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Treatment method of micro organ

A treatment method and organ technology, applied in the preparation of test samples, etc., can solve problems such as high requirements for research conditions, difficulties in observation, and obstacles to research, and achieve the effect of high proficiency requirements, strong comparability, and easy success

Inactive Publication Date: 2018-01-09
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Usually, the organs of tiny animals are very small, and the original tissue anatomy research conditions are demanding and difficult, especially some with a thickness of less than 1mm, which are difficult to observe with the naked eye, and the outermost layer is non-keratinized or ossified. Internal micro-organs whose reflection contrast is very small and whose visibility is close to 0 (for example, unlike external tissues and organs such as the body wall and head of silkworm larvae, in order to resist damage to internal tissues by sharp objects, the outermost layer is keratinized and ossified , after transparent treatment, it still retains a certain light reflection contrast and visibility), when scientific research is carried out at the molecular level, because there is no suitable and feasible method, it seriously hinders related research and becomes a bottleneck for in-depth research.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Immune Response of Internal Miniature Organs of Bombyx mori Larvae

[0033] A processing method for micro-organs, comprising the following steps:

[0034] (1) Dissect the silkworm larvae, extract the micro-organs of the purpose, and remove other sundries adhering to them in PBS;

[0035] (2) The micro-organs were treated in two stages with the treatment liquid, soaked in light shaking for 4 minutes in each stage, the formula of the treatment liquid was 10 μL Tween-20, 10 mL 0.01M PBS;

[0036] (3) Fix the micro-organ with 4% paraformaldehyde (dissolved in 0.1M PBS) at 4°C for 14h;

[0037] (4) The micro-organs were treated in two stages with the treatment liquid, soaked in light shaking for 12 minutes in each stage, the formula of the treatment liquid was 10 μL Tween-20, 10 mL 0.01M PBS;

[0038] (5) Dialyze the micro-organs in dialysate I at 4°C for 4 days; the formula of dialysate I: 1M Tris 15mL, NaCl 2.63g, Triton-X 1001.5mL, BSA 0.3g, pH 6.8;

[0039] ...

Embodiment 2

[0053] Example 2 Cytological analysis of silkworm larva pharyngeal side body

[0054] A processing method for micro-organs, comprising the following steps:

[0055] (1) Dissect the silkworm larvae, extract the brain-cardiolateral body-pharynx lateral body complex, and remove other debris adhering to it in PBS;

[0056] (2) The micro-organs were treated in two stages with the treatment liquid, soaked in light shaking for 5 minutes in each stage, the formula of the treatment liquid was 10 μL Tween-20, 10 mL 0.01M PBS;

[0057] (3) Fix the micro-organ with 4% paraformaldehyde (dissolved in 0.1M PBS) at 4°C for 15h;

[0058] (4) The micro-organs were treated in two stages with the treatment liquid, soaked in light shaking for 15 minutes in each stage, and the formula of the treatment liquid was 10 μL Tween-20, 10 mL 0.01M PBS;

[0059] (5) Dialyze the micro-organs in dialysate I at 4°C for 3 days; the formula of dialysate I: 1M Tris 15mL, NaCl 2.63g, Triton-X 1001.5mL, BSA 0.3g,...

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Abstract

The invention provides a treatment method of a micro organ. The treatment method comprises the following step sequentially dialyzing a fixed micro organ in dialysate I for 2 to 5 days under 1 to 10 DEG C and in dialysate II for 4 to 8 hours under -40 to -15 DEG C, wherein a formula of the dialysate I comprises 1M of Tris alkaline solution which is prepared from 10 percent of Triton-X 100, 2 percent of BSA (Bull Serum Albumin) and 0.05M of NaCl, and the pH (Potential of Hydrogen) is 6.8; a formula of the dialysate II comprises ethanol and DMSO (Dimethyl Sulfoxide) which are 4 to 1 in volume ratio. The treatment method provided by the invention is used for carrying out sizing and whole permeation treatment on the micro organ, an original morphology of the micro organ is kept, and the purposeof enabling macromolecular matters to fully permeate each position without changing the structural morphology of the micro organ and tissue cells can be realized; the treated micro organ is capable of accurately and sensitively carrying out cellular or subcellular location and qualitative and relatively quantitative analysis on molecules on the micro organ by utilizing specific binding reaction of molecular matters and through corresponding observation means such as a fluorescent microscope. Aiming at the difficulties that internal organs of animalcules in the field of biology are very smalland a tissue section technique is difficult to implement for the internal organs, the method for researching in-vivo micro organ on molecular and cellular level is invented.

Description

technical field [0001] The invention relates to a method for treating tissues and organs, in particular to tiny tissues and organs in animals. technical background [0002] The animal body is a combination of various tissues and organs, and the life activities of animals must be maintained by the normal performance of their respective physiological and biological functions by each tissue and organ. The study of tissues and organs is the premise and important link for in-depth understanding of animals and the control and utilization of animals. [0003] Histochemistry, a commonly used tissue and organ pathological detection method in medical diagnosis, has been sublimated into immunohistochemistry in the biological field with the development of molecular and cell biology. Immunohistochemistry skillfully combines the immunoreaction specificity of antigen-antibody binding with the visibility of histochemistry, detects antigenic substances at the cellular and subcellular levels...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30G01N1/28
Inventor 陈萍李田张亮陈曦李亚腾
Owner SOUTHWEST UNIV
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