Preparation method of planarian chromosome specimen

A chromosome and planarian technology, applied in the field of cytogenetics, can solve problems such as unfavorable research and analysis, difficult sampling and processing, unstable results, etc., and achieve the effect of good cell dispersion, clear background and good effect

Inactive Publication Date: 2016-04-20
ZHOUKOU NORMAL UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] The existing mature animal chromosome specimen preparation methods are mostly suitable for higher vertebrates and humans, and can be prepared after processing by collecting bone marrow or blood; while planarians only have tissue fluid, no spinal cord and blood, so sampling and processing are difficult
Most of the existing technologies use the method of Li Guangpeng ("Journal of Zoology" 1992, 27 (5), 30~31), but the results are unstable and the film production is not clear, which is not conducive to research and analysis, and there is a large room for improvement

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  • Preparation method of planarian chromosome specimen

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Embodiment 1

[0029] The preparation method of the planarian chromosome specimen in this embodiment, the reagents used are as follows: fixative solution I is composed of ethanol and glacial acetic acid in a volume ratio of 4:1, fixative solution II is composed of ethanol and glacial acetic acid in a volume ratio of 2:1, and fixative solution III is composed of ethanol and glacial acetic acid in a volume ratio of 2:1. Composed of ethanol and glacial acetic acid with a volume ratio of 1:4, the Giemsa staining solution is a Giemsa grinding solution containing 10wt% glycerol;

[0030] This embodiment mainly has the following steps:

[0031] (1) Material collection and pretreatment: Select a lively and healthy planarian with a body length of 3 cm, put it in dechlorinated tap water, and starve it for 1 week until no metabolic waste is discharged;

[0032] (2) Regeneration tissue culture: place the planarian treated in step (1) on a glass slide, and when it is fully stretched, cut the planarian in...

Embodiment 2

[0039] The preparation method of the planarian chromosome specimen in this embodiment, the reagents used are as follows: fixative solution I is composed of ethanol and glacial acetic acid in a volume ratio of 4:1, fixative solution II is composed of ethanol and glacial acetic acid in a volume ratio of 2:1, and fixative solution III is composed of ethanol and glacial acetic acid in a volume ratio of 2:1. Composed of ethanol and glacial acetic acid at a volume ratio of 1:4;

[0040] This embodiment mainly has the following steps:

[0041] (1) Material collection and pretreatment: Select a lively and healthy planarian with a body length of 5 cm, put it in dechlorinated tap water, and starve it for 2 weeks until no metabolic waste is discharged;

[0042] (2) Regeneration tissue culture: place the planarian treated in step (1) on a glass slide, and when it is fully stretched, cut the planarian into 5 sections with an ethanol-sterilized blade and place it at a constant temperature o...

Embodiment 3

[0049] The preparation method of the planarian chromosome specimen in this embodiment, the reagents used are as follows: fixative solution I is composed of ethanol and glacial acetic acid in a volume ratio of 4:1, fixative solution II is composed of ethanol and glacial acetic acid in a volume ratio of 2:1, and fixative solution III is composed of ethanol and glacial acetic acid in a volume ratio of 2:1. Composed of ethanol and glacial acetic acid with a volume ratio of 1:4, the Giemsa staining solution is a Giemsa grinding solution containing 20wt% glycerol;

[0050] This embodiment mainly has the following steps:

[0051] (1) Material collection and pretreatment: Select a lively and healthy planarian with a body length of 4 cm, put it in dechlorinated tap water, and starve it for 10 days until no metabolic waste is discharged;

[0052] (2) Regenerative tissue culture: place the planarian treated in step (1) on a glass slide, and when it is fully stretched, cut the planarian i...

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Abstract

The invention relates to a method for preparing a planarian chromosome sample. The preparation method of the planarian chromosome specimen of the present invention comprises the following steps: material collection, pretreatment, regeneration tissue culture, tissue crushing, centrifugal suspension, colchicine blocking, KC1 hypotonic treatment, centrifugal suspension dripping, dyeing and drying and sealing piece. The preparation method of the invention fully fixes the regenerated tissue and the metaphase cells, has a clear chromosome preparation background, well-dispersed cells, and many metaphase division phases, which is beneficial to karyotype analysis.

Description

technical field [0001] The invention belongs to the field of cytogenetics, and in particular relates to a method for preparing a planarian chromosome sample. Background technique [0002] Planarians are widely distributed in my country. In the history of animal evolution, planarians are the first group with bilateral symmetry and three germ layers. Their evolutionary status is very important, and they are good materials for the research of developmental biology, cytogenetics and regenerative biology. Chromosome preparation of planarian is the basis of its genetics research, as well as the basis of cell and molecular biology research. [0003] The existing mature methods for preparing animal chromosome specimens are mostly suitable for higher vertebrates and humans, and can be prepared after processing by collecting bone marrow or blood; while planarians only have interstitial fluid, no spinal cord and blood, so sampling and processing are difficult. Li Guangpeng (Journal of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/30
Inventor 马克世盛东峰武安泉王永立杨同文陈龙
Owner ZHOUKOU NORMAL UNIV
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