Wicklund_111207 - Fluorescent proteins expressed...

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Fluorescence Bioimaging in Translational Research Kim Wicklund, PhD Research Imaging Product Manager Scientific Equipment Group
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Outline What is fluorescence? How can we put it in a biological system? How can we see it?
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Light Electromagnetic energy, photons
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Fluorescence ¾ Excitation by photon ¾ Relaxation ¾ Emission of a photon at longer wavelength (Stokes shift)
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Fluorescence Spectra
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Fluorescence in Biology Autofluorescence ¾ aromatic amino acids, NADPH ¾ in cells, arises from mitochondria and lysosomes ¾ in animals, keratin and collagen Fluorophore attached to antibodies for staining specific structure (dead cells/tissue)
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Unformatted text preview: Fluorescent proteins expressed genetically (live cells/tissues/animals) Fluorescent Proteins named after color of light that is emitted e.g., GFP Green Fluorescent Protein Use of Fluorescent Proteins (FP) Transfection - Insert FP genetic code into cellular DNA upstream of a protein of interest Protein of interest becomes labeled with FP Images of FP Labeled Organelles/Structures Optical Sectioning Fluorescence Microscope Lateral (XY) resolution: ~ 0.25 mm Axial (Z) resolution: > 1 mm Live Cell Imaging...
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This note was uploaded on 03/09/2008 for the course BIOS 95 taught by Professor Varies during the Spring '08 term at Lehigh University .

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Wicklund_111207 - Fluorescent proteins expressed...

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