Cryogenic Sample Vitrification Using Direct Liquid Coolant Jets
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Solution Overview
Problem
Current methods for preparing samples under cryogenic conditions for imaging or diffraction experiments often result in artifacts due to the reinforcement features of the sample support, and fail to achieve extreme cooling rates necessary for optimal vitrification.
Innovation Solution
A method and apparatus that vitrifies samples by directing a jet of liquid coolant directly onto the sample, achieving cooling rates exceeding 50,000 K/s, and uses synchronized jets and a plasma treatment to prevent artifact formation and ensure even sample application on a hydrophilic support.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional plunge freezing methods are used, then samples can be cooled to cryogenic temperatures, but artifact formation occurs due to support reinforcement features and insufficient cooling rates
Solution Approach 1:
The invention extracts the harmful reinforcement features from the sample support by using a support without reinforcement features during the critical vitrification process. The support is designed to be simple and uniform in the region where samples are applied, eliminating the source of artifacts that would otherwise be present in conventional supported films.
Solution Approach 2:
The invention applies preliminary action by pre-cooling the support carrier to cryogenic temperature before applying the liquid sample. This ensures that the support is already at the required temperature to immediately begin the vitrification process upon sample application, achieving the extreme cooling rates necessary for high-quality vitrification without artifacts.
2Temperature
If conventional cooling rates are used, then samples can be frozen, but extreme cooling rates exceeding 50,000 K/s are not achieved, resulting in poor vitrification
Solution Approach 1:
The invention uses a hydraulic or pneumatic actuation system to rapidly eject the support carrier from the support preparation device into the cryogenic environment. This rapid ejection mechanism achieves the extreme cooling rates exceeding 50,000 K/s necessary for optimal vitrification by minimizing the time at intermediate temperatures where ice crystal formation could occur.
3Strength
If samples are applied to conventional supports, then samples can be supported for imaging, but reinforcement features of the support cause artifacts in images
Solution Approach 1:
The invention applies local quality by providing a support with different properties in different regions: the region where samples are applied has no reinforcement features to avoid artifacts, while other regions of the support may have reinforcement features if needed for mechanical handling. This localized differentiation ensures that samples are imaged over a uniform, artifact-free region while the support maintains sufficient overall strength.
4Productivity
If automated sample preparation is implemented, then productivity and consistency improve, but device complexity increases
Solution Approach 1:
The invention implements a multi-functional apparatus that combines sample application, support cooling, and rapid ejection mechanisms into a single integrated device. This universal device performs multiple functions (pre-cooling the support, applying the liquid sample, and rapidly ejecting the support into the cryogen) in sequence, improving productivity and consistency while managing device complexity through functional integration rather than separate components.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach effectively vitrifies samples with extreme cooling rates, reducing the impact of support features and preventing artifact formation, while allowing for precise control of sample preparation and handling.
Implementation Method 1
directing a jet of liquid coolant directly onto the sample, achieving cooling rates exceeding 50,000 K/s
Implementation Method 2
uses synchronized jets and a plasma treatment to prevent artifact formation and ensure even sample application on a hydrophilic support
Implementation Method 3
vitrifies samples by directing a jet of liquid coolant directly onto the sample
Data Source
AI summary
The invention relates to a method of and apparatus for preparing a sample for imaging or diffraction experiments under cryogenic conditions, comprising the steps of applying a sample to sample carrier, such as a film on a support, in particular a grid comprising such a film on a support, or removing residual medium, typically liquid, from an incubated sample on a film on a support, and vitrifying the sample. The sample is vitrified by directing a jet of liquid coolant to the center of the film and onto the sample.