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Chromosome slide preparing method

A technology of chromosome production and chromosome, which is applied in the fields of genetics and chromosome analysis, and can solve the problems of high surface tension of glass slides, failure to form water film, insufficient dispersion of chromosome division phase, etc.

Active Publication Date: 2013-09-18
JILIN TUO HUA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that the humidity of the air is relatively high, and the humidity of the air must exceed 60%; and the operation of the drop slide is required quickly, otherwise the slide cannot quickly form a water film in the air
Due to the high surface tension of the glass slide, the cell suspension encountered in liquid dispersion is large, and the resulting chromosomal division phase is often agglomerated or superimposed due to insufficient dispersion, and cannot or rarely be fully dispersed. Chromosomes with clear bands and regular shapes cannot be used for chromosome counting and karyotype analysis

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0034] Example 1: Stimulated activation of blood lymphocytes

[0035] In a T12.5 culture flask, add 8ml of RPMI1640 medium containing 20% ​​calf serum, and add phytohemagglutinin (PHA) with a final concentration of 5mg / ml, then slowly add 1ml of heparin anticoagulated whole blood, 37 °C, 5% CO 2 Cultured for 72 hours; phytohemagglutinin can stimulate in G 0 Phase blood lymphocytes enter G 1 Phase, that is, the cells enter the cell division cycle from the quiescent phase, and the number of lymphocytes stimulated to enter the metaphase of cell division is the largest at 68-72 hours. Shake the culture bottle every 24 hours to mix the blood and avoid clots. To prevent cell death due to aggregation. After culturing for 71 hours, 100 μl of 50 μg / ml colchicine was added, and the culture was continued for 1 hour. The purpose of adding colchicine is to make the lymphocytes in the cell division cycle stagnate in the middle phase of cell division to the greatest extent.

Embodiment 2

[0036] Example 2: Collection and fixation of lymphocytes

[0037] Place the cultured blood lymphocyte suspension in a 10ml centrifuge tube, centrifuge at 2000rpm for 8 minutes, discard the supernatant, keep the precipitated part, and be careful not to damage the buffy coat. Add 8ml of hypotonic solution, mix well, and bathe in water at 37°C for 30 minutes. Add 1ml of fixative, mix thoroughly, centrifuge at 2000rpm for 8 minutes, and discard the supernatant. Add 8ml of fixative, mix thoroughly, centrifuge at 2000rpm for 8 minutes, discard the supernatant, repeat 2 times. Depending on the amount of cell sedimentation, add 0.5-1ml of fixative solution and mix well.

Embodiment 3

[0038] Example 3: Treatment of slides (traditional method)

[0039] 1) Depending on the initial cleanliness of the slides, the slides can be washed with tap water as needed, and then rinsed with distilled water several times, such as 1-3 times.

[0040] 2) Air dry.

[0041] 3) Freeze the dried slides in -20°C refrigerator for 1 hour in advance, and take out the pre-cooled slides directly for dropping slides.

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Abstract

The invention relates to a chromosome slide preparing method which comprises the following steps of: providing a chromosome sample, putting a slide into water and keeping for more than 2 hours at 2-8 DEG C, carrying out pretreatment on the slide, fixing the chromosome sample, and dripping on the slide. The chromosome obtained by using the improved chromosome slide preparing method is sufficient in dispersion, free of overlapping, free of loss, clear in strip and regular in morphology; and the analysis on chromosome karyotype and the diagnosis on diseases are facilitated.

Description

technical field [0001] The invention belongs to the technical field of genetics. Specifically, the present invention relates to the technical field of chromosome analysis; more specifically, it relates to a method for chromosome preparation. Background technique [0002] Chromosomes are the genetic material in the nucleus of a cell. After the lymphocytes in the blood are cultured under certain conditions, they are harvested, made into slices, digested with trypsin, and stained to show the bands. Chromosomes can be observed under a microscope. Humans have 23 pairs of chromosomes, of which 22 are autosomes and 1 is the sex chromosome that determines sex. Genes on chromosomes that carry genetic information. An increase or decrease in the number of chromosomes, or structural abnormalities such as deletions and translocations of chromosomes are called chromosomal diseases, and corresponding clinical symptoms may appear. Whether the number and structure of chromosomes are norm...

Claims

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Application Information

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IPC IPC(8): G01N1/28
Inventor 聂德志宋广赜黄若洋
Owner JILIN TUO HUA BIOTECH
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