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VitriFlex: An Open-Source, Modular, and Customizable Robotic Platform for Cryo-EM Grid Preparation
Wyatt Peele1, Kedar Sharma1, Thomas B Stanley1
1Genome Integrity & Structural Biology Laboratory National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services. Research Triangle Park, NC, US.
Biorxiv : the Preprint Server for Biology
|April 17, 2026
Summary
VitriFlex is an open-source platform for cryo-electron microscopy (cryo-EM) specimen preparation. This automated system offers modularity and programmable control, enabling high-resolution cryo-EM data acquisition for various applications.
Area of Science:
- Structural Biology
- Biophysics
- Microscopy
Background:
- Specimen preparation is a major challenge in cryo-electron microscopy (cryo-EM), impacting data quality.
- Existing commercial systems are often closed, expensive, and lack experimental flexibility, especially for time-resolved studies.
- There is a need for adaptable, accessible platforms for cryo-EM grid preparation.
Purpose of the Study:
- To develop an open-source, modular, and automated platform for cryo-EM specimen preparation.
- To overcome limitations of current commercial plungers, particularly for time-resolved experiments and small sample volumes.
- To enable researchers to customize and optimize grid preparation parameters.
Main Methods:
- Developed VitriFlex, an open-source platform using a SCARA robot and 3D-printed components.
- Implemented programmable control for grid handling, sample deposition (including acoustic-assisted spray), and blotting.
- Enabled flexible sample delivery strategies like pre-mixing and on-grid mixing.
Main Results:
- VitriFlex reproducibly produced high-quality, collectable grids for standard samples (apoferritin, dGTPase) and complex mixtures (spike-ACE2, α7-bungarotoxin).
- Achieved high-resolution reconstructions from VitriFlex-prepared grids.
- Demonstrated short spray-to-plunge delays (∼130 ms) for time-resolved biochemical reactions.
Conclusions:
- VitriFlex provides a flexible, accessible, and customizable solution for routine and time-resolved cryo-EM.
- The open-source nature lowers adoption barriers and facilitates integration of new components.
- This platform advances cryo-EM specimen preparation, enabling high-resolution structural studies.
