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Microfluidic Liquid Cell with Silicon Nitride Super-Thin Membrane for Electron Microscopy of Samples in Liquid
Microfluidic liquid cells have been developed to visualize nanoscaled biological samples in liquid using a scanning electron microscope (SEM) through an electron-transparent membrane (ETM). However, despite the combination of the high-resolution visualization of SEM and the high experimental capabil...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9775586/ https://www.ncbi.nlm.nih.gov/pubmed/36551105 http://dx.doi.org/10.3390/bios12121138 |
Sumario: | Microfluidic liquid cells have been developed to visualize nanoscaled biological samples in liquid using a scanning electron microscope (SEM) through an electron-transparent membrane (ETM). However, despite the combination of the high-resolution visualization of SEM and the high experimental capability of microfluidics, the image is unclear because of the scattering of the electron beam in the ETM. Thus, this study developed a microfluidic liquid cell with a super-thin ETM of thickness 10 nm. Because the super-thin ETM is excessively fragile, the bonding of a silicon–nitride-deposited substrate and a polydimethylsiloxane microchannel before silicon anisotropic etching was proposed prevented the super-thin ETM from damage and breakage due to etching. With this protection against etchant using the microchannel, the yield of the fabricated super-thin ETM increased from 0 to 87%. Further, the scattering of the electron beam was suppressed using a microfluidic liquid cell with a super-thin ETM, resulting in high-resolution visualization. In addition, T4 bacteriophages were visualized using a super-thin ETM in vacuum. Furthermore, the cyanobacterium Synechocystis sp. PCC6803 in liquid was visualized using a super-thin ETM, and sub-microscopic structures on the surface were observed. |
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