CONFOCAL LASER

Kapitza H G 1996 Confocal laser scanning microscopy for optical measurement of the microstructure of surfaces and layers Tech. Mess. 63 136-41... 

Schroth D 1997 The confocal laser scanning microscopy. A new tool in materials testing Matehalpruefung 39 264 Chestnut M H 1997 Confocal microscopy of colloids Curr. Opin. Colloid Interface Sc/. 2 158-61... 

The classical polarizing light microscope as developed 150 years ago is still the most versatile, least expensive analytical instrument in the hands of an experienced microscopist. Its limitations in terms of resolving power, depth of field, and contrast have been reduced in the last decade, in which we have witnessed a revolution in its evolution. Video microscopy has increased contrast electronically, and thereby revealed structures never before seen. With computer enhancement, unheard of resolutions are possible. There are daily developments in the X-ray, holographic, acoustic, confocal laser scanning, and scanning tunneling micro-... 

FIG. 7 Confocal laser scanning microscopy image of a fonr-layer polyelectrolyte/CdTe(S) nanocrystal shell assembled on 1.5-p,m-diameter ME particles. The polyelectrolyte film consists of two bUayers of PAH and PSS. (From Ref. 76.)... 

FIG. 9 Confocal laser scanning micrograph of a hollow polymer capsule. The polymer capsule was obtained from polymer multilayer-templated FDA microcrystals after removal of the colloidal core. The FDA microcrystals were coated with SDS and 11 polyelectrolyte layers [(PAH/PSS)3/PAH/ (PSS/PAH-FITC)2]. (PAH-FITC = PAH labeled with fluorescein isothiocyanate.) The microcrystal core was removed by exposure of the coated microcrystals to ethanol, causing solubilization of FDA. 

Spatially resolved measurements, based on the confocal laser microscope and related techniques, have recently enabled direct detection of individual molecules, single nanoparticles, and molecular assemblies, leading to elucidation of the heterogeneous nature of these systems and its dependence on the individual environments. 

J. Bloem, M. Veningra, and J. Shepherd, Fully automatic determination of soil bacterium numbers, cell volumes, and frequencies of dividing cells by confocal laser-scanning microscopy and image-analysis, Appl. Environ. Microbiol. 61 926... 

Takami S., Getchell M. and Getchell T. (1995). Resolution of sensory and mucoid glycoconjugates with terminal O-galactose residues in the mucomicrovillar complex of the vomeronasal sensory epithelium by dual confocal laser-scanning microscopy. Cell Tiss Res 280, 211-216. 

CLSM Confocal laser scanning microscopy (see LSCM)... 

Spengler and Hubert (2002) describe a confocal laser scanning microscope used in conjunction with a TOF mass spectrometer, and also possessing ion imaging... 

Figure 4. DDC (A), serotonin (B), and tyrosine hydroxylase (C) immunore-activity in the posterior region of a wild-type Drosophila ventral ganglion. Tyrosine hydroxylase (TH) encodes the rate-limiting step in dopamine biosynthesis and is a marker for dopamine cells. B and C are the same CNS assayed for both serotonin and TH. M, medial dopamine neurons VL, ventrolateral serotonin neurons DL, dorsolateral dopamine neurons. Short unmarked arrows in C show vacuolated cells that do not contain DDC immunoreactivity. The immunoreactivity in these cells may represent a nonspecific cross-reactivity of the rat TH antibody. The length bar in A is 50 pM. The images are confocal projections generated on a Molecular Dynamics-2000 confocal laser scanning microscope.

Lacerda, L. et al. (2007) Intracellular trafficking of carbon nanotubes by confocal laser scanning microscopy. Advanced Materials, 19 (11),... 

Homeffer V, Forsmann A, Strupat K et al (2001) Localization of analyte molecules in MALDI preparations by confocal laser scanning microscopy. Anal Chem 73 1016-1022... 

Azuma, H., Otagiri, M., Effects of absorption enhancers on the transport of model compounds in Caco-2 cell monolayers assessment by confocal laser scanning microscopy, J. Pharm. Sci. 1997, 86, 779-785. 

Historically, this has been the most constrained parameter, particularly for confocal laser scanning microscopes that require spatially coherent sources and so have been typically limited to a few discrete excitation wavelengths, traditionally obtained from gas lasers. Convenient tunable continuous wave (c.w.) excitation for wide-held microscopy was widely available from filtered lamp sources but, for time domain FLIM, the only ultrafast light sources covering the visible spectrum were c.w. mode-locked dye lasers before the advent of ultrafast Ti Sapphire lasers. 

McConnell, G. (2004). Confocal laser scanning fluorescence microscopy with a visible continuum source. Opt. Express 12, 2844—50. 

Baschong W, Suetterlin R, Laeng RH. Control of autofluorescence of archival formaldehyde-fixed, paraffin-embedded tissue in confocal laser scanning microscopy (CLSM)./. Histochem. Cytochem. 2001 49 1565-1571. 

Fig. 13 Uptake of OVA-encapsulating y-PGA-Phe nanoparticles by DCs. DCs were incubated with Texas Red-labeled OVA (TR-OVA) alone (a) or TR-OVA encapsulated within fluorescein-labeled nanoparticles (TR-OVA/FITC-NPs) (b). The intracellular localization of OVA (red) and NPs (green) was observed by confocal laser scanning microscopy...

Akagi et al. demonstrated the use of nanoparticles composed of amphiphilic poly (amino acid) derivatives as vaccine delivery and adjuvants [62, 102-104]. To evaluate the uptake of OVA encapsulated within y-PGA-Phe nanoparticles (OVA-NPs) by DCs, murine bone marrow-derived DCs were incubated with 250 nm-sized OVA-NPs for 30 min at 37 °C. The cells were then analyzed by flow cytometry (FCM) and confocal laser scanning microscopy (CLSM). OVA-NPs were efficiently taken up into DCs, whereas the uptake of OVA alone was barely detectable at the same concentration of OVA (Fig. 13). OVA-NPs were more efficiently taken up than OVA alone by DCs, and the uptake of OVA-NPs was inhibited at 4 °C. These results suggest that OVA-NPs were phagocytosed mainly via endocytosis by the DCs. In the case of OVA alone, an approximately 30-fold... 

Fig. 9 Surface modification of cells with ssDNA-PEG-lipid. (a) Real-time monitoring of PEG-lipid incorporation into a supported lipid membrane by SPR. (r) A suspension of small unilamellar vesicles (SUV) of egg yolk lecithin (70 pg/mL) was applied to a CH3-SAM surface. A PEG-lipid solution (100 pg/mL) was then applied, (ii) Three types of PEG-lipids were compared PEG-DMPE (C14), PEG-DPPE (C16), and PEG-DSPE (C18) with acyl chains of 14, 16, and 18 carbons, respectively, (b) Confocal laser scanning microscopic image of an CCRF-CEM cell displays immobilized FITC-oligo(dA)2o hybridized to membrane-incorporated oligo(dT)20-PEG-lipid. (c) SPR sensorigrams of interaction between oligo(dA)2o-urokinase and the oligo (dT)2o-PEG-lipid incorporated into the cell surface, (i) BSA solution was applied to block nonspecific sites on the oligo(dT)20-incorporated substrate, (ii) Oligo(dA)20-urokinase (solid line) or oligo(dT)20-urokinase (dotted line) was applied...

Fig. 10 Confocal laser scanning microscope images of islets with urokinase (UK) immobilized on the membrane. The green fluorescence indicates positive immunostaining for UK. (a) Islets were modified with oligo(dT)2o-PEG-lipid (C16) or (b) oligo(dT)2o-PEG-lipid (C18) then, oligo (dA)2o-UK was added to the media, (c) Unmodified islets with (left) and without (right) oligo (dT)20-PEG-lipids added to the solution. Insets. Bright field images. Scale bars 100 pm...

Scanning electron microscopy, light microscopy, and confocal laser scanning microscopy together with FISH Randomly amplified polymorphic DNA (RAPD), Enterobacterial repetitive intergenic consensus sequence (ERIC-PCR)... 

Sole A, Gaju N, Esteve I (2003) The biomass dynamics of cyanobacteria in an annual cycle determined by confocal laser scanning microscopy. Scanning 25 1-7... 

Structural variations may be also produced at the microscopical scale and are able to produce significant improvements in our understanding of stressor effects. Observation of the biofilm architecture and characterisation of the different fractions (i.e. algae, bacteria, mucopolysaccharides) may be useful to identify particular effects of toxicants to selective components of the biofilm. The use of confocal laser scanning microscopy remains promising [25]. 

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