CN1428419A - Method for preparing fertilized ovum chromosome - Google Patents
Method for preparing fertilized ovum chromosome Download PDFInfo
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- CN1428419A CN1428419A CN 02110042 CN02110042A CN1428419A CN 1428419 A CN1428419 A CN 1428419A CN 02110042 CN02110042 CN 02110042 CN 02110042 A CN02110042 A CN 02110042A CN 1428419 A CN1428419 A CN 1428419A
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- slide glass
- stationary liquid
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Abstract
The method for preparing fertilized egg chromosome is characterized by adopting the following steps: injecting pregnant mare serum chorionic gonadotropin into mouse abdominal cavity, killing mouse, separating out uterine tube, using culture fluid to flush out fertilizer egg and culturing until the male and female pronuclei are completely eliminated, using pancreatin to make digestion, using sodium citrate solution to make hypotonic, using fixing liquid to make fixation and using Giemsa to stain. The fertilized egg chromosome prepared by said invention can be used for analyzing damage of harmful chemical substance to filial generation. When it is prepared, in a fertilized egg cell the chromosomes of spermand ovum which are completely different in form and are easy to analyze can be clearly showed, and the metakinesis phase is up to 80%.
Description
The present invention relates to a kind of karyomit(e), particularly relate to a kind of method for preparing fertilized ovum chromosome.
Detrimental substance for example lead, mercury, sulfurous gas etc. more and more is subjected to everybody attention to the influence of filial generation.At present, the chromosomal method of a kind of preparation peripheral blood lymphocyte is arranged, the karyomit(e) with this method preparation can be used for the injury of analytical chemistry material to human body, but can not analyze the influence of chemical substance to filial generation.
The purpose of this invention is to provide a kind of method for preparing fertilized ovum chromosome, the fertilized ovum chromosome of preparing with it can remedy the above-mentioned deficiency of prior art.
A kind of method for preparing fertilized ovum chromosome, it is characterized in that to being estrous female or male mice abdominal injection pregnant mare serum sexual cycle, abdominal injection chorionic-gonadotropin hormone again, put to death mouse then, isolate uterine tube, go out zygote with the F10 nutrient solution that contains calf serum, put into incubator immediately, cultivate till the female-male pronucleus disappearance, use trysinization, it is hypotonic to put into liquor sodii citratis, and I fixes with stationary liquid, ovum is chosen on the slide glass, and drip 1 stationary liquid II, and again slide glass is placed among the stationary liquid II and continues fixingly, and then slide glass is moved on to stationary liquid III fix, take out the warm and humid air blow drying of slide glass, slide glass is directly used Giemsa staining.
The fertilized ovum chromosome of the present invention's preparation can be used for analyzing the injury of deleterious chemical substance to filial generation, usage is easy, price is low, can clearly show complete difference, the sperm of being convenient to analyze and the karyomit(e) of ovum on the form during preparation in a fertilized egg cell, metacinesis is up to 80%.
Below by embodiment the present invention is described.
During the preparation fertilized ovum chromosome, earlier to being sexual cycle that the pregnant mare serum of estrous female mice abdominal injection 5 international unit is to bring out superovulation, pneumoretroperitoneum was injected 5 international unit chorionic-gonadotropin hormones (HCG) in 48 hours, 27-28 hour execution mouse after HCG injection, isolate uterine tube, go out zygote with the F10 nutrient solution that contains 30% calf serum (wherein contain Omaine, concentration is 0.05 μ g/ml), put into air immediately and include 5%CO
2Incubator in, till cultivating female-male pronucleus under 37 ℃ and disappearing (about usually 2-3 hour), with 0.5 milliliter of 1% pancreatin 37 ℃ of digestion 5-8 minute, put into 1% liquor sodii citratis that contains 40% calf serum hypotonic 45 minutes, with stationary liquid I (methyl alcohol: Glacial acetic acid: water (volume ratio)=5: 1: 4) fix 5 minutes, (8) choose on the slide glass with ovum, drip 1 stationary liquid II (methyl alcohol: Glacial acetic acid (volume ratio)=3: 1) slide glass is placed among the stationary liquid II continues to fix 5 minutes more immediately thereon, and then slide glass moved on to (methyl alcohol: Glacial acetic acid: fix 1 minute water (volume ratio)=3: 3: 1) among the stationary liquid III, take out the warm and humid air blow drying of slide glass, slide glass directly with 10% Jim Sa (Giemsa) dyeing 18 minutes, promptly gets fertilized ovum chromosome of the present invention.
Pass through of the harm of research chemical substance with the fertilized ovum chromosome that makes to mouse propagation cell, in order to observe influence to the offspring, its method is easy, economical, all has actual application value widely at aspects such as toxicity, pharmacology, ionizing rays, heredity, aristogenesis.
Be the 4.5-5.5 international unit among the present invention consumption male or female mice abdominal injection pregnant mare serum and chorionic-gonadotropin hormone, contain Omaine in the described nutrient solution, its concentration is 0.045-0.055 μ g/ml, and described culture temperature is 36.5-37 ℃; The temperature of digestion is 30-37 ℃.
Claims (4)
1, a kind of method for preparing fertilized ovum chromosome, it is characterized in that to being estrous female or male mice abdominal injection pregnant mare serum sexual cycle, the abdominal injection chorionic-gonadotropin hormone, put to death mouse, isolate uterine tube, go out zygote with the F10 nutrient solution that contains calf serum, put into incubator immediately, cultivate till the female-male pronucleus disappearance, use trysinization, it is hypotonic to put into liquor sodii citratis, and I fixes with stationary liquid, ovum is chosen on the slide glass, drip 1 stationary liquid II immediately thereon, again slide glass is placed among the stationary liquid II and continues fixingly, and then slide glass is moved on to stationary liquid III fix, take out the warm and humid air blow drying of slide glass, slide glass is directly used Giemsa staining.
2, the method for claim 1 is characterized in that describedly being the 4.5-5.5 international unit for consumption male or female mice abdominal injection pregnant mare serum and chorionic-gonadotropin hormone.
3, the method for claim 1 is characterized in that containing Omaine in the described nutrient solution, and concentration is 0.045-0.055 μ g/ml.
4, the method for claim 1 is characterized in that described culture temperature is 36.5-37 ℃; The temperature of digestion is 30-37 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02110042 CN1428419A (en) | 2002-03-01 | 2002-03-01 | Method for preparing fertilized ovum chromosome |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02110042 CN1428419A (en) | 2002-03-01 | 2002-03-01 | Method for preparing fertilized ovum chromosome |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1428419A true CN1428419A (en) | 2003-07-09 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 02110042 Pending CN1428419A (en) | 2002-03-01 | 2002-03-01 | Method for preparing fertilized ovum chromosome |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102465168A (en) * | 2010-11-18 | 2012-05-23 | 天津市新药安全评价研究中心 | Method for preparing metaphase specimen of mammalian cell chromosome |
| CN112652222A (en) * | 2020-04-30 | 2021-04-13 | 华南农业大学 | Intra-ovarian fertilization process of female silkworm moths and model display method of ovaries |
-
2002
- 2002-03-01 CN CN 02110042 patent/CN1428419A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102465168A (en) * | 2010-11-18 | 2012-05-23 | 天津市新药安全评价研究中心 | Method for preparing metaphase specimen of mammalian cell chromosome |
| CN112652222A (en) * | 2020-04-30 | 2021-04-13 | 华南农业大学 | Intra-ovarian fertilization process of female silkworm moths and model display method of ovaries |
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| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |