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Improved method for cryopreservation of transmission electron microscope embedding medium

A technology of cryopreservation and transmission electron microscopy, which is applied to the use of radiation for material analysis and preparation of test samples. It can solve the problems of cumbersome sealing steps of tape, bubbles in capsules, and residues of embedding agents, etc., and meet the requirements of clinical diagnosis and treatment. The effect of scientific research needs, less chatter marks, and smooth production process

Pending Publication Date: 2022-04-08
SUZHOU CANCERCELL BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In the prior art, penicillin bottles are used for sub-packaging and cryopreservation, but this method still has the cumbersome steps of sealing with adhesive tape during preservation, the speed of instilling the embedding agent with toothpicks is slow, and the capsules produce more air bubbles after instillation
After the reagent is taken, there are often more embedding agent residues at the bottom of the penicillin bottle, resulting in waste of reagents and other problems

Method used

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  • Improved method for cryopreservation of transmission electron microscope embedding medium
  • Improved method for cryopreservation of transmission electron microscope embedding medium
  • Improved method for cryopreservation of transmission electron microscope embedding medium

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Embodiment 1

[0042] This embodiment provides an improved method for cryopreservation of embedding agents for transmission electron microscopy, the method comprising the following steps:

[0043] (1) Pretreatment of tissue samples:

[0044] Select 30 cases of clinical kidney biopsy specimens, all tissue specimens were cut into 0.1cm×0.1cm×0.1cm in size, placed in 2% glutaraldehyde fixative at 4°C for pre-fixation for 6 hours, 0.2mol phosphate buffer saline Fully rinse at least 3 times, add 1% osmic acid for post-fixation for 1 h; then rinse with buffer solution, and go through gradient dehydration.

[0045] (2) Preparation of embedding agent:

[0046] The embedding agent formula is (mass ratio): epoxy resin EPON618 is 60%, dodecyl succinic anhydride is 25%, methylnadic anhydride is 10%, dibutyl phthalate is 3%, 2.4. 6-tris(dimethylaminomethyl)phenol was 2%.

[0047] After the prepared embedding agent is fully and evenly stirred, take several pre-dried medical disposable 20ml syringes to ...

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Abstract

The invention provides an improved method for cryopreservation of a transmission electron microscope embedding medium. The method comprises the following steps: preparing an embedding medium, extracting the embedding medium by adopting a medical injector, and blocking an opening of the embedding medium; sealing and packaging with a plastic sealing bag, and storing in a freezing chamber of a refrigerator; before use, the frozen embedding medium is taken out to be unfrozen, the embedding medium is dripped into a container by using a medical injector to carry out semi-penetration treatment and full penetration treatment on a tissue specimen, and then the capsule is filled to carry out conventional embedding and polymerization. The embedding block obtained by the preservation method disclosed by the invention is moderate in polymerization hardness and satisfactory in semi-permeation and full-permeation effects; the slide preparation process is smooth, the slice has few wrinkles and tremor marks, the dyeing contrast contrast is obvious, the tissue ultrastructure is clear and distinguishable, the cell structures and substructures are completely preserved, and the dyeing effect is ideal; and when an injector is used for filling the embedding medium into the capsule, the embedding medium can directly reach the bottom of the capsule, and a large number of bubbles generated by a traditional drip irrigation method are avoided.

Description

technical field [0001] The invention belongs to the technical field of medical clinical embedding agents, and relates to an improved method for cryopreservation of embedding agents for transmission electron microscopy. Background technique [0002] Transmission electron microscopy is widely used in ultramicro diagnosis of clinical cases. Obtaining ideal ultra-thin sections is the basis of electron microscope observation, and the rational preparation and use of embedding agent is the key point in basic sample preparation. However, due to the cumbersome preparation procedure of the electron microscope embedding agent and the troublesome measurement of various reagents, a large amount of embedding agent can be prepared at one time and taken in batches as needed, which can adapt to the pace of clinical work, save a lot of time for daily clinical scientific research work, and greatly improve the efficiency of clinical research. Improve work efficiency and reduce labor intensity....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/36G01N1/42G01N23/04
Inventor 王守立彭金玉吴月
Owner SUZHOU CANCERCELL BIOTECH
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