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.


Harmonic Generation Third Harmonic Generation Nonlinear Microscopy Photosynthetic Sample Harmonic Generation Microscopy 
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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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