Submillisecond second harmonic holographic imaging of biological specimens in three dimensions

Optical microscopy has played a critical role for discovery in biomedical sciences since Hooke’s introduction of the compound microscope. Recent years have witnessed explosive growth in optical microscopy tools and techniques. Information in microscopy is garnered through contrast mechanisms, usuall...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2013-11, Vol.110 (46), p.18391-18396
Main Authors: Smith, David R., Winters, David G., Bartels, Randy A.
Format: Article
Language:English
Subjects:
Animals
Biological activity
Fringe
Geometric shapes
Holography
Holography - instrumentation
Holography - methods
Image reconstruction
Imaging
Laser beams
Mice
Microscopy
Microscopy, Confocal - methods
Muscle, Skeletal - cytology
Optics
Physical Sciences
Scattering
Shot noise
Signal noise
Specimen Handling - methods
Specimens
Supernova remnants
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Summary:Optical microscopy has played a critical role for discovery in biomedical sciences since Hooke’s introduction of the compound microscope. Recent years have witnessed explosive growth in optical microscopy tools and techniques. Information in microscopy is garnered through contrast mechanisms, usually absorption, scattering, or phase shifts introduced by spatial structure in the sample. The emergence of nonlinear optical contrast mechanisms reveals new information from biological specimens. However, the intensity dependence of nonlinear interactions leads to weak signals, preventing the observation of high-speed dynamics in the 3D context of biological samples. Here, we show that for second harmonic generation imaging, we can increase the 3D volume imaging speed from sub-Hertz speeds to rates in excess of 1,500 volumes imaged per second. This transformational capability is possible by exploiting coherent scattering of second harmonic light from an entire specimen volume, enabling new observational capabilities in biological systems.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1306856110