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Results for “Fluorescence Microscopy”

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At least 109 records · Page 6Linked to original sources
PubMed2005

Characterization of sectioning fluorescence microscopy with thin uniform fluorescent layers: Sectioned Imaging Property or SIPcharts.

Thin, uniformly fluorescing reference layers can be used to characterize the imaging conditions in confocal, or more general, sectioning microscopy. Through-focus datasets of such layers obtained by standard microscope routines provide the basis for the approach. A set of parameters derived from these datasets is developed for defining a number of relevant sectioned imaging properties. The main characteristics of a particular imaging situation can then be summarized in a Sectioned Imaging Property-chart or SIPchart. We propose the use of such charts for the characterization of imaging properties in confocal and multiphoton microscopy. As such, they can be the basis for comparison of sectioned imaging condition characteristics, quality control, maintenance or reproduction of sectioned imaging conditions and other applications. Such charts could prove useful in documenting the more relevant properties of the instrumentation used in microscopy studies. The method carries the potential to provide the basis for a general characterization of sectioned imaging conditions as the layers employed can be characterized and fabricated to standard specifications. A limited number of such thin, uniformly fluorescing layers is available from our group for this purpose. Extension of the method to multiphoton microscopy is discussed.

Journal Article↗
PubMed2003

Visualizing membrane trafficking using total internal reflection fluorescence microscopy.

There is a dizzying array of fluorescent probes now commercially available to monitor cellular processes, and advances in molecular biology have highlighted the ease with which proteins can now be labelled with fluorophores without loss of functionality. This has led to an explosion in the popularity of fluorescence microscopy techniques. One such specialized technique, total internal reflection fluorescence microscopy (TIR-FM), is ideally suited to gaining insight into events occurring at, or close to, the plasma membrane of live cells with excellent optical resolution. In the last few years, the application of TIR-FM to membrane trafficking events in both non-excitable and excitable cells has been an area of notable expansion and fruition. This review gives a brief overview of that literature, with emphasis on the study of the regulation of exocytosis and endocytosis in excitable cells using TIR-FM. Finally, recent applications of TIR-FM to the study of cellular processes at the molecular level are discussed briefly, providing promise that the future of TIR-FM in cell biology will only get brighter.

Animals↗
PubMed1998

Near-field fluorescence microscopy of cells.

We demonstrate the surface sensitivity of near-field scanning optical microscopy by fluorescence imaging of membrane and bulk proteins in cells. We discuss instrument design considerations for successful cell-biology work with NSOM and show that the technique is most suited for studying membrane proteins.

Animals↗
PubMed2001

Applications of fluorescence microscopy to studies of dental hard tissue.

In this review we describe applications of fluorescence microscopy to investigations of dental hard tissue. Many studies have shown that human teeth show autofluorescence and several applications of fluorescence microscopy have been reported, although the chemical nature of autofluorescence remains unknown. However, most applications are based on a static measurement mode, which has inherent limitations. These limitations can be overcome by the use of time-resolved fluorescence microscopy. Therefore, we attempted to combine ordinary and time-resolved fluorescence microscopy to study human dentine. Using stationary measurements, blue fluorescence with a peak of 440 +/- 10 nm and a width of approximately 100 nm was observed. Increases in fluorescence intensity were found to be dependent upon age and temperature, regardless of tooth type or gender. The results suggest that the aging mechanism for crown dentine differs from that for root dentine and that the observed increase in fluorescence intensity is partially due to a temperature-dependent physicochemical mechanism. We speculate that fluorescence microscopy will become a useful tool for analysis of dental hard tissue in several fields.

Aging↗
PubMed2006

High-resolution intravital NADH fluorescence microscopy allows measurements of tissue bioenergetics in rat ileal mucosa.

OBJECTIVES: NADH fluorescence microscopy has been used as an index of the metabolic state of tissue but is associated with various obstacles such as low spatial resolution and quenching effects of blood pigments that prevent reliable monitoring of tissue bioenergetics. The objective of this study was to develop a system to monitor tissue bioenergetics in vivo using NADH fluorescence microscopy in the rat ileal mucosa. MATERIALS AND METHODS: Using an inverted microscope with an epifluorescence unit and an intensified charge-coupled device camera, NADH fluorescence images were visualized. Fluorescence intensity was measured of beta-NADH solutions at varying concentration (n = 6) and pH (n = 3) and in ex vivo (n = 6) and in vivo (n = 6) preparations of ileal mucosa of Sprague-Dawley rats anesthetized with isoflurane. RESULTS: Intravital fluorescence microscopy reveals a map of the microcirculation that permits visualization of NADH fluorescence and intercapillary areas. The system was adjusted so a linear relationship between physiological concentrations of beta-NADH and fluorescence was achieved (r(2) = 0.98, p < .0001). Decreasing the pH of the solution had no effect on fluorescence intensity and fluorescence intensity in an anoxic ex vivo ileal segment was similar to that of the in vivo ileum after ischemia. Ischemia also resulted in spatial heterogeneity that was abolished by the addition of a 550-nm LP filter. CONCLUSIONS: With this system, intravital NADH fluorescence microscopy provides the high resolution necessary to reliably monitor tissue bioenergetics in the rat ileal mucosa.

Animals↗
PubMed1997

Target cell-induced calcium signals in human natural killer leukemia cells as revealed by confocal fluorescence microscopy.

The target cell-induced calcium response in natural killer (NK) cells was examined using indo-1 (a ratiometric fluorochrome), a newly developed video-rate ultraviolet laser-scanning confocal microscope system, and a human NK leukemia cell line, YTN. Susceptible target cells (JY cells), but not unsusceptible target cells (K562 cells), increased the intracellular calcium concentration ([Ca]i) in almost all indo-1-loaded YTN cells with a lag time, although the kinetics of increase in [Ca]i differed in individual YTN cells. The increase in [Ca]i was completely blocked by 2 mM EGTA and partially (around 50%) blocked by 10 microM nicardipine, the extents of inhibition of calcium response correlating well with those of inhibition of cytotoxic activity. Thus, the calcium response in YTN cells observed in this study may constitute a critical step in the manifestation of cytotoxic activity.

Biological Transport↗
PubMed1978

Polarization and fluorescent microscopy--simple methods for demonstrating pulmonary surfactant lipids.

Birefringence and fluorochrome lipid staining with benzpyrene are demonstrated as simple morphological methods to reveal the presence or absence of surfactant lipids in human newborn lung tissue. Lack of lipid birefringence proves to be an associated finding in the lungs of premature infants with hyaline membrane disease, indicating the possible pathogenetical importance of surfactant deficiency.

Benzopyrenes↗