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In Situ Chemical Imaging of Plant Cell Walls Using CARS/SRS Microscopy Yining Zeng 1 , Yu-San Liu 1 , Brian G. Saar 2 , X. Sunney Xie 2 , Fang Chen 3 , Richard A. Dixon 3 , Mike E. Himmel 1 and Shi-You Ding 1 * 1 National Renewable Energy Laboratory, Golden, CO ; 2 Harvard University, Cambridge, MA; 3 The Samuel Roberts Noble Foundation, Ardmore, OK; In collaboration with : David Johnson, Qi Xu (NREL); Gary Holtom, Marcel Friedrich, (Harvard University); Arthur Ragauskas (Georgia Tech); Matthias Hess, Tao Zhang (JGI); Yunfeng Yang, Martin Keller (ORNL) Coherent Anti-Stokes Raman Scattering (CARS ) Imaging of Lignin in Plant Cell Walls Stimulated Raman Scattering (SRS ) • Hand-cut samples; sample treatment is not necessary • Fast imaging rate Simultaneous Chemical Imaging of Lignin and Cellulose During Acidic Pretreatment (Corn Stover ) •CARS intensity reveals lignin content across corn stover cell wall in the order: Epidermis1, sclerenchyma fibers2 > Metaxylem3, Sclerenchyma4, Protoxylem5 >Phloem6 > Parenchyma7 • Selectively chemical bleaching of lignin while cellulose is preserved during the process CARS microscopy provides a powerful tool to monitor real-time and in situ chemical changes during biomass conversion The BioEnergy Science Center (BESC) is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science Selectively chemical bleaching of lignin, while cellulose is preserved during the process. CARS microscopy provides a powerful tool to monitor real-time and in situ chemical changes during biomass conversion. This work is funded by the U.S. Department of Energy Office of Energy Efficiency and Renewable Energy Biomass Program to develop CARS microscopy and image corn stover; the DOE Office of Science, Office of Biological and Environmental Research through the BioEnergy Science Center (BESC) to image transgenic alfalfa and poplar, and “Study of Lignocellulosic Material Degradation with CARS Microscopy” (DE-FG02-07ER64500) to develop new CARS method. Presented at the Bioenergy Sciences Center (BESC) Annual Retreat, 22-24 June 2009, Asheville, North Carolina • NREL/PO-270-46340 NREL is a national laboratory of the U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy, operated by the Alliance for Sustainable Energy, LLC.

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Page 1: Coherent Anti-Stokes Raman Scattering (CARS ) Imaging of ...In Situ Chemical Imaging of Plant Cell Walls Using CARS/SRS Microscopy Yining Zeng1, Yu-San Liu 1, Brian G. Saar2, X. Sunney

In Situ Chemical Imaging of Plant Cell Walls Using CARS/SRS Microscopy

Yining Zeng1, Yu-San Liu1, Brian G. Saar2, X. Sunney Xie2, Fang Chen3, Richard A. Dixon3, Mike E. Himmel1 and Shi-You Ding1* 1National Renewable Energy Laboratory, Golden, CO; 2Harvard University, Cambridge, MA; 3The Samuel Roberts Noble Foundation, Ardmore, OK; In collaboration with: David Johnson,

Qi Xu (NREL); Gary Holtom, Marcel Friedrich, (Harvard University); Arthur Ragauskas (Georgia Tech); Matthias Hess, Tao Zhang (JGI); Yunfeng Yang, Martin Keller (ORNL)

Coherent Anti-Stokes Raman Scattering (CARS ) Imaging of Lignin in Plant Cell Walls

Stimulated Raman Scattering (SRS )

• Hand-cut samples; sample treatment is not necessary• Fast imaging rate

Simultaneous Chemical Imaging of Lignin and Cellulose During Acidic Pretreatment (Corn Stover )

•CARS intensity reveals lignin content across corn stover cell wall in the order: Epidermis1, sclerenchyma fibers2 > Metaxylem3, Sclerenchyma4, Protoxylem5 >Phloem6 > Parenchyma7

• Selectively chemical bleaching of lignin while cellulose is preserved during the process CARS microscopy provides a powerful tool to monitor real-time and in situ chemical changes during biomass conversion

The BioEnergy Science Center (BESC) is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science

Selectively chemical bleaching of lignin, while cellulose is preserved during the process. CARS microscopy provides a powerful tool to monitor real-time and in situ chemical changes during biomass conversion.

This work is funded by the U.S. Department of Energy Office of Energy Efficiency and Renewable Energy Biomass Program to develop CARS microscopy and image cornstover; the DOE Office of Science, Office of Biological and Environmental Research through the BioEnergy Science Center (BESC) to image transgenic alfalfa and poplar,and “Study of Lignocellulosic Material Degradation with CARS Microscopy” (DE-FG02-07ER64500) to develop new CARS method.Presented at the Bioenergy Sciences Center (BESC) Annual Retreat, 22-24 June 2009, Asheville, North Carolina • NREL/PO-270-46340NREL is a national laboratory of the U.S. Department of Energy, Office of Energy Efficiency and Renewable Energy, operated by the Alliance for Sustainable Energy, LLC.

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