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Confocal microscopy is included within optical microscopy, specifically of fluorescence miscroscopy. The development of new fluorescent probes and the increasing of resolution provided by the use of confocal microscopy, have made possible that fluorescence is one of the most used techniques nowadays. Confocal microscopy is characterised by:

  • The use of a temporary focused light source on a single point of the sample, one laser, for this reason the final image of the sample is obtained after successive scans of the sample.
  • The accessibility of the radiation emitted to the photodetector is limited by a diaphragm (pinhole), which allows the exclusion of the radiation emitted by the sample points that are not in the focal plane. Furthermore, the use of an appropriate software allows to improve the image quality by decreasing of the sensitivity of the photodetectors to low intensities of fluorescence, reducing the background noise.

The combination of capturing images in the focal plane with the software allows the use of the confocal microscope as an "optical microtome" which enables to obtain optical sections of the sample, basis for obtaining three-dimensional images.

 
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