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Methods and devices for preparing sample for cryogenic electron microscopy

a cryogenic electron microscopy and sample technology, applied in material analysis, material analysis using wave/particle radiation, instruments, etc., can solve the problems of trial and error testing, extremely time-consuming,

Active Publication Date: 2021-03-11
FEI CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

The patent describes a device used to inspect samples for their condition. The device has multiple chambers and pillars that hold samples. Each sample is inspected through a window in the device. This allows for screening multiple sample conditions within the same device.

Problems solved by technology

However, preparing and imaging vitrified protein samples with different sample conditions requires extensive trial and error testing, and can become extremely time consuming.

Method used

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  • Methods and devices for preparing sample for cryogenic electron microscopy
  • Methods and devices for preparing sample for cryogenic electron microscopy
  • Methods and devices for preparing sample for cryogenic electron microscopy

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Embodiment Construction

[0018]The following description relates to systems and methods for screening samples for cryogenic electron microscopy (cryo-EM) based single particle analysis. The cryo-EM imaging may be performed using an imaging system shown in FIG. 1.

[0019]In order to perform single particle analysis, the sample is undergone multiple sample preparation steps to achieve an optimal sample condition before being imaged with the cryo-EM imaging system. The sample condition may include buffer type and PH level, salt and detergent type and concentration, as well as surfactant concentration. The sample condition may also include nanodisc size, lipid to protein ratio, and critical micelle concentration.

[0020]The sample preparation steps may include purifying the sample, mixing the purified sample with buffers, and vitrifying the mixture. One of the sample preparation steps is mixing the sample, such as a protein, with one or more buffers to solubilize and stabilize the sample. For example, the protein m...

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Abstract

Various methods and devices are provided for searching the optimum sample condition of a sample for cryogenic electron microscopy. Multiple samples with different sample conditions may be screened using a sample inspection device. The sample inspection device includes at least a chamber formed between a top electron transparent layer and a bottom electron transparent layer for holding the sample. Multiple pillars are arranged within the chamber. The sample inspection device includes a window covering at least one of the multiple pillars.

Description

FIELD OF THE INVENTION[0001]The present description relates generally to methods and devices for preparing samples for single particle analysis, and more particularly, to optimizing the sample quality for single particle analysis based on cryogenic electron microscopy imaging.BACKGROUND OF THE INVENTION[0002]Single particle analysis based on cryogenic electron microscopy (cryo-EM) images may solve structures of biological macromolecules at near atomic resolution. Sample preparation is a key step for obtaining high quality cryo-EM images of the macromolecules. As an example, in order to solve protein structure, the protein sample is prepared by mixing the protein with one or more buffer solutions. The sample construct, as well as buffer conditions including PH level, salt concentration, surfactant concentration, and detergent concentration, need to be optimized to solubilize and stabilize the protein for the optimal cryo-EM result. However, preparing and imaging vitrified protein sam...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N23/225G01N1/42G01N23/2204
CPCG01N23/225H01J2237/221G01N23/2204G01N1/42G01N23/04G01N23/20058G01N1/38G01N2001/386H01J2237/2004H01J37/20G01N23/2202G01N2223/418G01N2223/307
Inventor HENDRIKSEN, BASKUIJPER, MAARTENMELE, LUIGIDONA, PLEUNRAJA, ERUMAMINIAN, ATIEH
Owner FEI CO
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