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Cell Culture on Silicon Nitride Membranes and Cryopreparation for Synchrotron X-ray Fluorescence Nano-analysis
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Biochemistry
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JoVE Journal Biochemistry
Cell Culture on Silicon Nitride Membranes and Cryopreparation for Synchrotron X-ray Fluorescence Nano-analysis

Cell Culture on Silicon Nitride Membranes and Cryopreparation for Synchrotron X-ray Fluorescence Nano-analysis

DOI:
10.3791/60461-v

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08:26 min

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December 10, 2019

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, , , , , ,
1Inserm, UA7, Synchrotron Radiation for Biomedicine (STROBE),Université Grenoble Alpes, 2ID16A Beamline,ESRF, The European Synchrotron

Chapters

  • 00:04Title
  • 00:59Silicon Nitride (Si3N4) Membrane Support Preparation
  • 02:14Cell Seeding
  • 03:04Plunge-Freezing
  • 04:46Plunge-Frozen Cell Freeze-Drying
  • 06:05Results: Representative X-Ray Fluorescence Imaging of Frozen Hydrated and Freeze-Dried Cells
  • 07:28Conclusion

Summary

Automatic Translation

Automatic Translation

Presented here is a protocol for cell culture on silicon nitride membranes and plunge-freezing prior to X-ray fluorescence imaging with a synchrotron cryogenic X-ray nanoprobe. When only room temperature nano-analysis is provided, the frozen samples can be further freeze-dried. These are critical steps to obtain information on the intracellular elemental composition.

Tags

Keywords: Cell CultureSilicon Nitride MembraneCryopreparationSynchrotron X-ray Fluorescence Nano-analysisCryofixationFrozen Hydrated CellsElemental DistributionRadiation DamageCell MonolayerPoly-L-lysineAttachment FactorUltra Pure WaterCell SeedingCell Culture Medium

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