Imaging in scanning microscopes with slit‐shaped detectors

Wilson, Tony; Hewlett, S. J.

doi:10.1111/j.1365-2818.1990.tb03053.x

Cited by 37 publications

(20 citation statements)

References 8 publications

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“…The transverse resolution is defined as the transition width from the 10% to the 90% power points of the transverse response curve. These resolution measurements are consistent with calculations based on diffraction theory, as presented previously [9, 16, 25, 26] . In both cases, the background is higher (contrast is lower) for the line-scanned configuration due to the loss of confocality along one dimension.…”

supporting

confidence: 91%

“…In addition to improving the imaging speed, a line-scanned (LS) system can potentially increase the signal-to-noise ratio (SNR) compared to a point-scanned (PS) system by increasing pixel dwell times by two to three orders of magnitude for a given frame rate and field of view; however, photobleaching may ultimately limit the achievable SNR. There are also tradeoffs in imaging performance due to the loss of confocality along the focal line, resulting in a diminished optical sectioning capability [3, 4, 17, 25, 26] , and consequently a limited imaging depth.…”

mentioning

confidence: 99%

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Comparison of line-scanned and point-scanned dual-axis confocal microscope performance

et al. 2013

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The point-scanned dual-axis confocal (PS-DAC) microscope has been shown to exhibit a superior capability to reject out-of-focus and multiply scattered light in comparison to its conventional single-axis counterpart. However, the slow frame rate (typically < 5 Hz) resulting from point-by-point data collection makes these systems vulnerable to motion artifacts. While video-rate point-scanned confocal microscopy is possible, a line-scanned dual-axis confocal (LS-DAC) microscope provides a simpler means of achieving high-speed imaging through line-by-line data collection, but sacrifices contrast due to a loss of confocality along one dimension. Here we evaluate the performance tradeoffs between a LS-DAC and PS-DAC microscope with identical spatial resolutions. Characterization experiments of the LS-DAC and PS-DAC microscopes with tissue phantoms, in reflectance mode, are shown to match results from Monte-Carlo scattering simulations of the systems. Fluorescence images of mouse brain vasculature, obtained using resolution-matched LS-DAC and PS-DAC microscopes, demonstrate the comparable performance of LS-DAC and PS-DAC microscopy at shallow depths.

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confidence: 91%

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confidence: 99%

Comparison of line-scanned and point-scanned dual-axis confocal microscope performance

et al. 2013

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“…Patterns of varying types have been evaluated in the literature, both theoretically and experimentally (Wilson & Hewlett, 1990;Wilson, 1991;Conchello & Lichtman, 1994;Sandison & Webb, 1994;Jus Ïkaitis et al, 1996;Wilson et al, 1996;Fewer et al, 1997;Neil et al, 1997;Neil et al, 1998). The use of structured illumination patterns raises the further issues of how the structure of the pattern degrades with depth in a sample and what effect this has on the axial response.…”

Section: Discussionmentioning

confidence: 99%

An optical sectioning programmable array microscope implemented with a digital micromirror device

Hanley

Verveer

Gemkow

et al. 1999

Journal of Microscopy

113

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SummaryThe de®ning feature of a programmable array microscope (PAM) is the presence of a spatial light modulator in the image plane. A spatial light modulator used singly or as a matched pair for both illumination and detection can be used to generate an optical section. Under most conditions, the basic optical properties of an optically sectioning PAM are similar to those of rotating Nipkow discs. The method of pattern generation, however, is fundamentally different and allows arbitrary illumination patterns to be generated under programmable control, and sectioning strategies to be changed rapidly in response to speci®c experimental conditions. We report the features of a PAM incorporating a digital micromirror device, including the axial sectioning response to¯uorescent thin ®lms and the imaging of biological specimens. Three axial sectioning strategies were compared: line scans, dot lattice scans and pseudo-random sequence scans. The three strategies varied widely in light throughput, sectioning strength and robustness when used on real biological samples. The axial response to thin uorescent ®lms demonstrated a consistent decrease in the full width at half maximum (FWHM), accompanied by an increase in offset, as the unit cells de®ning the patterns grew smaller. Experimental axial response curves represent the sum of the response from a given point of illumination and cross-talk from neighbouring points. Cross-talk is minimized in the plane of best focus and when measured together with the single point response produces a decrease in FWHM. In patterns having constant throughput, there appears to be tradeoff between the FWHM and the size of the offset. The PAM was compared to a confocal laser scanning microscope using biological samples. The PAM demonstrated higher signal levels and dynamic range despite a shorter acquisition time. It also revealed more structures in x-z sections and less intensity drop-off with scanning depth.

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“…To enable us to make some comparisons among various systems, we therefore arbitrarily choose the half-width at half-maximum of the I(u) curves as a measure of the optical sectioning strength. In doing so, we should remember that we are considering a specific object and that not all objects possess identical sectioning strengths (Wilson and Hewlett 1990a).…”

Section: Introductionmentioning

confidence: 99%

Resolution enhancement in three-dimensional confocal microscopy

Hewlett

Wilson

1991

Machine Vis. Apps.

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Abstract:We discuss various methods of tuning and improving both the optical sectioning strength and the lateral resolution of the confocal scanning optical microscope. Techniques based on using different wavelengths, pupil plane filters, different sized detectors and laterally displaced detectors are considered and experimental results presented. Both bright field reflection and fluorescence imaging are discussed.

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Imaging in scanning microscopes with slit‐shaped detectors

Cited by 37 publications

References 8 publications

Comparison of line-scanned and point-scanned dual-axis confocal microscope performance

Comparison of line-scanned and point-scanned dual-axis confocal microscope performance

An optical sectioning programmable array microscope implemented with a digital micromirror device

Resolution enhancement in three-dimensional confocal microscopy

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