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Far-field Subdiffraction Raman Imaging and in situ Correlative Electron Microscopy for Elucidating Details of Plant Cell Wall Structure and Deconstruction

To overcome biomass recalcitrance and harness its potential for energy use, there is a need for in situ characterization of plant cell wall composition, structure, and deconstruction over a range of spatial scales. Raman imaging is an ideal analysis tool for providing both chemical and spatial information.

However, an inherent disadvantage of all traditional far-field optical imaging approaches is limited spatial resolution due to the diffraction of light. To overcome this limitation, we are developing a subdiffraction Raman imaging instrument with 100-nm or better spatial resolution and applying it to study plant cell wall composition, structure, and deconstruction.

This project will deliver a far-field Raman microscope with the capacity to be as versatile and widely applied as any conventional optical microscope. The imaging capabilities will be developed in the context of biomass and bioenergy production from plant cell wall polymers; the impact, however, of the instrument will extend beyond the constraints of this project since the need for chemically specific imaging with nanoscale spatial resolution applies to many biological processes. The proposed studies will enable a better understanding of how plant cell wall properties relate to recalcitrance, and efficient methods for cell wall deconstruction. This will contribute to DOE and BER’s mission to enable “more confident redesign of microbes and plants for sustainable biofuel production.”

This research is supported by the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research (BER) through the Ames Laboratory. The Ames Laboratory is operated for the U.S. Department of Energy by Iowa State University under Contract No. DE-AC02-07CH11358.

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