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Results for “Fluorescence Microscopy”

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At least 1,351 records · Page 75Linked to original sources
PubMed2006

Lateral organization of a membrane protein in a supported binary lipid domain: direct observation of the organization of bacterial light-harvesting complex 2 by total internal reflection fluorescence microscopy.

A unique method is described for directly observing the lateral organization of a membrane protein (bacterial light-harvesting complex LH2) in a supported lipid bilayer using total internal reflection fluorescence (TIRF) microscopy. The supported lipid bilayer consisted of anionic 1,2-dioleoyl-sn-glycero-3-[phospho-rac-(1'-glycerol)] (DOPG) and 1,2-distearoly-sn-3-[phospho-rac-(1'-glycerol)] (DSPG) and was formed through the rupture of a giant vesicle on a positively charged coverslip. TIRF microscopy revealed that the bilayer was composed of phase-separated domains. When a suspension of cationic phospholipid (1,2-dioleoyl-sn-glycero-3-ethylphosphocholine: EDOPC) vesicles (approximately 400 nm in diameter), containing LH2 complexes (EDOPC/LH2 = 1000/1), was put into contact with the supported lipid bilayer, the cationic vesicles immediately began to fuse and did so specifically with the fluid phase (DOPG-rich domain) of the supported bilayer. Fluorescence from the incorporated LH2 complexes gradually (over approximately 20 min) spread from the domain boundary into the gel domain (DSPG-rich domain). Similar diffusion into the domain-structured supported lipid membrane was observed when the fluorescent lipid (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-lissamine-rhodamine B sulfonyl: N-Rh-DOPE) was incorporated into the vesicles instead of LH2. These results indicate that vesicles containing LH2 and lipids preferentially fuse with the fluid domain, after which they laterally diffuse into the gel domain. This report describes for first time the lateral organization of a membrane protein, LH2, via vesicle fusion and subsequent lateral diffusion of the LH2 from the fluid to the gel domains in the supported lipid bilayer. The biological implications and applications of the present study are briefly discussed.

Bacterial Proteins↗
PubMed1987

Illumination and detection systems for quantitative fluorescence microscopy.

Systems are described for the illumination and detection of light microscope images of fluorescence in living cells at low levels of excitation. The illumination system is based on the use of an acousto-optic modulator to control the wavelength and power of the excitation beam. The detection system utilizes an image intensifier coupled with a television camera to acquire images at extremely low intensities. Both the illumination and detection systems have superior performance compared to existing devices.

Animals↗
PubMed2002

Vegetative incompatibility in the ascomycete Rosellinia necatrix studied by fluorescence microscopy.

We describe our examination of the cytological characteristics of the vegetative incompatibility reaction in a filamentous ascomycetes, Rosellinia necatrix, by analyzing the fluorescence emitted by ethidium bromide and acridine orange stained nuclei. Hyphal anastomosis between incompatible strains, which were field and single ascospore isolates, were observed with cell death showing fused hyphae, and nuclei debris which were intensified by staining with ethidium bromide. In contrast, the nuclei in a living cell were not intensified by staining with ethidium bromide but were intensified by staining with acridine orange. A strain was found which did not form a barrier reaction, but which could be shown to undergo cell death and therefore showed a positive vegetative incompatibility reaction. We also examined the vegetative incompatibility among five single ascospore isolates and the putative parent strain from the same perithecium; all strains were incompatible. These results strongly suggest that vegetative incompatibility in R. necatrix is regulated by many loci.

Acridine Orange↗
PubMed1999

Autofluorescence in human dentine in relation to age, tooth type and temperature measured by nanosecond time-resolved fluorescence microscopy.

Stationary and dynamic characteristics of autofluorescence in human dentine were examined with a nanosecond time-resolved fluorescence microphotometer. With ageing, the intensity of the fluorescence increased whereas the decay time decreased. There were no differences in fluorescence intensity, decay time or spectral profile among tooth types taken from the same individuals or from individuals of the same age. Fluorescence intensity was enhanced after the application of heat (37 and 50 degrees C), but remained unchanged when specimens were kept at 4 degrees C. These results suggest that the increase in fluorescence intensity was partially due to a temperature-dependent physicochemical mechanism.

Aged↗
PubMed1990

Confocal fluorescence microscopy: some applications in bone cell biology.

Three-dimensional information is necessary for the proper investigation of the interrelationships of bone cells, and of the complex interface between these cells and the bone matrix they form and destroy. The use of fluorescence confocal microscopy was explored in the determination of the distribution of immunolabelled actin and vinculin--cytoskeletal and attachment proteins--in isolated chick bone cells cultured on dentine, and in neonate rat and rabbit calvaria. Confocal imaging, compared with conventional fluorescence imaging, greatly enhanced the interpretation possible.

Actins↗
PubMed2005

Observation of the Yarrowia lipolytica peroxisome-vacuole dynamics by fluorescence microscopy with a single filter set.

We constructed the fusion of peroxisomal acyl-CoA oxidase 3 and the enhanced yellow fluorescent protein (EYFP) for fluorescent labeling of Yarrowia lipolytica peroxisomes. Using the spectral overlap between EYFP and FM4-64, we developed a procedure for simultaneous observation of Y. lipolytica peroxisomes and vacuoles with the single fluorescein isothiocyanate filter set. Using this procedure we were able to follow the Y. lipolytica peroxisome-vacuole dynamics under pexophagy conditions and show that Y. lipolytica peroxisomes are degraded in the vacuoles by a macropexophagic mechanism.

Acyl-CoA Oxidase↗
PubMed1976

[Acid mucopolysaccharides in human blood and bone marrow cells shown by fluorescence microscopy (author's transl)].

In lymphocytes, monocytes and basophil granulocytes of patients with mucopolysaccharidosis specific fluorescence of acid mucopolysaccharides exists after fixation of blood smears with 9-aminoacridin-hydrochloride and subsequent staining with pseudoisocyanine at pH 4.2. A total of eleven children with various types of mucopolysaccharidosis were investigated. Specificaly stained granula were demonstrated in 5 to 49 percent of all lymphocytes and monocytes. Acid mucopolysaccharides were for the first time also shown in red-cell precursors of the bone marrow of children with mucopolysaccharidosis.

Acridines↗
PubMed2005

Rotational motions of macro-molecules by single-molecule fluorescence microscopy.

Several complementary techniques have been developed to determine average orientation, dynamics on multiple time scales, and concerted rotational motions of individual fluorescent probes bound to biological macromolecules. In both protein domains and nucleic acids, tilting and wobble are relevant to their functional mechanisms. Here we briefly review methods to detect angles and rotational motions of single fluorophores and give an example of three-dimensional, total internal reflection, single-molecule fluorescence polarization applied to actin as it is translocated by conventional muscle myosin.

Actins↗