Cyanobacteria on the stem of Antocerote sp (Anthocerophyta). Laser 405nm-473nm-559nm; Auto fluorescence.
Confocal Microscopy is a technology which permits observation with a greater resolution than can be achieved using conventional optical microscopes.
It uses a laser system that applies the light beam in a scanning manner on a small section of the specimen. The laser applied at a specific wavelength on the specimen causes the excited molecules to emit fluorescence at a greater wavelength than the one applied. The fluorescence in a specimen can be due to molecules which are naturally present (autofluorescence, as in the case of chlorophyll) or can be produced by molecules called fluorochromes, which are applied artificially to the specimen. There are a large number of specific fluorochromes on the market, used for different cellular structures and for different fluorescence emissions. The use of various combinations of lasers capable of detecting and producing fluorescence at different wavelengths permits scanning of a specimen within a broad range of the light spectrum, allowing for the observation of dyed structures with a degree of detail not possible with conventional techniques.
Because of its ability to easily penetrate the specimen, the confocal microscope captures images in different focal planes which, when linked to a computer program, reproduce a tridimensional image of the material observed.
The following characteristics have made confocal microscopy one of the preferred tools of scientists in the medical biological sciences and material sciences throughout the world:
High sensitivity in observations.
- Specificity in fluorescence emission
- Higher resolution
- Tridimensionality of images