Method for preparing transmission electron microscopy sample of biomimetically synthesized magnetic ferritin nanoparticles

A technology of transmission electron microscope samples and nanoparticles, which is applied in the direction of material analysis, measurement device, and instrument using wave/particle radiation, can solve the problems of difficult preparation, unclean backing of ultra-thin carbon film carrier mesh, low efficiency, etc. Achieve high preparation efficiency, maintain intrinsic dispersion, and eliminate the effects of agglomeration

Active Publication Date: 2019-01-18
INST OF GEOLOGY & GEOPHYSICS CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In view of this, the present invention aims to provide a method for preparing a TEM sample of bionically synthesized magnetic ferritin nanoparticles, which can solve the difficulties, low efficiency, and ultra-thin carbon film loading of magnetic ferritin nanoparticles. The problem of unclean mesh background, and can present the quality of the sample's intrinsic monodispersity

Method used

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  • Method for preparing transmission electron microscopy sample of biomimetically synthesized magnetic ferritin nanoparticles
  • Method for preparing transmission electron microscopy sample of biomimetically synthesized magnetic ferritin nanoparticles
  • Method for preparing transmission electron microscopy sample of biomimetically synthesized magnetic ferritin nanoparticles

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Embodiment 1

[0064] The buffer solution of magnetic ferritin nanoparticles (P-MHFn-5000) obtained by mineralizing recombinant human H subunit ferritin expressed by Pichia pastoris into 5000 iron atoms was determined by BCA kit. Put the amount into a 1.5ml centrifuge tube, add deionized water with a resistivity of 18.2MΩ·cm in proportion, and dilute it to a mixed solution with a concentration of 0.1mg / ml. The solution is slightly yellowish to the naked eye; The mixed solution cooling bath in the tube is ultrasonically oscillated, and ice cubes are added to the ultrasonic wave for cooling, and the ultrasonic time is 10 minutes to obtain a fully mixed sample suspension; select a nickel mesh ultra-thin carbon film carrier grid with a diameter of 3mm and a thickness of <7nm; use The ion fan static eliminator performs electrostatic and dust removal treatment on the ultra-thin carbon film carrier net of the nickel mesh. and time for 10s, then use tweezers to turn the other side of the ultra-thin ...

Embodiment 2

[0071]The magnetic ferritin nanoparticles (E-MHFn-800 ) buffer solution, BCA kit was used for protein concentration determination, M-HFn was diluted to 0.1mg / mL test sample with deionized water with a resistivity of 18.2MΩ·cm; The bath was ultrasonically oscillated, and ice cubes were added to the ultrasonic wave for cooling. The ultrasonic time was 10 minutes to obtain a fully mixed sample suspension; a nickel mesh ultra-thin carbon film carrier grid with a diameter of 3mm and a thickness of <7nm was selected; an ion fan static eliminator was used to The ultra-thin carbon film carrier net of nickel mesh is used for electrostatic dust removal treatment. The ultra-thin carbon film carrier net is placed in a glass culture dish, 30cm away from the ion fan static eliminator, the air volume is controlled at 0.55m3 / min, and the neutralization time is 10s, and then used The tweezers turned the other side of the ultra-thin carbon film carrier grid up, and performed the same operation ...

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Abstract

The invention discloses a method for preparing a transmission electron microscopy sample of biomimetically synthesized magnetic ferritin nanoparticles. The method comprises the following steps: mixinga buffer solution of the biomimetically synthesized magnetic ferritin nanoparticles with deionized water, and diluting to an appropriate concentration; ultrasonically shaking the mixed liquid in a cold bath to obtain a fully mixed sample suspension; performing anti-static dust removal treatment on an ultra-thin carbon film grid; wetting the ultra-thin carbon film grid; placing the treated ultra-thin carbon film grid on clean filter paper, and titrating the sample suspension on the surface of the filter paper; drying the ultra-thin carbon film grid carrying the sample suspension; and after thesample is completely dried, taking the sample out to obtain a magnetic ferritin nanoparticle transmission electron microscopy sample. Through adoption of the method, the problems of difficulty and low efficiency in preparation of the transmission electron microscopy sample of the magnetic ferritin nanoparticles and unclean back bottom of the ultra-thin carbon film grid can be solved, and the intrinsic monodisperse state of the sample can be presented well.

Description

technical field [0001] The invention relates to a method for preparing a transmission electron microscope sample of bionically synthesized magnetic ferritin nanoparticles, belonging to the field of transmission electron microscope sample preparation of ferritin nanoparticles. Background technique [0002] The biomimetic synthesis of magnetic ferritin nanoparticles is a biomineralization protein expressed by genetic engineering—recombinant human H subunit ferritin as a mineralization template. According to the biomineralization principle of ferritin in the human body, a biomimetic synthesis has a typical Magnetic recombinant human ferritin with a core-shell structure, each ferritin shell contains a single crystal nano-magnetite (Fe 3 o 4 ) particles. Biomimetic synthesis of magnetic ferritin nanoparticles usually has the characteristics of monodispersity, uniform shape, controllable particle size, and weak magnetic interaction, and the peroxidase activity of magnetic ferrit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N23/2202
CPCG01N23/2202
Inventor 唐旭
Owner INST OF GEOLOGY & GEOPHYSICS CHINESE ACAD OF SCI
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