Micro-sample preparation device for frozen electron microscope and sample preparation method

A technology of cryo-electron microscopy and sample preparation, applied in measuring devices, material analysis using wave/particle radiation, instruments, etc., can solve problems such as large sample consumption, carbon film rupture, and poor volume controllability

Inactive Publication Date: 2021-03-05
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main disadvantage of microfluidic chips used in sample preparation for cryo-electron microscopy is that the continuous injection of sample flow will consume a large amount of sample, with a volume of tens of microliters, and the controllability of the volume of the sample injected on it is poor, and the high-pressure gas will make the copper grid The carbon film on the surface is broken

Method used

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  • Micro-sample preparation device for frozen electron microscope and sample preparation method
  • Micro-sample preparation device for frozen electron microscope and sample preparation method
  • Micro-sample preparation device for frozen electron microscope and sample preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] figure 1 It is a schematic diagram of the structure and sample preparation operation of the device for micro-sample preparation for cryo-electron microscopy in Example 1.

[0068] Sample solution 3 is a ferritin solution, and the sample solution is added to a small centrifuge tube. Using a glass capillary as the sample preparation probe 7, first draw the sample preparation probe 7 in a butane flame to obtain a sample preparation probe 7 with a pointed tip, and then treat the cleaned sample preparation probe 7 with a surface treatment reagent , to obtain the sample preparation probe 7 whose inner and outer surfaces are both hydrophobic. The sample preparation probe 7 uses paraffin oil 15 as a carrier liquid. The sample preparation probe 7 is connected to the liquid driving device 8 composed of a 10 microliter syringe and a precision syringe pump, and then installed on the three-dimensional translation platform 9 to form a liquid droplet manipulation robot. Tweezers ar...

Embodiment 2

[0073] figure 2 It is a schematic diagram of the structure and sample preparation operation of the device for micro-sample preparation for cryo-electron microscopy in Example 2.

[0074] Add sample solution 3 (transferrin solution) to a microcentrifuge tube. A solid metal needle is used as the sample preparation probe 7, the tip of the sample preparation probe 7 is flat, slotted or other shaped structure that is conducive to carrying the sample solution 3, and the region other than the tip of the sample preparation probe 7 is treated with a hydrophobic reagent to obtain A sample preparation probe with a hydrophilic tip7. The sample preparation probe 7 is installed on the three-dimensional translation platform 9 to form a liquid droplet manipulation robot. Tweezers are used as the copper mesh clamping device 5 for fixing the copper mesh 2, and a motor-driven synchronous belt slide table is used as the copper mesh quick transfer device 6. A transparent acrylic plate is used ...

Embodiment 3

[0078] image 3 Schematic diagram of the structure and sample preparation operation of the device for micro-sample preparation for cryo-electron microscopy in Example 3

[0079] Four kinds of different sample solutions 3 to be tested are installed in four holes of a 384-well plate respectively, and ultrapure water is loaded in another hole ( image 3not shown in ). Using a glass capillary as the sample preparation probe 7, first draw the sample preparation probe 7 in a butane flame to obtain a sample preparation probe 7 with a pointed tip, and then treat the cleaned sample preparation probe 7 with a surface treatment reagent , to obtain the sample preparation probe 7 whose inner and outer surfaces are both hydrophobic. The sample preparation probe 7 uses paraffin oil as the carrier liquid. The sample preparation probe 7 is connected to the liquid driving device 8 composed of a 10 microliter syringe and a precision syringe pump, and then installed on the three-dimensional tr...

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Abstract

The invention discloses a micro-sample preparation device for a frozen electron microscope and a sample preparation method. The micro-sample preparation device comprises a supporting net assembly witha sample preparation operation state and a freezing operation state, a sample preparation probe used for respectively carrying out sampling operation and completing sample preparation operation on ato-be-detected sample solution on the supporting net assembly, a three-dimensional translation platform used for driving the sample preparation probe to be converted between the sampling operation andthe sample preparation operation, a freezing module used for carrying out freezing operation on the sample solution on the supporting net assembly, and a rapid transfer device used for driving the supporting net assembly to transfer between the sample preparation operation state and the freezing operation state. According to the invention, a frozen electron microscope sample preparation experiment with ultramicro sample consumption can be realized, and the consumption of a single sample liquid drop is in a nano-liter level and below the nano-liter level. Compared with a conventional sample preparation robot, the sample consumption is reduced by 1000 times or above, the experiment cost is remarkably reduced, and the protein sample purification period is shortened. The success rate of frozen electron microscope sample preparation and the reproducibility of experiments are obviously improved.

Description

technical field [0001] The fields involved in the present invention are the fields of microfluidics, automation and cryo-electron microscopy, and in particular relate to a micro-sample preparation device for cryo-electron microscopy and a sample preparation method thereof. Background technique [0002] Cryo-electron microscopy is currently the mainstream technology for analyzing the structure of biological macromolecules. It takes photos of biological macromolecular particles embedded in amorphous ice embedded in copper mesh carbon membrane holes, and then analyzes and obtains the fine details of biological macromolecules through three-dimensional reconstruction technology. three-dimensional structure. The main process of using cryo-electron microscopy to analyze the three-dimensional structure of proteins and other biomacromolecules includes: sample preparation for cryo-electron microscopy, image acquisition, data processing, and three-dimensional reconstruction. The cryo-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N23/2202G01N23/2204
CPCG01N23/2202G01N23/2204
Inventor 方群王建伟叶升张波张小康张兴
Owner ZHEJIANG UNIV
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