1 / 19

Wavelength and Light Properties

575 nm. Wavelength and Light Properties. (l). High Energy. Low Energy. n = c / l E = h n. Unvisible UV Light. Visible Blue Light. Discovery of the Fluorescence: 1852. Sir George Gabriel Stokes (1819-1903). Fruorite. Fluorescence. - escence - means "to become" in Latin .

nara
Download Presentation

Wavelength and Light Properties

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. 575 nm Wavelength and Light Properties (l) High Energy Low Energy n = c / l E = hn

  2. Unvisible UV Light Visible Blue Light Discovery of the Fluorescence: 1852 Sir George Gabriel Stokes (1819-1903) Fruorite Fluorescence - escence - means "to become" in Latin. - Fluor - comes from the Fluorite (fluorspar)

  3. Fluorescence

  4. Spectral Shift & Filters ( Stokes Shift)

  5. Fluorochrome (khroma crwmacolor) a fluorescent molecule Fluorophore (-phore jorozto bear, carrying) a component of a molecule which causes a molecule to be fluorescent Fluorescent dye XXX is labeled with a fluorescent dye … Fluorescent probe XXX is the Fluorescent probe for ...

  6. Fluorochromes Fluorescein isothiocyanate (FITC) Hoechst

  7. Quantum Yield(efficiency) QY (QE) the ratio of the number of photons emitted to the number of photons absorbed. Molar extinction coefficient ME the ability of a molecule to absorb light Fluorochromes

  8. Fluorochromes Quantum Yield total integrated photon emission over the entire spectral band of the fluorophore S1>S2

  9. Fluorochromes FITC ME - 93 000 QY - 0,87 Phycoerythrin (PE) ME - 1 960 000 QY - 0,82 Drs E.D. Wieder and L. Gibson, Gladstone Inst. Of Virology & Immunology, San-Francisco, CA

  10. Hoechst Fluorescent probes for DNA

  11. Ab ’ Ab ’’ a b pA Immunolabeling Ag … Primary antibody (Ab ’ )-FITC Ag … AC ’ … Secondary Antibody ( Ab ’’ )-FITC Ag … AC ’ -Avidin (a) … Biotin (b)-FITC Ag … AC ’ … Protein A (pA)-FITC

  12. Fluorescent Proteins Ag … Antibody-Fluorochome Protein … Fluorochome (Fluorescent Fusion Protein) Green Fluorescent Protein

  13. Fluorescent probes for Metal Ions Fluo-3 Ca++

  14. Fluorochromes Phycoerythrin (phycoerithrobilin) Texas Red

  15. Texas Red Phycoerythrin Conjugated Fluorochromes Red 613

  16. FL1 FL2 FL3 Laser 488 nm Conjugated Fluorochromes

  17. Quantum dot

  18. Fluorochromes & Cytometer

  19. Cells (Beads etc) Fluorochromes Staining Instrumentation “the art and science of measurement” Data Analysis & Presentation Sorting Examples

More Related