Method for marking gastrointestinal tract of zebra fish with calcein

A calcein, gastrointestinal technology, applied in the field of drug screening, can solve problems such as long experimental period, achieve the effects of intuitive observation, enhanced peristalsis and feeding ability, and improved accuracy and reliability

Active Publication Date: 2016-06-29
BIOLOGY INST OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preferred marking time of this method is 16h, and the whole experiment period is longer

Method used

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  • Method for marking gastrointestinal tract of zebra fish with calcein
  • Method for marking gastrointestinal tract of zebra fish with calcein
  • Method for marking gastrointestinal tract of zebra fish with calcein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Labeling the gastrointestinal tract of zebrafish with different concentrations of calcein

[0056] 1. Zebrafish sample pretreatment:

[0057] Use healthy and sexually mature AB zebrafish, put them into a mating tank at a ratio of 1:1 or 1:2, place a partition in the middle, and place them in a dark environment. Remove the partition before turning on the light the next day, and light stimulate Make it ovulate, fish out adult fish after ovulation half an hour, control the ovulation time within half an hour, to reduce the difference in development time between embryos. Collect the fertilized eggs and rinse them three times with new zebrafish embryo culture water to disinfect and clean the fertilized eggs; then transfer the fertilized eggs into clean water for zebrafish embryo culture, and control the temperature at 28°C with a 14h light / 10h dark cycle. Culture in light, change about 1 / 3 of the water every 24 hours, and suck out the dead embryos in time. The 6-...

Embodiment 2

[0068] Example 2: Calcein with a mass concentration of 0.2% labels the gastrointestinal tract of zebrafish at different temperatures

[0069] 1. Zebrafish sample pretreatment:

[0070] Use healthy and sexually mature AB zebrafish, put them into a mating tank at a ratio of 1:1 or 1:2, place a partition in the middle, and place them in a dark environment. Remove the partition before turning on the light the next day, and light stimulate Make it ovulate, fish out adult fish after ovulation half an hour, control the ovulation time within half an hour, to reduce the difference in development time between embryos. Collect the fertilized eggs and rinse them three times with new zebrafish embryo culture water to disinfect and clean the fertilized eggs; then transfer the fertilized eggs into clean water for zebrafish embryo culture, and control the temperature at 28°C with a 14h light / 10h dark cycle. Culture in light, change about 1 / 3 of the water every 24 hours, and suck out the dead...

Embodiment 3

[0081] Example 3: Calcein with a mass concentration of 0.2% labels the gastrointestinal tract of zebrafish at different times

[0082] 1. Obtaining and using zebrafish embryos

[0083] Use healthy and sexually mature AB zebrafish, put them into a mating tank at a ratio of 1:1 or 1:2, place a partition in the middle, and place them in a dark environment. Remove the partition before turning on the light the next day, and light stimulate Make it ovulate, fish out adult fish after ovulation half an hour, control the ovulation time within half an hour, to reduce the difference in development time between embryos. Collect the fertilized eggs and rinse them three times with new zebrafish embryo culture water to disinfect and clean the fertilized eggs; then transfer the fertilized eggs to clean water for zebrafish embryo culture, at 28°C, 14h light / 10h dark cycle under light control During cultivation, about 1 / 3 of the water was changed every 24 hours, and dead embryos were sucked ou...

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Abstract

The invention relates to a method for marking a gastrointestinal tract of zebra fish with calcein. The method comprises the following steps of: (1) selecting normally developed zebra fish embryos, putting the zebra fish embryos into a culture vessel containing a culture solution and adding a calcein solution; (2) incubating the zebra fish embryos under a constant-temperature condition in a shading condition, cleaning the zebra fish embryos with the culture solution and preparing marked the zebra fish embryos; and (3) narcotizing the marked the zebra fish embryos, and carrying out image collection and storage on the gastrointestinal tract part for displaying yellow-green fluorescent lights in the zebra fish embryos by an image collection tool. The influence factors for the gastrointestinal tract of the zebra fish marked with the calcein are researched for the first time; the condition that the influence factors for the gastrointestinal tract of the zebra fish marked with the calcein are different from the influence factors for dyeing in manners of skeleton deposition and the like is found out; after related conditions are optimized, the marking method capable of obtaining the result which is clear in gastrointestinal tract outline, clear, visible and easy to identify can be obtained.

Description

technical field [0001] The invention relates to a method for marking the gastrointestinal tract of zebrafish with calcein, belonging to the technical field of drug screening. Background technique [0002] The gastrointestinal tract is the largest immune organ in the human body, and it is also the largest detoxification organ in the human body. The gastrointestinal tract refers to the digestive tube that runs from the stomach pylorus to the anus. The intestine is the longest section of the alimentary canal and the most functionally important section. Since the reform and opening up, with the rapid development of the economy, the increasingly fierce social competition, the acceleration of people's life rhythm and the increase of work pressure, the problems of gastrointestinal diseases have become more and more common. So far, this type of disease has become a common clinical disease and frequently-occurring disease, and the total incidence accounts for about 20% of the total...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00
CPCA61K49/0008A61K49/0043A61K49/006
Inventor 孙晨韩建韩利文王雪王荣春侯海荣陈锡强
Owner BIOLOGY INST OF SHANDONG ACAD OF SCI
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