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Non-Linear Contrast Mechanisms for Optical Microscopy

  • Virginijus Barzda
Part of the Advances in Photosynthesis and Respiration book series (AIPH, volume 26)

Optical microscopy is a primary research tool that is widely used for visualization of cells and subcellular organelles in living photosynthetic organisms. Although being primarily an imaging tool, a microscope can also be used as a spectroscopic instrument that offers means to access spectral properties of individual photosynthetic membranes and pigment-protein complexes with a diffraction limited spatial resolution. When coupled with a femtosecond laser, the optical microscope becomes an imaging tool that differentiates the biological structures based on non-linear light-matter interactions. A large plethora of non-linear optical responses can be employed for imaging, and spatially resolved spectroscopy, rendering structural and functional information about photosynthetic systems.

This chapter will introduce three non-linear contrast mechanisms that have been applied for microscopic investigations of photosynthetic samples: second harmonic generation, third-harmonic generation, and multiphoton excitation fl uorescence. Two applications of non-linear microscopy will be presented featuring structural and spectroscopic investigations of individual in situ chloroplasts and isolated aggregates of plant major lightharvesting chlorophyll a/b pigment-protein complexes associated with Photosystem II ( LHCII). Readers will be exposed to novel tools and possibilities of the rapidly growing fi eld of non-linear microscopy.

Keywords

Harmonic Generation Third Harmonic Generation Nonlinear Microscopy Photosynthetic Sample Harmonic Generation Microscopy 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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© Springer Science+Business Media B.V 2008

Authors and Affiliations

  • Virginijus Barzda
    • 1
  1. 1.Department of Physics, and Institute for Optical SciencesUniversity of TorontoMississaugaCanada

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