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Fabrication of Micro-Patterned Chip with Controlled Thickness for High-Throughput Cryogenic Electron Microscopy
Published on: April 21, 2022
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Fabrication of Micro-Patterned Chip with Controlled Thickness for High-Throughput Cryogenic Electron Microscopy
Min-Ho Kang1, Minyoung Lee2, Sungsu Kang2
1Department of Biomedical-Chemical Engineering, The Catholic University of Korea; Department of Biotechnology, The Catholic University of Korea.
Journal of Visualized Experiments : Jove
|May 9, 2022
Summary
Researchers developed a novel silicon chip with graphene oxide windows for improved cryogenic electron microscopy (cryo-EM) sample preparation. This technique allows controlled ice thickness, enhancing high-throughput analysis of biomolecules and nanomaterials.
Area of Science:
- Materials Science
- Biophysics
- Nanotechnology
Background:
- Cryogenic electron microscopy (cryo-EM) demands precise nanoscale ice thickness for high-throughput structural analysis.
- Current methods for cryo-EM sample preparation face limitations in controlling ice thickness efficiently.
- Developing advanced sample carriers is crucial for advancing biomolecular structure determination.
Purpose of the Study:
- To engineer a novel micro-patterned chip for on-demand control of ice thickness in cryo-EM sample preparation.
- To enhance the efficiency and throughput of structural analysis for biomolecules and nanomaterials using cryo-EM.
- To leverage microelectromechanical system (MEMS) techniques and graphene oxide (GO) for improved cryo-EM sample quality.
Main Methods:
- Fabrication of silicon-based chips using microelectromechanical system (MEMS) techniques, including UV photolithography and etching.
- Patterning of graphene oxide (GO) windows on thickness-controlled silicon nitride (SixNy) films.
- Utilizing the affinity of GO for biomolecules to concentrate samples within micro-holes for cryo-EM analysis.
Main Results:
- Successful mass-production of micro-patterned chips with GO windows.
- Demonstrated on-demand control of ice thickness by regulating micro-hole depth for various specimen sizes.
- Observed concentration of biomolecules within micro-holes due to GO's favorable affinity.
- Enabled high-throughput cryo-EM imaging of diverse biological molecules and inorganic nanomaterials.
Conclusions:
- The developed micro-patterned chip with GO windows significantly overcomes limitations in cryo-EM sample preparation.
- This innovation facilitates efficient, high-throughput structural analysis of biomolecules and nanomaterials.
- The chip represents a significant advancement for cryo-EM applications in structural biology and materials science.
