers flexible control of the fluorescence excitation illumination, with achromatic beam delivery through the same optical path, reduced spatial footprint and cost-efficient integration being further benefits. Our SIM architecture enables the direct implementation of multi-colour imaging in a compact and adaptable package. The two-dimensional SIM system approach is enabled by a pair of 2 mm aperture electrostatically actuated three-axis micromirrors having static angular tilt motion along the x- and y-axes and static piston motion along the z-axis. This allows precise angular, radial and phase positioning of two optical beams, generating a fully controllable spatial interference pattern at the focal plane by adjusting the positions of the beam in the back-aperture of a microscope objective. This MEMS-SIM system was applied to fluorescent bead samples and cell specimens, and was able to obtain a variable lateral resolution improvement between 1.3 and 1.8 times the diffraction limited resolution.

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Section: Introductionmentioning

confidence: 99%

Miniaturised structured illumination microscopy using two 3-axis MEMS micromirrors

Tinning

Donnachie

Christopher

et al. 2022

Preprint

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Ultra-high spatio-temporal resolution imaging with parallel acquisition-readout structured illumination microscopy (PAR-SIM)

Xu,

Wang,

Qiao

et al. 2024

Light Sci Appl

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Structured illumination microscopy (SIM) has emerged as a promising super-resolution fluorescence imaging technique, offering diverse configurations and computational strategies to mitigate phototoxicity during real-time imaging of biological specimens. Traditional efforts to enhance system frame rates have concentrated on processing algorithms, like rolling reconstruction or reduced frame reconstruction, or on investments in costly sCMOS cameras with accelerated row readout rates. In this article, we introduce an approach to elevate SIM frame rates and region of interest (ROI) coverage at the hardware level, without necessitating an upsurge in camera expenses or intricate algorithms. Here, parallel acquisition-readout SIM (PAR-SIM) achieves the highest imaging speed for fluorescence imaging at currently available detector sensitivity. By using the full frame-width of the detector through synchronizing the pattern generation and image exposure-readout process, we have achieved a fundamentally stupendous information spatial-temporal flux of 132.9 MPixels · s−1, 9.6-fold that of the latest techniques, with the lowest SNR of −2.11 dB and 100 nm resolution. PAR-SIM demonstrates its proficiency in successfully reconstructing diverse cellular organelles in dual excitations, even under conditions of low signal due to ultra-short exposure times. Notably, mitochondrial dynamic tubulation and ongoing membrane fusion processes have been captured in live COS-7 cell, recorded with PAR-SIM at an impressive 408 Hz. We posit that this novel parallel exposure-readout mode not only augments SIM pattern modulation for superior frame rates but also holds the potential to benefit other complex imaging systems with a strategic controlling approach.

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Using 3D printed optical elements for multifocal image scanning microscopy

Christopher,

Donnachie,

Rooney

et al. 2024

Multiscale Imaging and Spectroscopy V

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Dual-color simultaneous structured illumination microscopy based on galvo-mirrors

Cited by 4 publications

References 18 publications

Miniaturised structured illumination microscopy using two 3-axis MEMS micromirrors

Miniaturised structured illumination microscopy using two 3-axis MEMS micromirrors

Ultra-high spatio-temporal resolution imaging with parallel acquisition-readout structured illumination microscopy (PAR-SIM)

Using 3D printed optical elements for multifocal image scanning microscopy

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