Optical microscopy in photosynthesis.
Emerging as well as the most frequently used optical microscopy techniques are reviewed and image contrast generation methods in a microscope are presented, focusing on the nonlinear contrasts such as harmonic generation and multiphoton excitation fluorescence. Nonlinear microscopy presents numerous advantages over linear microscopy techniques including improved deep tissue imaging, optical sectioning, and imaging of live unstained samples. Nonetheless, with the exception of multiphoton excitation fluorescence, nonlinear microscopy is in its infancy, lacking protocols, users and applications; hence, this review focuses on the potential of nonlinear microscopy for studying photosynthetic organisms. Examples of nonlinear microscopic imaging are presented including isolated light-harvesting antenna complexes from higher plants, starch granules, chloroplasts, unicellular alga Chlamydomonas reinhardtii, and cyanobacteria Leptolyngbya sp. and Anabaena sp. While focusing on nonlinear microscopy techniques, second and third harmonic generation and multiphoton excitation fluorescence microscopy, other emerging nonlinear imaging modalities are described and several linear optical microscopy techniques are reviewed in order to clearly describe their capabilities and to highlight the advantages of nonlinear microscopy.
Department of Chemical and Physical Sciences, University of Toronto, 3359 Mississauga Road, Mississauga, ON, L5L 1C6, Canada.
Richard
R
Cisek
Leigh
L
Spencer
Nicole
N
Prent
Donatas
D
Zigmantas
George S
GS
Espie
Virginijus
V
Barzda
D008853:Microscopy / Q000379:methods*; D017711:Nonlinear Dynamics; D055620:Optical Phenomena; D010788:Photosynthesis / Q000502:physiology*