Light microscopy confocal laser scanning

CD Circular dichroism Die Differential interference contrast DLS Dynamic light scattering FTIR Fourier-transform infrared LSCM Laser scanning confocal microscopy LCST Lower critical solution temperature Me Methyl... 

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.

Details are given of the production of a biodegradable lactide-caprolactone copolymer nanofibrous scaffold by electrospinning. Interactions between the scaffold and human coronary artery smooth muscle cells were demonstrated via MTS assay, phase contrast light microscopy, SEM, immunohistology assay and laser scanning confocal microscopy. 34 refs. 

In the present chapter, we discuss the principles and techniques commonly used for observing biological surface structures, including optical microscopy (light microscopy, laser scanning confocal microscopy), electron microscopy (scanning electron microscopy, transmission electron microscopy), and scanning probe microscopy. We describe and contrast the sample preparation of each technique. Quantitative data analysis as well as the limitations of each technique is also addressed. 

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-... 

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. 

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)... 

Two-photon excitation provides intrinsic 3-D resolution in laser scanning fluorescence microscopy. The 3-D sectioning effect is comparable to that of confocal microscopy, but it offers two advantages with respect to the latter because the illumination is concentrated in both time and space, there is no out-of-focus photo-bleaching, and the excitation beam is not attenuated by out-of-focus absorption, which results in increased penetration depth of the excitation light. 

---