Huimin Xu1,2,3,4, Yuanyuan Zhao2,3,5, Yuanzhen Suo4,6, Yayu Guo2,3,5, Yi Man2,3,5, Yanping Jing2,3,5, Xinqiang He4, Jinxing Lin7,8,9. 1. College of Biological Sciences, China Agricultural University, Beijing, 100193, China. 2. Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, Beijing Forestry University, Beijing, 10083, China. 3. College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, 100083, China. 4. School of Life Sciences, Peking University, Beijing, 100871, China. 5. Institute of Tree Development and Genome Editing, Beijing Forestry University, Beijing, 100083, China. 6. Biomedical Pioneering Innovation Center, Peking University, Beijing, 100871, China. 7. Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, Beijing Forestry University, Beijing, 10083, China. linjx@ibcas.ac.cn. 8. College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, 100083, China. linjx@ibcas.ac.cn. 9. Institute of Tree Development and Genome Editing, Beijing Forestry University, Beijing, 100083, China. linjx@ibcas.ac.cn.
Abstract
BACKGROUND: New cell wall imaging tools permit direct visualization of the molecular architecture of cell walls and provide detailed chemical information on wall polymers, which will aid efforts to use these polymers in multiple applications; however, detailed imaging and quantification of the native composition and architecture in the cell wall remains challenging. RESULTS: Here, we describe a label-free imaging technology, coherent Raman scattering (CRS) microscopy, including coherent anti-Stokes Raman scattering (CARS) microscopy and stimulated Raman scattering (SRS) microscopy, which can be used to visualize the major structures and chemical composition of plant cell walls. We outline the major steps of the procedure, including sample preparation, setting the mapping parameters, analysis of spectral data, and image generation. Applying this rapid approach will help researchers understand the highly heterogeneous structures and organization of plant cell walls. CONCLUSIONS: This method can potentially be incorporated into label-free microanalyses of plant cell wall chemical composition based on the in situ vibrations of molecules.
BACKGROUND: New cell wall imaging tools permit direct visualization of the molecular architecture of cell walls and provide detailed chemical information on wall polymers, which will aid efforts to use these polymers in multiple applications; however, detailed imaging and quantification of the native composition and architecture in the cell wall remains challenging. RESULTS: Here, we describe a label-free imaging technology, coherent Raman scattering (CRS) microscopy, including coherent anti-Stokes Raman scattering (CARS) microscopy and stimulated Raman scattering (SRS) microscopy, which can be used to visualize the major structures and chemical composition of plant cell walls. We outline the major steps of the procedure, including sample preparation, setting the mapping parameters, analysis of spectral data, and image generation. Applying this rapid approach will help researchers understand the highly heterogeneous structures and organization of plant cell walls. CONCLUSIONS: This method can potentially be incorporated into label-free microanalyses of plant cell wall chemical composition based on the in situ vibrations of molecules.