Principles and applications of laser confocal microscopy in biomedical research
Keywords:
Confocal laser scanning microscopy, fluorescence, confocality, Z-Stack, 3D imagingAbstract
The confocal microscopy is a widely used technique with a broad range of applications in biomedicine. It provides the possibility to acquire images from fixed tissue as well as from living cells with a high-space resolution. This relies on the basic principle of confocality, which allows to direct a beam of laser to a limited focal plane established by the user thus preventing the acquisition of out-of-focus information. This in turn improves significantly the spatial resolution of the acquired images (in particular in «z» or depth plane), generating more accurate information than that provided by conventional fluorescence microscopy. Confocal microscopy is particularly useful for acquiring and analyzing images from thick specimens, such as tissue samples containing structures like cells and for observing subcellular structures, which are either autofluorescent or have been targeted with fluorescent molecules. The main adventage of the this technique is that it generates high-resolution images from as less as 0.5 microns in depth and that together with the use of free-access image software, it allows to perform three-dimensional reconstructions. Continuous improvements to the technique have made of confocal microscopy a fairly amenable tool for scientists working in biomedical, biological and biotechnological areas. The purpose of this review is to provide a general view on the principles and applications of confocal microscopy in the field of biomedicine discussing its advantages as compared to those offered by conventional fluorescence microscopy. Confocal microscopy is a powerful tool in the field of biomedical research and is readily available in microscopy
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