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Chromosome specimen preparation liquid for laser microdissection and application thereof

A technology of laser microdissection and specimen preparation, which is applied in the field of laser microdissection chromosome specimen preparation solution, which can solve the problems of few research reports and difficulty of chromosome division phase, etc., and achieve the effect of convenient use, simple and practical preparation method, and easy operation

Inactive Publication Date: 2013-01-09
OCEAN UNIV OF CHINA
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Problems solved by technology

But so far, the laser microdissection system is mostly used for the cutting and separation of various cells and tissues, but there are only few research reports on the microdissection of chromosomes.
The reason is that most of the laser microdissection systems use a special surface-coated glass slide when performing microdissection, and on this glass slide are prepared a good shape, a large number and can be used for cutting. Chromosome division phase has great difficulty

Method used

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  • Chromosome specimen preparation liquid for laser microdissection and application thereof
  • Chromosome specimen preparation liquid for laser microdissection and application thereof

Examples

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Embodiment Construction

[0019] The chromosome sample preparation solution and preparation method of the present invention will be described in detail below in conjunction with the cutting of the W chromosome of the female semi-smooth tongue sole:

[0020] (1) Chromosome preparation

[0021] ①. The head kidney tissue of tongue sole was taken to make cell suspension by physical crushing method, and treated with 0.005% colchicine for 1.5 hours;

[0022] ② After the cells were collected by centrifugation, they were hypotonically treated with 0.075M KCL hypotonic solution for 30 minutes;

[0023] ③. Collect by centrifugation again, fix with Carnot’s fixative (anhydrous ethanol: glacial acetic acid = 3:1) 3 times, 20 minutes each time, store at -20°C after the last fixation;

[0024] ④. Before making slices, centrifuge the cells and resuspend them in the laser microdissection chromosome sample preparation solution. The cell concentration is about 1×10 6 Left and right, make working solution of chromosome...

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Abstract

The invention relates to a chromosome specimen preparation liquid for laser microdissection and application thereof. The specimen preparation liquid comprises methanol, glacial acetic acid and chloroform of which the volume ratio is (5-10):(1-3):(1-3), preferably 8:1:1. The chromosome specimen preparation liquid provided by the invention is used for preparing a chromosome specimen for laser microdissection. In the chromosome specimen for laser microdissection, which is prepared by the chromosome specimen preparation liquid, the chromosomes are well dispersed, thereby facilitating the subsequent microdissection operation. Meanwhile, the preparation method provided by the invention is simple and practical, is easy to operate, and is convenient for common technical personnel to use.

Description

technical field [0001] The invention belongs to the technical field of chromosome laser microdissection, and in particular relates to a laser microdissection chromosome specimen preparation solution and application thereof. Background technique [0002] Chromosome cutting methods can be divided into two types, namely microscopic glass needle separation method and laser cutting method. Microscopic glass needle separation method uses a micromanipulator to cut and separate the complete target chromosome or target banded segment with a glass tip diameter of about 2 microns under a microscope. This technique has a wide range of applications, but requires a micromanipulator with good adjustment performance and large enough magnification, and requires the experimenter to have very high operating skills. The laser cutting method uses a laser micro-beam with a wavelength of 340-355nm with a low thermal effect, and controls its trajectory through a computer to cut the target chromoso...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 王旭波张全启齐洁王志刚于海洋
Owner OCEAN UNIV OF CHINA
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