Cryo-EM Grid Sample Spreading With Shock Waves for Uniform Vitrification
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Solution Overview
Problem
Conventional sample preparation techniques for cryo electron microscopy suffer from poor reproducibility and difficulty in achieving the correct thickness of vitrified samples, particularly for single particle analysis and cryo-electron tomography.
Innovation Solution
A method involving the use of shock pressure waves to spread and vitrify a sample solution on a sample holder grid, utilizing a shock pressure wave generator to emit waves at a grazing angle, followed by exposure to a cooling liquid at cryogenic temperatures, with controlled environmental parameters and displacement mechanisms to ensure uniformity and reproducibility.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional manual sample preparation techniques are used, then the process is simple to operate, but the reproducibility and thickness control are poor
Solution Approach 1:
The patent replaces manual mechanical spreading operations with acoustic shock waves generated by a shock wave generator. The shock waves travel through a coupling medium (liquid or gas) to uniformly spread the sample solution across the grid without manual intervention, thereby improving thickness uniformity while reducing operational complexity
Solution Approach 2:
The patent utilizes acoustic pressure waves (a form of pneumatic/hydraulic energy) to drive the sample solution spreading process. The shock waves create pressure variations that propel the sample solution uniformly across the grid surface, achieving consistent thickness control through physical wave mechanics rather than manual manipulation
2Reliability
If manual spreading methods are used, then the equipment is simple, but the reproducibility of sample preparation is poor
Solution Approach 1:
The patent substitutes manual spreading operations with an automated acoustic shock wave system. The shock wave generator produces consistent, repeatable pressure waves that uniformly spread the sample solution, eliminating variability introduced by manual operations and significantly improving preparation reproducibility
Solution Approach 2:
The patent controls critical parameters of the shock wave generation (amplitude, frequency, duration) to ensure consistent and reproducible sample spreading. By precisely adjusting these physical parameters, the system achieves highly reproducible sample preparation results that can be reliably replicated across different experiments
3Manufacturing precision
If rapid cooling is applied to vitrify the sample, then the sample quality is improved, but the control of cooling parameters becomes difficult
Solution Approach 1:
The patent applies the shock waves to spread and position the sample solution uniformly across the grid before initiating the rapid cooling process. This preliminary action ensures that the sample is in the correct configuration and distribution before vitrification, making the subsequent rapid cooling process more controllable and reliable
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
Enables high-quality, reproducible preparation of vitrified samples with uniform thickness, suitable for various types of samples, including proteins and cells, facilitating effective cryo-EM analysis.
Implementation Method 1
spreading the sample solution along the surface of the sample holder grid using one or a sequence of multiple shock pressure waves being generated by a shock pressure wave generator emitting and directing the one or multiple shock pressure waves via at least one nozzle onto the surface of the sample holder grid
Implementation Method 2
vitrifying the spread sample solution by exposing it to a cooling liquid at cryogenic temperatures
Implementation Method 3
may finally be vitrified by rapidly cooling it for example in liquid ethane
Data Source
Figure 1(a)~1(e)
Figure 2(a)~3
Figure 4
AI summary
A sample preparation method and device for preparing a vitrified sample layer (1) on a sample holder grid (3) for subsequent analysis of physical sample characteristics, particularly for subsequent cryo electron microscopy, are presented. The method comprises: providing the sample holder grid (3) with sample solution (5) on at least one surface of the sample holder grid (3); - spreading the dispensed sample solution (5) along the surface (7) of the sample holder grid (3) using one or a sequence of multiple shock pressure waves (9) being generated by a shock pressure wave generator (11) emitting and directing the one or multiple shock pressure waves (9) via at least one nozzle (13) onto the surface (7) of the sample holder grid (3); - vitrifying the spread sample solution (5) by exposing it to a cooling liquid (17) at cryogenic temperatures.