NIH%203T3%20fibroblasts%20stained%20for%20Filamentous-actin

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NIH 3T3 fibroblasts stained for
Filamentous-actin Phalloidin Alexa Fluor 488 nm
A. G. Sostarecz, P.A. Janmey, data not published
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Phalloidin Alexa Fluor 488 nm
blue
green
Courtesy of Invitrogen website
A. G. Sostarecz, P.A. Janmey, data not published
E hc ? Longer wavelength lower energy
Alexa Fluor - Fluorescein
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http//probes.invitrogen.com/resources/education/t
utorials/1Introduction/player.html
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210 µm x 170 µm
NIH 3T3 fibroblasts
labeled with fluorescent TR- PIP2
A. G. Sostarecz, P.A. Janmey, data not published
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Texas Red Fluorophore
green red
Courtesy of Invitrogen website
A. G. Sostarecz, P.A. Janmey, data not published
589 nm 615 nm
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How are we able to do simultaneous fluorescence??
A. G. Sostarecz, P.A. Janmey, A. Kennedy, data
not published
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A. G. Sostarecz, P.A. Janmey, A. Kennedy, data
not published
purple
blue
green red
Courtesy of Invitrogen website
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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)

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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