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NIH 3T3 fibroblasts stained for Filamentous-actin Phalloidin Alexa Fluor 488 nm

NIH 3T3 fibroblasts stained for Filamentous-actin Phalloidin Alexa Fluor 488 nm. A. G. Sostarecz, P.A. Janmey, data not published. Phalloidin Alexa Fluor 488 nm. blue. green. Courtesy of Invitrogen website. A. G. Sostarecz, P.A. Janmey, data not published. E = h c λ

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NIH 3T3 fibroblasts stained for Filamentous-actin Phalloidin Alexa Fluor 488 nm

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  1. NIH 3T3 fibroblasts stained for Filamentous-actin Phalloidin Alexa Fluor 488 nm A. G. Sostarecz, P.A. Janmey, data not published

  2. Phalloidin Alexa Fluor 488 nm blue green Courtesy of Invitrogen website A. G. Sostarecz, P.A. Janmey, data not published E = hc λ Longer wavelength; lower energy Alexa Fluor - Fluorescein

  3. http://probes.invitrogen.com/resources/education/tutorials/1Introduction/player.htmlhttp://probes.invitrogen.com/resources/education/tutorials/1Introduction/player.html

  4. - - - 210 µm x 170 µm NIH 3T3 fibroblasts labeled with fluorescent TR- PIP2 A. G. Sostarecz, P.A. Janmey, data not published

  5. Texas Red Fluorophore greenred Courtesy of Invitrogen website A. G. Sostarecz, P.A. Janmey, data not published 589 nm615 nm

  6. How are we able to do simultaneous fluorescence?? A. G. Sostarecz, P.A. Janmey, A. Kennedy, data not published

  7. A. G. Sostarecz, P.A. Janmey, A. Kennedy, data not published purple blue greenred Courtesy of Invitrogen website

  8. ELLIPSOMETRY • Determines properties of surfaces and thin films • An ellipsometer allows for the measure of the refractive index and the thickness of semi-transparent thin films. • The reflection at an interface depends on the polarization of the light • The transmission of light through a transparent layer changes the phase of the incoming wave depending on the refractive index of the material • Reflection properties change as the thickness of a sample changes (a few Angstroms)

  9. SET UP – Null Ellipsometer 632.8 nm L = light source (low power HeNe laser) P = polarizing prism Q = quarter-wave plate compensator S = sample (reflects incident light) A = analyzer prism D = light detector

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