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Presentación 6

Fluorescence microscopy utilizes the property of certain atoms and molecules to absorb and emit light, allowing for the observation of cellular processes and biomolecular interactions. Samples must be stained with fluorophores to visualize specific structures, and various types of fluorescence microscopy, including epifluorescence, confocal, and multiphoton microscopy, provide different imaging capabilities. These techniques are essential for studying biological structures in both live and fixed samples.

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7 views2 pages

Presentación 6

Fluorescence microscopy utilizes the property of certain atoms and molecules to absorb and emit light, allowing for the observation of cellular processes and biomolecular interactions. Samples must be stained with fluorophores to visualize specific structures, and various types of fluorescence microscopy, including epifluorescence, confocal, and multiphoton microscopy, provide different imaging capabilities. These techniques are essential for studying biological structures in both live and fixed samples.

Uploaded by

j.luisborras68
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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FLUORESCENCE MICROSCOPY:

Fluorescence is the property of certain atoms and molecules to


absorb radiation at a specific wavelength and subsequently
emit light at a longer wavelength after the atoms are excited.

Applications: Used to observe autofluorescence in tissues of


bacteria, animals, and plants, and currently enables the study
of specific cellular processes, localization of molecules within
cells, and analysis of biomolecular interactions.

Fundamental Principle:
- Samples of interest must be stained with fluorescent dyes
called fluorophores to be observed under a fluorescence
microscope, as they do not naturally emit fluorescence.
- High-energy light passes through filters to eliminate non-
specific wavelengths.
- The fluorophore absorbs short-wavelength radiation and
emits long-wavelength radiation, generating a colored
image on a dark background.
TYPES OF FLUORESCENCE MICROSCOPY:

Epifluorescence Microscopy: It allows the visualization of biological


structures labeled with fluorochromes, facilitating the observation of
specific cellular components.

Cell nuclei, actin cytoskeleton and cell membrane.

Confocal Microscopy: Useful for obtaining three-dimensional images


of internal structures of cells, tissues, or materials due to its ability to
eliminate blur.

Detail of fibroblast and cellular cytoskeleton.

Multiphoton Microscopy: Images reveal intricate details of biological


structures in live or fixed samples, enabling the observation of dynamic
processes in tissues and cells with exceptional three-dimensional
resolution.

Blood vessels in living tissue.

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