Confocal Laser Scanning Microscopy CLSM

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. 

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... 

Confocal laser scanning microscopy (CLSM) in conjunction with specific staining techniques is best suited to elucidate intracellular trafficking and localization. CLSM is a specific epifluorescence microscopical technique capable of optical cross-sectioning with a spatial resolution of 1 /urn and below [41, 42],... 

HAPEX, a bone analogue material, with similar properties to cortical bone, was added to a HDPE matrix in different volumes (20% and 40%) to produce composite materials [271]. Confocal laser scanning microscopy (CLSM) was then used to examine cell morphology on HAPEX and the surface characteristics produced by different machining protocols. 

Durrenberger, M.B., Handschin, S., Conde-Petit, B., and Escher, F. (2001). Visualization of food structure by confocal laser scanning microscopy (CLSM). Lebensmitt. Wissen. Technol. 34, 11-17. 

Lamprecht, A. Schafer, U.F. Lehr, C.M. Visualization and quantification of polymer distribution in microcapsules by confocal laser scanning microscopy (CLSM). Int. J. Pharm. 2000, 196 (2), 223-226. 

The multilayer coating of particles and formation of ultrathin microcapsules were verified by confocal laser scanning microscopy (CLSM, Leica) and atomic force microscopy (AFM, NanoScope). For AFM measurements, a drop of each sample was deposited onto the silicon support (with a PEl/PSS sublayer) and dried. For CLSM analysis, the coated particles and multilayer capsule suspensions were preliminary colored with rhodamin C. 

This review covers the formation, composition, structure, function and properties of the acquired pellicle. Specifically, the formation of pellicle is considered in terms of thermodynamic and kinetic aspects. The composition of the pellicle is reviewed in terms of the proteins, carbohydrates and lipids that have been identified using a range of analytical techniques. The ultrastructure of the pellicle is described in some detail from studies involving enamel slabs carried in the mouth, in which the subsequent pellicle was analysed by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM). The function of the pellicle is outlined in terms of its lubrication properties, its ability to act as a semi-permeable membrane and its overall protection of the underlying enamel surfaces. Since pellicle is formed at the interface between the enamel surface and the oral environment, the important process of bacterial attachment to the pellicle surface is described and the specific bacterial binding sites found in the pellicle are summarised. The influence of diet and nutrition on the pellicle layer is considered. The formation of extrinsic stain is discussed in particular, the role that chlorhexidine... 

Assess capsules for bacterial attachment by Confocal Laser Scanning Microscopy (CLSM). 

Confocal laser scanning microscopy (CLSM) is a very useful technique for the identification of suspensions. It uses a variable pinhole aperture or variable-width slit to illuminate only the focal plane by the apex of a cone of laser hght. Out-of-focus items are dark and do not distract from the contrast of the image. 

As with flow cytometry, multiparameter apoptosis assays may also be performed by confocal laser scanning microscopy (CLSM). Using the approach similar to that detailed above for flow cytometry, we have examined NADPH content, mitochondrial membrane potential (CMX Rosamine fluorescence), and mitochondrial mass (Mitotracker Green), by CLSM. Figure 3 shows an example of a typical multiparameter assay performed by confocal microscopy. 

Figure 4. Subcellular localization of the photosensitizer BPD-MA. OVCAR-5 cells were incubated in 92 nM BPD-MA for 3 h and 10 nM rhodamine 123, a mitochondrial probe, for 20 min. Imaging was performed using confocal laser scanning microscopy (CLSM). (A) Exclusively mitochondrial green fluorescence of rhodamine 123 (B) red BPD-MA fluorescence (C) overlay of A -i- B, where yellow indicates co-localization (D) DIC transmission image. The colocalization in (C) indicates that BPD-MA localizes to mitochondria, but also stains other subcellular structures.

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