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Caenorhabditis elegans fixing method suitable for single-particle microbeam device

A small rod nematode and fixing method technology, applied in the field of radiation biology, to achieve the effects of easy operation, solving high equipment cost and improving recovery rate

Inactive Publication Date: 2015-09-23
AGRI ENG INST ANHUI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Through the self-made simple nematode fixation device, using volatile ethanol and ether mixed anesthetics, and using 2% Agar A gel sheet to moisturize the worm body, it solved the difficulty of nematode fixation and the continuous fixation state during single-particle microbeam irradiation. Existing problems such as short time, slow recovery of worm vigor and low recovery rate after radiation experiments, effectively improve the accuracy and accuracy of single-particle microbeam radiation nematode tissue and organ experiments

Method used

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  • Caenorhabditis elegans fixing method suitable for single-particle microbeam device
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  • Caenorhabditis elegans fixing method suitable for single-particle microbeam device

Examples

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Embodiment 1

[0034] Example 1: Single particle microbeam radiation treatment of L1 / L2 stage larval vulva tissue

[0035]This example describes the use of L1 / L2 stage larvae as an experimental model, using a self-made simple nematode fixation device, using a volatile ethanol and ether mixture for anesthesia and fixation, and then performing single-particle microbeam radiation treatment on the nematode vulva tissue , whose detailed steps are described below.

[0036] Step 1. Cultivation of Caenorhabditis elegans

[0037] Synchronize Caenorhabditis elegans (synchronization reagents: 5 mol / L NaOH and NaClO), place the eggs in 3 ml of M9 buffer and incubate at a constant temperature of 20 degrees Celsius for 24 hours. Observe the nematode eggs under a dissecting microscope. When more than 95% of the eggs hatch into L1 stage larvae, they are inoculated on the NGM nematode solid medium and cultured at a constant temperature of 20 degrees Celsius for 10 hours.

[0038] Step 2. Fabrication of ...

Embodiment 2

[0048] Example 2: Single-particle microbeam irradiation treatment of the retropharyngeal bulb of L4 / early adult worms

[0049] The basic method of this embodiment is the same as that of Example 1, except that in step 1, the synchronized L1 stage larvae are inoculated on NGM medium and cultured for 48 hours. At this time, the nematodes have basically developed to the L4 stage and are about to enter early adulthood; The difference is that in step 4, the L4 stage / early adults need to be anesthetized with a 1:1 mixture of ethanol and ether for 3 minutes, which is due to the strong tolerance of insects at this stage; the difference is also in step 6 It takes 15 minutes for mid-irradiated bugs to fully recover and reach a state of freedom of movement.

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Abstract

The invention discloses a caenorhabditis elegans fixing method suitable for a single-particle microbeam device. The method comprises the main steps of cultivating caenorhabditis elegans, making a simple fixing device, preparing a caenorhabditis elegans body moisturizing gel sheet, fixing the caenorhabditis elegans, preparing radiation samples, and conducting recovery and transfer cultivation on the caenorhabditis elegans after radiation is completed. The caenorhabditis elegans fixing method suitable for the single-particle microbeam device has the advantages that a centrifuge tube sold on the market can be used as the device needed for fixing the caenorhabditis elegans, and the device can be self-made through easy operation; the caenorhabditis elegans fixing process is simple, the consumed time is short, the holding time of the fixed state is long, the recovery method of the caenorhabditis elegans after radiation is simple, the vitality recovery speed is high, and the rate of survival is high. The caenorhabditis elegans body is moisturized through the 2% gel sheet, caenorhabditis elegans dearth caused by rapid water evaporation is effectively avoided, the recovery rate of the caenorhabditis elegans after anesthesia and fixation is increased, the light transmittance of the gel sheet is good, the caenorhabditis elegans body tissue structure under a microscope can be clearly imaged, accurate positioning for single-particle radiation is easy, experiment errors are reduced, and experiment precision is improved.

Description

Technical field: [0001] The invention relates to a method for immobilizing Caenorhabditis elegans, and specifically provides a method for anesthesia of different body tissues of the nematode elegans, which is suitable for fixed-point irradiation of a single-particle microbeam device, and belongs to the technical field of radiation biology. Background technique: [0002] Radiation biology is the study of the interaction of ionizing radiation with cells, tissues and organisms. Traditional radiation biology believes that radiation damage only occurs in directly irradiated cells, tissues or organisms, but the emergence of radiation side effects breaks this traditional view, and has attracted the attention of the majority of researchers. Experimental devices, experimental models and experimental methods for the study of radiation bystander effects. The single-particle microbeam device is widely used in the research of radiation side effects because it can accurately achieve fi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/033
Inventor 郭肖颖施六林王斌
Owner AGRI ENG INST ANHUI ACADEMY OF AGRI SCI
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