immunofluorescence microscopy
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Immunofluorescence Microscopy. Immunofluorescence Microscopy:. When an antibody, or the antiimmunoglobulin antibody used to detect the antibody is labeled with a fluorescent dye This method is used when looking at the subcellular location of a protein of interest.

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Presentation Transcript
immunofluorescence microscopy1
Immunofluorescence Microscopy:
  • When an antibody, or the antiimmunoglobulin antibody used to detect the antibody is labeled with a fluorescent dye
  • This method is used when looking at the subcellular location of a protein of interest
immunofluorescence microscopy2
Immunofluorescence Microscopy:
  • See Page 162 in textbook for another diagram
technique
Technique:
  • Common dyes: fluorescein, rhodamine
  • Dyes chosen are excited by a certain light wavelength, usually blue or green, and emit light of a different wavelength in the visible spectrum
    • Eg. Fluorescein emits green light
    • Eg. Rhodamine emits orange/red light
  • By using selective filters in a fluorescence microscope only the light from the dye is detected
  • Available fluorescent labels now include red, blue, cyan or yellow fluorescent proteins
slide5
Uses:
  • This can be used to detect the distribution of any protein
  • By attaching different dyes to different antibodies the distribution of two or more molecules can be determined in the same cell or tissue sample
technology
Technology:
  • Confocal fluorescent microscope: uses a computer-aided techniques to produce a extremely thin optical section of a cell or tissue
    • Gives high resolution microscopy without elaborate sample preparation
technology continued
Technology Continued:
  • Time-lapse video microscopy: sensitive digital video cameras record the movement of fluorescently labeled molecules in cell membranes and their redistribution when cells come into contact with one another
references
References:
  • Class textbook (Human Molecular Genetics 3)
  • www.antibodystation.com/immunofluorescence-microscopy/
  • National University of Singapore website www.med.nus.edu.sg/path/services/immunofluor.htm
  • BD Biosciences website www.bdbiosciences.com
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