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Cryo-FIB & Cryo-ET: 3D Visualisation of Cellulose and Meshing Architecture in Onion Cell Walls


This video offers an unprecedented look into the nanoscale architecture of the plant cell wall, demonstrating the power of correlative cryo-microscopy techniques. The goal is to overcome the challenge of visualising the cell wall's dense, complex structure in its native, hydrated state.


Methodology Featured:

We first utilize Cryo-FIB (Focused Ion Beam) Milling to meticulously shave away material from the frozen, bulky onion cell wall peels. This essential step generates ultra-thin, electron-transparent lamellae, which are perfectly suited for high-resolution imaging.


Subsequently, we perform Cryo-ET (Cryo-Electron Tomography) on these lamellae. By acquiring multiple images at different tilt angles, we computationally reconstruct a 3D tomogram, revealing the intricate organisation of the cell wall components.


Key Visualisations:

The video visually highlights the structural findings:


Cellulose Fibers (Yellow): Observe the rigid, load-bearing microfibrils that give the wall its strength and directionality.


Meshing (Red): Visualize the less-ordered, potentially pectin-rich matrix material that forms a network (meshing) around the cellulose scaffold.


This demonstration is a valuable resource for researchers and students in plant biology, structural biology, and biophysics, showcasing the cutting-edge methods used to study native cellular structures at molecular resolution.


Keywords: Cryo-FIB Milling, Cryo-ET, Cryo-Electron Tomography, Onion Cell Wall, Onion, Pectins


Citation:

Nicolas, W. J., Fäßler, F., Dutka, P., Schur, F. K., Jensen, G., & Meyerowitz, E. (2022). Cryo-electron tomography of the onion cell wall shows bimodally oriented cellulose fibers and reticulated homogalacturonan networks. Current Biology, 32(11), 2375-2389.


Attribution 4.0 International — CC BY 4.0 - Creative Commons


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