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Shake-it-off: a simple ultrasonic cryo-EM specimen-preparation device
John L Rubinstein1, Hui Guo1, Zev A Ripstein1
1The Hospital for Sick Children Research Institute, Toronto, Ontario, Canada.
Acta Crystallographica. Section D, Structural Biology
|December 4, 2019
Summary
A new device simplifies specimen preparation for electron cryomicroscopy (cryo-EM). Using an ultrasonic humidifier and self-wicking grids, it rapidly creates thin films for high-resolution imaging with minimal sample loss.
Area of Science:
- Structural Biology
- Biophysics
- Microscopy
Background:
- Electron cryomicroscopy (cryo-EM) has advanced significantly, but specimen preparation remains a bottleneck.
- Traditional blotting methods lose over 99.9% of sample and introduce air-water interface artifacts.
- These limitations hinder macromolecular studies and time-resolved experiments.
Purpose of the Study:
- To present a simple, accessible cryo-EM specimen preparation device.
- To enable rapid, low-volume sample application for improved vitrification.
- To facilitate time-resolved cryo-EM studies and reduce sample loss.
Main Methods:
- Utilized components from an ultrasonic humidifier for solution transfer.
- Employed self-wicking electron microscopy (EM) grids with nanowires.
- Controlled the device using a Raspberry Pi single-board computer.
- Designed the device with widely available or manufacturable components.
Main Results:
- Achieved rapid thin film formation within milliseconds of sample application.
- Minimized sample loss compared to traditional blotting techniques.
- Demonstrated a straightforward, open-source design for laboratory construction.
- Enabled customization for specialized cryo-EM experiments.
Conclusions:
- The developed device offers a significant improvement in cryo-EM specimen preparation.
- Its accessibility and simplicity make it suitable for diverse cryo-EM laboratories.
- This method enhances the potential for high-resolution structural determination and time-resolved cryo-EM studies.
