Confocal fluorescence mode

The vesicles were observed on an inverted microscope in phase-contrast and confocal fluorescence modes, as follows. The white cloud of lipid was gently dispersed in the tube and introduced into an observation chamber. The chamber was filled with the same solution as the internal solution except that 0.1 M sucrose was replaced with 0.1 M glucose (external solution). Because of the difference in the reffactivity of the internal and external solutions, the contrast of the vesicle s images was enhanced in the phase-contrast mode. For the confocal fluorescence microscopy, a confocal scanner unit (CSU 10, Yokogawa, Japan) was used, and the lipophilic fluorescent dye Nile Red (Molecular Probes, Inc., OR, USA) was added to the starting lipid solution at 0.3 wt% of the lipid. 

Figure 14.3. A survey of read-out modes for confocal fluorescence detection technologies and their applications to biological systems. NA = nucleic acid.

A high-performance confocal fluorescence detection unit usable in either a single channel or multichannel mode. 

Figure 19. Correlated images of the same sample observed using cryogenic SEM (left) and scanning laser confocal light microscopy (right). The confocal image in fluorescent mode (right) shows a concentration of fluorescing components that correlates with the clay structure.

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]. 

Figure 1). Because Auorescence is observed as luminosity on a dark background, Auorescent constituents of the specimen can be seen even in extremely small amounts. There are several different modes of Auorescence microscopy, of which the most important is confocal fluorescence microscopy. 

As mentioned above, spectral imaging microscopy is a form of multidimensional fluorescent microscopy where a fluorescent emission spectrum is acquired at each coordinate location in the sample. This mode of imaging has been implemented for wide field, confocal, and two-photon laser scanning microscopy, and several excellent... 

Fig. 8.8 Optical images (at an active zone I) of isolated silver clusters electrodeposited onto ITO by means of the double-pulse method ( i = —1,550 mV 2 = —VOO mV, 25 s) [37] (a) Scanning confocal microscopy image (topography mode), (b) Raman/fluorescence image of the same sample area, (c) SEM image corresponding to (a) and (b)...

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