Automatic building device of transgenic fish

An automatic and control device technology, applied in genetic engineering, biochemical cleaning devices, enzymology/microbiology devices, etc., can solve problems such as low efficiency, cumbersome operation, and heavy workload

Active Publication Date: 2015-06-03
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The operation is cumbersome, the workload is heavy, and the efficiency is low

Method used

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  • Automatic building device of transgenic fish
  • Automatic building device of transgenic fish
  • Automatic building device of transgenic fish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The construction of embodiment 1 transgenic zebrafish

[0041] This example takes the construction of transgenic zebrafish as an example to illustrate the implementation process.

[0042] Since the diameter of the outer membrane of zebrafish embryos is about 1.2mm, embryos at the 1-cell stage are required for injection. When controlling embryos in the system, a transparent tube with an inner diameter of 1.2-1.5mm is used. The transparent tube used in this example is 1.5mm inner diameter of the glass tube.

[0043] Synthesize injection materials using molecular biology methods commonly used to construct transgenic zebrafish, such as Tol2, TALENs / ZFN or CRISPR / Cas9 systems. In this example, the synthesis of CRISPR / Cas9 injection materials refers to the methods in the literatures of Auer et al. (Auer et al., 2014) and Platt et al. (Platt et al., 2014). The mRNAs of Cas9 and sgRAN and the template plasmid containing GPF were synthesized as materials for microinjection.

...

Embodiment 2

[0052] Example 2 Drug injection of zebrafish embryo eggs

[0053] The present invention is not limited to the construction of transgenic zebrafish. The present invention combines the sampling part and the injection part, that is, the output of the sampling part and the input of the injection part are connected through a catheter. To realize the automatic injection of drugs or other solution materials in zebrafish embryos. In this example, taking the injection of ethanol as an example, the toxicological test of ethanol on embryonic development is carried out with zebrafish embryo eggs.

[0054] The embryonic eggs of zebrafish were collected, and the embryonic eggs and the embryonic egg culture solution were poured into the sample tube together. The operation process of the embryonic eggs in the sampling part was the same as that in Example 1. Embryos enter the injection part under the push of a micro-injector, and the operation process for the embryos to enter the injection pa...

Embodiment 3

[0057] The automatic sorting of embodiment 3 zebrafish larvae

[0058] The present invention is not limited to the construction of transgenic zebrafish, and the present invention can realize high-throughput automatic sorting of juvenile fish by combining the sampling part and the sorting part. That is: connect the output of the sampling part with the input of the sorting part.

[0059] This example takes the sorting of albino zebrafish larvae from mixed wild-type and albino zebrafish as an example, and implements the sorting of two kinds of zebrafish larvae. The specific implementation is as follows:

[0060] Put the mixed wild-type zebrafish and albino zebrafish into the sample collection tube. The operation process of the juveniles in the sampling part is the same as that in Example 1. After the juveniles enter the sorting part, locate the juveniles in the first The center of the field of view of the second camera collects images and analyzes the brightness value of the juv...

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Abstract

The invention relates to a device for automatically micro-injecting fish embryonated eggs and automatically sorting juvenile fishes. The device comprises a sampling part and an optionally-injecting part and / or a sorting part, so that the transgenic fish can be effectively built, automatic injection of the fish embryonated eggs is realized, and the automatic sorting of the juvenile fishes is realized.

Description

technical field [0001] The invention relates to the technical field of animal transgenesis, in particular to an automatic construction device and method for transgenic animals, especially transgenic model animals, especially transgenic fish. Background technique [0002] Because model animals such as zebrafish and medaka have the advantages of fast development, strong reproductive ability, in vitro fertilization, in vitro development, transparent body, and small size, they have been widely used in various research and application fields of life medicine. Since the size of zebrafish embryos and hatched juveniles is about 1mm, the construction of transgenic fish needs to rely on manual microinjection of embryos and eggs, and sort the hatched juveniles one by one under a microscope. The operation is cumbersome, the workload is heavy, and the efficiency is low. Contents of the invention [0003] In order to overcome the above-mentioned defects of conventional microinjection, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00
CPCA61M5/31531C12N15/8207
Inventor 黄玉斌胡兵
Owner UNIV OF SCI & TECH OF CHINA
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