Cryo-EM Grid Transfer Workstation for Ice-Free Lamella Handling
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Solution Overview
Problem
The handling and transfer of electron microscopy grid assemblies during cryo-ET procedures are prone to ice contamination, which obscures the sample and hinders data acquisition, particularly due to the fragile nature of the thin lamellae and the risk of exposing the sample to water during transfer between devices.
Innovation Solution
A workstation with a dry gas environment and overpressure, combined with a transfer device and preparation station, minimizes ice contamination by maintaining a dry gas atmosphere and using cryogenic coolants to keep samples at cryogenic temperatures, allowing for safe handling and transfer of electron microscopy grid assemblies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If manual handling with forceps is used to transfer the electron microscopy grid assembly, then the sample can be transferred between devices, but the fragile lamellae are easily destroyed and ice contamination occurs
Solution Approach 1:
The patent introduces a specialized transfer device with a holder that mechanically grips the grid assembly holder, serving as an intermediary between the operator and the fragile sample. This mediator allows transfer operations without direct manual handling of the lamellae, preventing destruction while enabling movement between devices.
Solution Approach 2:
The patent implements a controlled atmosphere environment (inert or reduced pressure) within the transfer device and workstation. This inert environment prevents ice contamination by excluding water vapor during transfer operations, allowing safe handling without exposure to atmospheric moisture that would otherwise freeze and contaminate the cryogenic sample.
2Ease of operation
If the sample is exposed to atmospheric water during transfer, then transfer between devices can occur, but ice contamination obscures the sample and hinders data acquisition
Solution Approach 1:
The transfer device and workstation maintain an inert or reduced pressure atmosphere that excludes water vapor. This controlled environment prevents atmospheric moisture from contacting the cryogenic sample during transfer, eliminating ice contamination while allowing seamless transfer operations between devices.
Solution Approach 2:
The patent employs sealed chambers and barriers (flexible or rigid enclosures) that create isolated environments during transfer. These physical barriers prevent atmospheric water vapor from reaching the sample, maintaining a contamination-free zone throughout the transfer process.
3Ease of manufacture
If multiple handling steps are performed to prepare and transfer the grid assembly, then comprehensive preparation can be achieved, but the risk of ice contamination and sample damage increases
Solution Approach 1:
The patent combines multiple preparation and transfer operations into an integrated workstation and transfer device system. By merging these functions into a single coordinated system that maintains controlled atmosphere throughout, the number of separate handling steps is reduced, and the sample remains protected from contamination and damage during the entire preparation and transfer sequence.
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
The solution effectively reduces ice contamination, ensuring the integrity of the samples and facilitating seamless transfer and preparation processes without damaging the fragile lamellae, thereby enhancing the quality of cryo-ET data acquisition.
Implementation Method 1
a gas flow of a dry gas, particularly dry nitrogen gas (N2), from a gas reservoir into the first compartment
Implementation Method 2
an overpressure can be generated in the first compartment by the gas flow relative to an exterior of the workstation
Implementation Method 3
transfer the electron microscopy grid assembly to a preparation device, particularly a focused ion beam device used for thinning of the sample, and from the preparation device to the imaging device at cryogenic temperatures
Implementation Method 4
Since the vitrified sample acts as a cold trap, it is especially prone to ice contamination
Data Source
AI summary
The invention relates to a workstation (1), a preparation station (2) and a method for manipulating an electron microscopy grid assembly (3). The workstation (1) comprises a first compartment (101), a first gas inlet (102) for generating an overpressure in the first compartment (101), a first glove (104) and a second glove (105), each being fixed in a respective opening (106, 107) of the workstation (1), wherein the first glove (104) and the second glove (105) are movable in the first compartment (101) to manipulate objects in the first compartment (101), wherein the workstation (1) comprises a port (109) for providing a transfer device (4) for an electron microscopy grid assembly (3) in the first compartment (101). The preparation station (2) comprises a coolant reservoir (201, 202), a first part (210) configured to hold a shuttle (6) for holding an electron microscopy grid assembly (3) in a fixed orientation, wherein the preparation station (2) is configured such that the first part (210) is submergable in the cryogenic coolant when the coolant reservoir (201, 202) contains the cryogenic coolant.