Confocal Microscopy

Instrument: Leica TCS SP8

In standard fluorescence microscopy, samples are imaged by exciting them with light of one wavelength and detecting the emission of light of a different wavelength. However, the light passes into the object being imaged as far as it will go, thereby potentially exciting much of the object, so you pick up emission signals from much of the object at once and this can restrict how clear the images are.

Confocal microscopy involves the focusing of light to a specific depth of the specimen, which greatly improves the resolution of the imaging. By varying the depth during the imaging, you can effectively build up a 3-dimensional reconstruction of the item being imaged.

Confocal microscopy is typically used in biological investigations of cellular processes. Imaging of parts of a cell is made possible by introducing fluorescent chemicals into the cell that localise in particular organelles, or which attach to specific proteins. Confocal microscopy is an essential technique for cell biology research.
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