Fast optical sectioning obtained by structured illumination microscopy using a digital mirror device

Xu, Dongli; Jiang, Tao; Li, Anan; Hu, Bihe; Feng, Zuren; Gong, Hui; Zeng, Shaoqun; Luo, Qi

doi:10.1117/1.jbo.18.6.060503

Cited by 42 publications

(39 citation statements)

References 12 publications

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“…A recent report demonstrated video-rate SIM for optical-sectioning imaging of large tissue areas, in which the maximum full-resolution frame-rate demonstrated was 16.6 Hz [15]. In this manuscript we improve upon those results, and demonstrate the feasibility of SIM for video-rate full-resolution (26.9 Hz at 2048 x 2048 pixel resolution) imaging of fluorescentlystained tissue surfaces.…”

Section: Discussionsupporting

confidence: 51%

Video-rate structured illumination microscopy for high-throughput imaging of large tissue areas

Schlichenmeyer¹,

Wang²,

Elfer³

et al. 2014

Biomed. Opt. Express

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Abstract:We report the development of a structured illumination microscopy instrument specifically designed for the requirements for higharea-throughput, optically-sectioned imaging of large, fluorescently-stained tissue specimens. The system achieves optical sectioning frame-rates of up to 33 Hz (and pixel sampling rates of up to 138.4 MHz), by combining a fast, ferroelectric spatial light modulator for pattern generation with the latest large-format, high frame-rate scientific CMOS camera technology. Using a 10X 0.45 NA objective and a 7 mm/sec scan stage, we demonstrate 4.4 cm 2 /min area-throughput rates in bright tissue-simulating phantoms, and 2 cm 2 /min area-throughput rates in thick, highly-absorbing, fluorescentlystained muscle tissue, with 1.3 μm lateral resolution. We demonstrate highcontrast, high-resolution imaging of a fluorescently-stained 30.4 cm 2 bovine muscle specimen in 15 minutes comprising 7.55 gigapixels, demonstrating the feasibility of the approach for gigapixel imaging of large tissues in short timeframes, such as would be needed for intraoperative imaging of tumor resection specimens.

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Section: Discussionsupporting

confidence: 51%

Video-rate structured illumination microscopy for high-throughput imaging of large tissue areas

Schlichenmeyer¹,

Wang²,

Elfer³

et al. 2014

Biomed. Opt. Express

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show abstract

“…Especially when high numerical aperture objective was used to obtain high resolution and the field of view was reduced to less than 500 × 500 μm 2 [7]. It was indispensable to image numbers of mosaics in order to cover the entire specimen area [8][9][10][11]. Repetitive imaging is cost-ineffective since the stage will be controlled to go through an acceleration-constant speed-deceleration process, which is inefficient.…”

Section: Introductionmentioning

confidence: 99%

Rapid imaging of large tissues using high-resolution stage-scanning microscopy

Yang

Zheng

Shang

et al. 2015

Biomed. Opt. Express

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Rapid and high-resolution imaging of large tissues is essential in biological research, like brain neuron connectivity research and cancer margins imaging. Here a novel stage-scanning confocal microscopy was developed for rapid imaging of large tissues. Line scanning methods and strip imaging strategy were used to increase the imaging speed. The scientific CMOS was used as line detector in sub-array mode and the optical sectioning ability can be easily adjusted by changing the number of line detectors according to different samples. Fluorescent beads imaging showed resolutions of 0.47 μm, 0.56 μm, and 1.56 μm in the X, Y, and Z directions, respectively, with a 40 × objective lens. A 10 × 10 mm 2 coronal plane with enough signal intensity could be imaged in about 88 sec at a sampling resolution of 0.16 μm/pixel. Rapid imaging of mouse brain slices demonstrated the applicability of this system in visualizing neuronal details at high frame rate.

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“…DMDs permit highly flexible codification of binary masks at frame rates above 20 kHz. Extensive application of the DMD to microscopy has been reported in the past few years including 60 conventional SIM microscopy with fringe projection [30], super-resolution and optical sectioning microscopy [31,32] and the programmable array microscope [33,34,35]. Interestingly, fast DMD and pattern illumination is at the core of optogenetics, a tool for noninvasive activation and silencing of neurons and muscles [36,37].…”

mentioning

confidence: 99%

Dual-mode optical microscope based on single-pixel imaging

Rodríguez

Clemente

Tajahuerce

et al. 2016

Optics and Lasers in Engineering

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We demonstrate an inverted microscope that can image specimens in both reflection and transmission modes simultaneously with a single light source. The microscope utilizes a digital micromirror device (DMD) for patterned illumination altogether with two single-pixel photosensors for efficient light detection. The system, a scan-less device with no moving parts, works by sequential projection of a set of binary intensity patterns onto the sample that are codified onto a modified commercial DMD. Data to be displayed are geometrically transformed before written into a memory cell to cancel optical artifacts coming from the diamond-like shaped structure of the micromirror array. The 24-bit color depth of the display is fully exploited to increase the frame rate by a factor of 24, which makes the technique practicable for real samples. Our commercial DMDbased LED-illumination is cost effective and can be easily coupled as an add-on module for already existing inverted microscopes. The reflection and transmission information provided by our dual microscope complement each other and can be useful for imaging non-uniform samples and to prevent self-shadowing effects.

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Fast optical sectioning obtained by structured illumination microscopy using a digital mirror device

Cited by 42 publications

References 12 publications

Video-rate structured illumination microscopy for high-throughput imaging of large tissue areas

Video-rate structured illumination microscopy for high-throughput imaging of large tissue areas

Rapid imaging of large tissues using high-resolution stage-scanning microscopy

Dual-mode optical microscope based on single-pixel imaging

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