The Intermediate Optical System of Laser-Scanning Confocal Microscopes

Stelzer, Ernst H. K.

doi:10.1007/978-0-387-45524-2_9

Cited by 27 publications

(16 citation statements)

References 26 publications

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“…Methods of transmission electron microscopic cytochemistry, although giving rise to two-dimensional images, offer a resolving power that is about two orders of magnitude superior to light microscopy, including confocal laser scanning microscopy (CLSM; Stelzer 1995). Their application has made major contributions in obtaining basic information about the nuclear functional architecture.…”

Section: Introductionmentioning

confidence: 99%

The functional architecture of the nucleus as analysed by ultrastructural cytochemistry

Fakan

2004

Histochem Cell Biol

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Ultrastructural cytochemistry has been, for many years now, a major tool for investigating structure-function relationships in the cell nucleus. It has been essential in approaching the roles which different nuclear structural constituents can play in nuclear functions. This article briefly summarises transmission electron microscopic studies aimed at characterising in situ nuclear architectural domains and their involvement in main nuclear functions, such as DNA replication, hnRNA transcription and pre-mRNA processing. It discusses the importance of ultrastructural cytochemistry in high resolution analyses of intranuclear distribution of chromatin domains and their topological relationships with other structural interphase nuclear constituents. It puts forward the central role of the perichromatin region as a functional nuclear domain. Finally, it attempts to critically evaluate some future applications of ultrastructural investigations of the nucleus and stresses the importance of combining them with light microscopic analyses of living cells.

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Section: Introductionmentioning

confidence: 99%

The functional architecture of the nucleus as analysed by ultrastructural cytochemistry

Fakan

2004

Histochem Cell Biol

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“…The fundamental advantage of a confocal microscope is based on the capability to eliminate diffuse out-of-focus light by means of a pinhole. Taking optical slices along the z-axis (z-stacks) of a transparent specimen, the three-dimensional (3D) spatial structure of fluorescent objects can be imaged (Stelzer 2006). The diameter of the detection pinhole was set to one airy unit (diameter of the airy disc), which is a compromise between out-of-focus light elimination and signal sensitivity (Pawley 2006).…”

Section: Imaging Systemmentioning

confidence: 99%

Application of confocal laser scanning microscopy (CLSM) to visualize the effect of porous media wettability on unsaturated pore water configuration

et al. 2011

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Purpose Wettability affects water configuration and thereby transport processes and microbial activity in soil. Approaches to visualize the effect of porous media wettability on water films surrounding particles are rarely available in the literature. The aim of this study is therefore (1) to visualize the effect of wettability on area and connectivity of the water phase and (2) to develop a segmentation strategy to enable water films and bulk water to be differentiated. Materials and methods Wettability of silica sand was rendered by silanization using dichlorodimethylsilane. The resulting contact angle was measured using the sessile drop method. Furthermore, wettability was characterized by the water penetration time test in air-dry samples and at a volumetric water content of 8 vol.%. Sulforhodamine B was used to stain distilled water at a concentration of 40 mg/l. By means of the Wilhelmy plate method, the influence of the dye on the liquid surface tension was tested. Confocal laser scanning microscopy (CLSM) was used to visualize the area and connectivity of the fluorescent-dyed water phase and the thickness of water films in untreated and silanized samples at water contents of 8, 16, and 32 vol.%. Results and discussion The silanization significantly increased the contact angle of silica sand, whereas the surface tension of the dye solution did not differ significantly from that of undyed water. CLSM visualized the distribution of water, focused across the surface grain layer. Thresholding of fluorescence signal in two-dimensional intensity projections enables the discrimination between film and bulk water. It was shown that even subcritical water repellency (contact angle <90) leads to a decrease in area and connectivity of the water phase by affecting film instead of bulk water. CLSM detects decreasing effects of wettability with increasing water content, which were no longer significant at a water content of 32 vol.%. CLSM was found to be more sensitive for detecting the effects of wettability than the water drop penetration time test.Conclusions CLSM provides new insight into wettabilitydependent water configuration. One advantage over conventional microscopy arises from the capability to visualize water focused over the pore space. Compared to other three-dimensional imaging techniques, the advantage of CLSM is its simplicity. If only the optically accessible upper layer of the porous media is of interest, sample preparation, measurements, and image analysis can easily be carried out with a cost-and time-saving setup.

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“…The OBA was projected with a single large aspheric lens (CL, f = 39 mm, ∅= 60mm) onto the entry window of a large-diameter liquid light-guide (LLG, 5-mm ∅, NA0.59). Because optical fibers and light guides have high throughput in the visible spectral range, reflections at untreated end points become the largest sources of loss (Stelzer, 1995). A means of reducing these coupling losses is to index-match the optically non-treated ends of the liquid light guide to a custom glass window having anti-reflex coating on the outward-facing side only (see inset in Fig.…”

Section: Resultsmentioning

confidence: 99%

Efficient large core fiber-based detection for multi-channel two-photon fluorescence microscopy and spectral unmixing

Ducros

Hoff

Evrard

et al. 2011

Journal of Neuroscience Methods

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The Intermediate Optical System of Laser-Scanning Confocal Microscopes

Cited by 27 publications

References 26 publications

The functional architecture of the nucleus as analysed by ultrastructural cytochemistry

The functional architecture of the nucleus as analysed by ultrastructural cytochemistry

Application of confocal laser scanning microscopy (CLSM) to visualize the effect of porous media wettability on unsaturated pore water configuration

Efficient large core fiber-based detection for multi-channel two-photon fluorescence microscopy and spectral unmixing

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