1 / 9

Immunofluorescence Microscopy

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.

jessamine-hobbs
Download Presentation

Immunofluorescence Microscopy

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. ImmunofluorescenceMicroscopy

  2. 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

  3. Immunofluorescence Microscopy: • See Page 162 in textbook for another diagram

  4. 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

  5. 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

  6. 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

  7. The Zeiss LSM 410 confocal microscope

  8. 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

  9. 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

More Related