Laser microscopy

Confocal microscopy can be realised either by object scanning ( on-axis ) or beam scanning ( off-axis ) a further distinction relates to single-beam and multiple-beam scanning systems. The small 

The advantages of the confocal scanning optical system over conventional microscopy are several fold (Table 5.18). Pawley [93] has described the fundamental limits of confocal microscopy. Confocal 

Confocal scanning microscopy can use non-laser and laser illumination sources, but in practice only the latter provide sufficient brightness. Confocal scanning optical microscopy (CSOM), in which the image is built up by synchronous scanning of the source and the detector units, can operate in transmission, reflection, or fluorescence mode. The lateral resolution of CSOM is of the order of 200 nm, which is a factor 1.4 better than that of the current optical techniques. CSOM images are usually sharper than those of conventional microscopy. The technique is some twenty years old [39]. 

Image processing by software enables generation of digital data sets which allow accurate quantitative measurements. Webb et al. [105] have dealt with 

The foundations of confocal scanned imaging in light microscopy have been reviewed [24]. Other reviews deal with confocal microscopy [106,107] and confocal laser microscopy [108]. CLSM has also been reviewed [109], in particular also for polymer science [110]. Several (hand)books on confocal microscopy [92,93,111] and on CSFM [92,111] are available. An early report on scanning laser microscopy has appeared [98] and history has been described [95]. 

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Ling X, Pritzker M D, Byerley J J and Burns C M 1998 Confocal scanning laser microscopy of polymer coatings J. Appl. Polym. Sc/. 67 149-58... 

Leonas K K 1998 Confocal scanning laser microscopy a method to evaluate textile structures Am. Dyest. Rep. 87 15-18 Wilson K R ef a/1998 New ways to observe and control dynamics Proc. SPIE 3273 214-18... 

X-ray difl raaion (structure grain size preferred orientation stress) Scanning laser microscopy Optical microscopy Oocnl thickness topography nucleation general morphology internal oxidation) l.R. spectroscopy (specialised analysis and applications)... 

TLC-Raman laser microscopy (X = 514 nm) in conjunction with other techniques (IR microscopy, XRF and HPLC-DAD-ESI-MS) has been used in the analysis of a yellow impurity in styrene attributed to reaction of the polymerisation inhibitor r-butylcatechol (TBC) and ammonia (from a washing step) [795]. Although TLC-FT-Raman did not allow full structural characterisation, several structural elements were identified. Exact mass measurement indicated a C20H25O3N compound which was further structurally characterised by 1H and 13C NMR. 

Verhaegh, N.A.M., and van Blaaderen, A. (1994) Dispersions of rhodamine-labeled silica spheres synthesis, characterization, and fluorescence confocal scanning laser microscopy. Langmuir 10, 1427-1438. 

Advances in pulsed lasers, microscopy and computer imaging, and the development of labelling techniques in which the donor and acceptor fluorophores become part of the biomolecules themselves have enabled the visualisation of dynamic protein interactions within living cells. 

M., and Nanninga, N. (1985) Three-dimensional chromatin distribution in neuroblastoma cell nuclei shown by confocal scanning laser microscopy. Nature 317,748-749. 

Focused X-ray beam (at Beam Line 15A) was used for this study. In order to improve the resolution, the focused beam (about 1 x 1.5 mm) was cut into 0.9-0.5 mm O by apertures which were set just in front of the cell. Modulation frequency was 10 Hz. Scanning X-Y stage which was originally developed for the laser microscopy was set perpendicular to the surface of the iron-base table and scanning and data acquisition were controlled by PC-9801 VM2 microcomputer (NEC Co. Ltd.) with the original program Various size and shape of metal foils were glued on the paper to have a model patterned sample. 

Barad, Y., Eisenberg, H., Horowitz, M., and Silberberg, Y. 1997. Nonlinear scanning laser microscopy by third harmonic generation. Appl. Phys. Lett. 70 922-24. 

Yomo, Urabe and coworkers (Yu et al, 2001), for example, reported the expression of a mutant GFP (actually the pET-21-GFPmutl-His6 mutant) in lecithin liposomes. Large GFP-expressing vesicles, prepared by the film hydration method, were analyzed using flow cytometry as well as confocal laser microscopy. 

The resolution of photoion laser microscopy is limited by two fundamental factors [7] the Heisenberg principle of uncertainty and the presence of the nonzero tangential component of the velocity of the ejected photoion (photoelectron). The same factors restrict the spatial resolution of the field-ion microscopy. It must be emphasized again that the key difference lies in the fact that for photoion microscopy there is no need for a strong (ionizing) electric field that distorts and desorbs the molecules. And also, the femtosecond laser radiation allows the photoion to be photoselectively extracted from certain parts of a molecule. 

The APMS used for this separation had an average particle size of 4-10 pm Normal phase HPLC of ferrocene and acetylferrocene performed with non-porous 1-3 pm spheres prepared in basic solution showed only one broad peak with no separation of the target molecules. Similarly, 20 pm spheres prepared in acidic solution showed no resolution of the ferrocenes (Figure 1). This indicates that particle size has some effect on the quality of the HPLC separation, but surface area is the major factor provided that the molecules to be separated can access the interiors of the mesoporous particles, which is dependent upon the pore size. (Experiments performed on APMS using confocal scanning laser microscopy indicated that these particles are porous throughout their interiors). 

Analyze by FACS or by pipeting the cells onto a microscope slide for confocal laser microscopy. 

Heertje, I., Vandervlist, P., Blonk, J.C.G., Hen-drickx, H., and Brakenhoff, G.J. 1987. Confocal scanning laser microscopy in food-research— Some observations. Food Microstructure 6 115-120. 

Heertje, 1., Nederlof, J., Hendrickx, H., and Lucas-senreynders, E.H. 1990. The observation of the displacement of emulsifiers by confocal scanning laser microscopy. Food Structure 9 305-316. 

LCM preferentially interact with tumor cells in vivo Data from confocal laser microscopy... 

Perhaps the most well-recognized fluorescent dye for detection of DNA hybridization is ethidium bromide (EtBr). EtBr is a cationic phenanthridinium compound that can bind to DNA by intercalation. This dye has an excitation maxima at 518 nm when bound to double-stranded DNA (dsDNA). Excitation of EtBr is often done by use of an argon ion laser, making this fluorophore a viable choice for applications in optical sensors as well as confocal scanning laser microscopy and fluorometry [41]. The structure of ethidium bromide is shown in Fig. 6. 

Figure 6 The co-localization of the carrier pCVI-HSA, identified with anti-HSA IgG (stained with FITC [A]) and HSC, identified with antidesmin and GFAP IgG (stained with Trite [B]) in fibrotic rat livers as assessed with confocal scanning laser microscopy. Note the co-localization as indicated by arrows. 

Auty, M.A.E., Fenelon, M.A., Guinee, T.P., Mullins, C., Mulvihill, D.M. 1999. Dynamic confocal scanning laser microscopy methods for studying milk protein gelation and cheese melting. Scanning 21, 299-304. 

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