Direct‐view microscopy: optical sectioning strength for finite‐sized, multiple‐pinhole arrays

McCabe, Eithne M.; Fewer, D. T.; Ottewill, Adrian C.; Hewlett, S. J.; Hegarty, J.

doi:10.1111/j.1365-2818.1996.tb00004.x

Cited by 18 publications

(6 citation statements)

References 7 publications

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“…The combination of RESOLFT and structured illumination therefore promises <50-60 nm with sub-second image acquisition times of >50 × 50 μm 2 fields of view (Rego et al 2012). A more advanced illumination pattern than using parallel lines of high intensities as in conventional SIM is the scanning with thousands of parallelized points as in spinning-disc microscopy (McCabe et al 1996) or in multifocal SIM (York et al 2012). Similarly, more than 100 000 doughnuts were generated simultaneously for ultra-fast live-cell RESOLFT imaging of large fields of view with down to 70 nm spatial resolution (Chmyrov et al 2013) (Fig.…”

Section: Parallelizationmentioning

confidence: 99%

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Lens-based fluorescence nanoscopy

Eggeling

Willig

Sahl

et al. 2015

Quart. Rev. Biophys.

140

139

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The majority of studies of the living cell rely on capturing images using fluorescence microscopy. Unfortunately, for centuries, diffraction of light was limiting the spatial resolution in the optical microscope: structural and molecular details much finer than about half the wavelength of visible light (~200 nm) could not be visualized, imposing significant limitations on this otherwise so promising method. The surpassing of this resolution limit in far-field microscopy is currently one of the most momentous developments for studying the living cell, as the move from microscopy to super-resolution microscopy or 'nanoscopy' offers opportunities to study problems in biophysical and biomedical research at a new level of detail. This review describes the principles and modalities of present fluorescence nanoscopes, as well as their potential for biophysical and cellular experiments. All the existing nanoscopy variants separate neighboring features by transiently preparing their fluorescent molecules in states of different emission characteristics in order to make the features discernible. Usually these are fluorescent 'on' and 'off' states causing the adjacent molecules to emit sequentially in time. Each of the variants can in principle reach molecular spatial resolution and has its own advantages and disadvantages. Some require specific transitions and states that can be found only in certain fluorophore subfamilies, such as photoswitchable fluorophores, while other variants can be realized with standard fluorescent labels. Similar to conventional far-field microscopy, nanoscopy can be utilized for dynamical, multi-color and three-dimensional imaging of fixed and live cells, tissues or organisms. Lens-based fluorescence nanoscopy is poised for a high impact on future developments in the life sciences, with the potential to help solve long-standing quests in different areas of scientific research.

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Section: Parallelizationmentioning

confidence: 99%

“…A more advanced illumination pattern than using parallel lines of high intensities as in conventional SIM is the scanning with thousands of parallelized points as in spinning-disc microscopy (McCabe et al . 1996) or in multifocal SIM (York et al . 2012).…”

Section: The Coordinate-targeted Approachmentioning

confidence: 99%

Lens-based fluorescence nanoscopy

Eggeling

Willig

Sahl

et al. 2015

Quart. Rev. Biophys.

140

139

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“…To lower the excitation intensity and speed up the imaging acquisition time, it was proposed to use parallelized illumination instead of single-spot scanning. One approach called spinning disk confocal microscopy generates multifocal spots [137,138] by a Nipkow-type pinhole array disk and a microlens array [139], and the generated signal is projected onto a camera. Its pattern is similar to multifocal SIM shown in Figure 4E, but has an equal-pitch spiral shape to ensure the beam uniformity.…”

Section: Parallelized Illumination and Linescanning Confocal Microscopymentioning

confidence: 99%

Fluorescence imaging with tailored light

2019

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Fluorescence microscopy has long been a valuable tool for biological and medical imaging. Control of optical parameters such as the amplitude, phase, polarization, and propagation angle of light gives fluorescence imaging great capabilities ranging from super-resolution imaging to long-term real-time observation of living organisms. In this review, we discuss current fluorescence imaging techniques in terms of the use of tailored or structured light for the sample illumination and fluorescence detection, providing a clear overview of their working principles and capabilities.

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“…Alternatively, spinning disk systems [8][9][10][11][12] and programmable array systems [13][14][15][16] evaluate several lateral points in parallel. To reduce the cross talk between the lateral channels the pinholes have to be well separated, e.g., by ten times the pinhole diameter [17]. Array systems thus usually include a lateral scanning motion to cover the blank areas between the pinholes.…”

Section: Introductionmentioning

confidence: 99%

Chromatic confocal matrix sensor with actuated pinhole arrays

et al. 2015

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We present two versions of a chromatic confocal matrix sensor for the snapshot acquisition of three-dimensional objects. The first version contains separate illumination and detection pinhole arrays, while the second version uses a single pinhole array in double pass. The discrete lateral measurement points defined by the illumination and detection pinhole arrays are evaluated in parallel with a hyperspectral detection module. As this approach enables the spectrometric evaluation of all lateral channels, multilayer objects can be analyzed. To increase the lateral resolution the pinhole arrays are moved by micromechanical actuators. The paper includes a quantitative evaluation of the chromatic confocal module and proof-of-principle experiments with the full sensor system.

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Direct‐view microscopy: optical sectioning strength for finite‐sized, multiple‐pinhole arrays

Cited by 18 publications

References 7 publications

Lens-based fluorescence nanoscopy

Lens-based fluorescence nanoscopy

Fluorescence imaging with tailored light

Chromatic confocal matrix sensor with actuated pinhole arrays

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