CN100363488C - Method for hybridization and fusion of organism germ cell and device thereof - Google Patents
Method for hybridization and fusion of organism germ cell and device thereof Download PDFInfo
- Publication number
- CN100363488C CN100363488C CNB200310106484XA CN200310106484A CN100363488C CN 100363488 C CN100363488 C CN 100363488C CN B200310106484X A CNB200310106484X A CN B200310106484XA CN 200310106484 A CN200310106484 A CN 200310106484A CN 100363488 C CN100363488 C CN 100363488C
- Authority
- CN
- China
- Prior art keywords
- sexual cell
- laser
- biological
- hybridization
- generative cells
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/02—Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Sustainable Development (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Electromagnetism (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention discloses a method and a device for the hybridization and the infusion of biologic generative cells. The method comprises irradiation to generative cells by a laser and artificial fertilization or pollination; particularly, a cold laser of which the energy density is from 0.01 to 2.0 joules/square centimeter and the wave length is from 146 to 532 nanometers as a light source to irradiate the generative cells for 20 femtoseconds to 300 microseconds. The device comprises a laser (21) and the generative cells (27); particularly, an output light path of the laser (21) is provided with a lens (23), a filter (24), a narrow-band reflecting mirror (25) and the generative cells (27), wherein the generative cells (27) are placed in a vessel (28); the narrow-band reflecting mirror (25) is positioned on the light path of a microscope (26). The device can modify an identifying immune system of pellicles of the generative cells or weaken or disturb the identifying immunological functions of the generative cells to be favorable for the hybridization and the fusion of the generative cells in different genera, namely artificial fertilization or pollination to make the generative cells to be hybridized and fused to generate a new biologic variety. The present invention can be widely used for creating new biologic varieties.
Description
Technical field the present invention relates to a kind of method and device of biological hybridization, especially method and the device that impels biological sexual cell hybridization to merge with laser.
Background technology today, science and technology has obtained the development of advancing by leaps and bounds.Yet, people cultivate the research in field in biological new variety, the particularly interbiotic outbreeding research of different genera, but still difficult, this is because biological sexual cell pellicle particularly, in the evolutionary process in millions upon millions of years, possessed intensive identification immunologic function, the gamogenesis cell only between identical kind, under the condition of mutual concession, could merge fertilization.Plant particularly, in process by medium pollination, without any the active selective power, the sheerly passive behavior of being pollinated, fertilization process is finished by the identification immunity system of sexual cell self fully.Therefore, the structure of its sexual cell, fertilization process are all very complicated.Incompatibility in incompatibility and the kind between the kind of biological self reproducing helps safeguarding that the pure of species with stable, multiplies from generation to generation, certainly, also just cultivates biological new variety to artificial hybridization and has caused very big difficulty.People are in order to obtain biological new variety, attempted the whole bag of tricks, the one, the embryo of biology is handled, attempt to make biological gene to produce variation and obtain new variety, as space breeding, plasma body breeding with laser radiation seed etc., its majority is the improvement that belongs to biological variety, so do not create biological new variety; The 2nd, the sexual cell of biology is handled, be intended to directly produce biological new variety, as artificial hybridization fertilization, somatocyte fusion, electricimpulse mediation hybridization etc., do not obtain substantial breakthrough yet.And for example, in the Russian invention patent disclosure specification sheets RU-2035858 that announces May 27 nineteen ninety-five, disclosed a kind of method with ruddiness or infrared band laser bundle irradiation milt daughter cell, but its purpose only is in order to improve the breeding vigor of sexual cell, this method adopts laser beam that the sexual cell of biology is reached radiation treatment more than 10 seconds, the result makes the rate of fertilization of sashimi (raw fish) cell colonization improve 25%, embryo survival has improved 12%, and the speed of growth of fry is accelerated.As seen, this kind method also only is a reproduction activity of having improved the milt daughter cell, has improved the breeding vigor of milt daughter cell, also can not directly not nurture biological new variety.
The summary of the invention the technical problem to be solved in the present invention is for overcoming weak point of the prior art, but a kind of Artificial Control, practicality easy and simple to handle are provided, and equipment is simple, the method and the device that impel the biological sexual cell hybridization of different genera to merge.
The method that biological sexual cell hybridization is merged comprises uses the laser radiation sexual cell, and artificial insemination or pollination, be that 0.01～2.0 Jiao Er/square centimeter, wavelength are that the cold laser of 146～532 nanometers is as light source particularly with energy density, irradiation sexual cell 20 femtoseconds～300 microseconds, wherein, said sexual cell is biological sexual cell, but except people's the sexual cell.
As the further improvement of biological sexual cell hybridization fusion method, described Wavelength of Laser is 193 nanometers or 308 nanometers or 355 nanometers or 532 nanometers; Described sexual cell is female or male; Described sexual cell for exsomatize or at body; Described sexual cell be natural mode or handle through artificial.
The device of implementing biological sexual cell hybridization fusion method comprises laser apparatus, sexual cell, particularly be equipped with lens, spectral filter, narrowband reflection mirror and sexual cell on the output light path of said laser apparatus, said sexual cell places vessel, and said narrowband reflection mirror is positioned on the microscopical light path.
As the further improvement of the device of implementing biological sexual cell hybridization fusion method, described lens are condenser lens; Be equipped with speculum between described laser apparatus and lens; The other supplementary lighting light source that is equipped with of described sexual cell; Described laser apparatus is solid statelaser or gas laser.
Beneficial effect with respect to prior art is, one, because the optical maser wavelength of selecting for use is identical with the resonance absorption frequency range of biological sexual cell pellicle, pellicle has produced the intensive absorption to laser energy, thereby make the pellicle part be decomposed ablation by laser in moment, caused the identification immunity system modification of pellicle, or weaken or disturbed the identification immunologic function of sexual cell, then merge for the hybridization of carrying out between the different genera sexual cell, i.e. artificial insemination or pollination are merged its hybridization possibility are provided; Its two, the sexual cell of irradiated biology be need not to carry out particular processing, make the operating process of laser radiation simple and easy to do; Its three, the simple structure of irradiating unit, use cost is low, is convenient to penetration and promotion.
Description of drawings is described in further detail optimal way of the present invention below in conjunction with accompanying drawing.
Fig. 1 decomposes ablation sexual cell pellicle, makes the synoptic diagram of biological new variety method with laser for the present invention, and wherein, Fig. 1 a be the sexual cell of biology, Fig. 1 b be by the laser ablation sexual cell among Fig. 1 a of part pellicle;
Fig. 2 is a kind of basic structure synoptic diagram of apparatus of the present invention;
Fig. 3 is that laser decomposes the synoptic diagram of ablation animal crucian ovum, and wherein, Fig. 3 a is a primary crucian ovum, Fig. 3 b be by the laser ablation crucian ovum among Fig. 3 a of part pellicle;
Fig. 4 is that laser decomposes the synoptic diagram of ablation animal white mouse sexual cell, and wherein, Fig. 4 a is a white mouse primary spermoblast, Fig. 4 b be by the laser ablation Fig. 4 a spermoblast of part pellicle;
Fig. 5 decomposes ablation plant white trumpet lily at body style synoptic diagram for laser, and wherein, Fig. 5 a is the original style gynoecium of plant white trumpet lily, Fig. 5 b the has been ablation head of Fig. 5 a style gynoecium of part pellicle.
Embodiment is referring to Fig. 1 and Fig. 2, and sexual cell 27 is wrapped in tenuigenin 2 successively outward by nucleus 3 and pellicle 1 constitutes; At occurring in nature, sexual cell 27 as experience are with the different in nature sexual cell 4 of kind, then different in nature sexual cell 4 can waltz through pellicle 1 and tenuigenin 2, to merge fertilization mutually with nucleus 3, as meet with the different in nature animal sexual cell 5 or the plant reproductive cell 6 of different genera, then pellicle 1 and tenuigenin 2 shelter from it, hinder the fusion fertilization of the different in nature animal sexual cell 5 of but different genera or plant reproductive cell 6 and nucleus 3.Be disposed with speculum 22, condenser lens 23, spectral filter 24 on the light path of laser apparatus 21 and be positioned at narrowband reflection mirror 25 on microscope 26 light paths, and the sexual cell 27 that places vessel 28; Wherein, laser apparatus 21 can be solid statelaser or gas laser, the sexual cell 27 other supplementary lighting light sources 29 that are equipped with.During use, the vessel 28 that will be loaded with biological sexual cell 27 earlier place on the focal plane of laser output; Back adjusting laser 21 output laser; Speculum 22 in the re-adjustment laser optical path, make laser beam pass through condenser lens 23, spectral filter 24 after, on narrowband reflection mirror 25, reflect and turn to, shine on the sexual cell 27; Afterwards, transfer microscope 26 to make it to overlap, and can monitor irradiated sexual cell 27 with the direction of laser beam; Then, adjusting laser 21 output optical pulses, when pellicle 1 part of sexual cell 27 is decomposed ablation by laser, when tenuigenin 2 parts are exposed, inject the different in nature animal sexual cell 5 or the plant reproductive cell 6 of different genera biology, as Fig. 1 b, hybridization such as the artificial insemination of enforcement sexual cell 27 or pollination are merged; At last, in microscope 26, monitor the hybridization syncretizing effect of sexual cell 27.
Embodiment 1
Referring to Fig. 2 and Fig. 3, it is the gas laser of 308 nanometers that laser apparatus 21 is selected output wavelength for use, and sexual cell 27 is the stripped ovum 3a of animal crucian, and it is wrapped in tenuigenin 32 successively outward by nucleus 33 and pellicle 31 constitutes.The laser beam of laser apparatus 21 outputs is after speculum 22 is adjusted, shine on the crucian ovum 3a in the vessel 28 by condenser lens 23, spectral filter 24, narrowband reflection mirror 25 again, irradiation energy density is 1.2 Jiao Er/square centimeters, each 14 nanoseconds of irradiation duration, every irradiation is the thick about 0.4 μ m of back ablation pellicle once, and concurrent irradiation 7 times is observed crucian ovum 3a in XS-B1 type microscope 26, its tenuigenin 32 parts are exposed, shown in Fig. 3 b.At this moment, inject carp spermoblast 35, hybridize fusion.In microscope 26, can be observed the fusion of cell.Adjust color, light intensity and the irradiating angle of supplementary lighting light source 29, can obtain the optimal viewing effect.
Referring to Fig. 2 and Fig. 4, it is the gas laser of 193 nanometers that laser apparatus 21 is selected output wavelength for use, and sexual cell 27 is the stripped spermoblast 4a of animal white mouse, and it is made of afterbody (oversize because of it, as not draw among the figure), cell paste 42 and cell head 41.The laser beam of laser apparatus 21 outputs is after speculum 22 is adjusted, shine on the cell head 41 of white mouse spermoblast 4a by condensing lens 23, spectral filter 24, narrowband reflection mirror 5 again, irradiation energy density is 0.8/ square centimeter, 12 nanoseconds of irradiation duration, shines 3 times; The part pellicle of showed cell 41 is by laser about 1 μ m that decomposed ablation, shown in Fig. 4 b in 640 times biomicroscopes 26.The ovum of this cell 4b and rabbit is hybridized, can in microscope 26, observe bicelluar fusion.
Referring to Fig. 2 and Fig. 5, it is the solid statelaser of 532 nanometers that laser apparatus 21 is selected output wavelength for use, sexual cell 27 be the plant white trumpet lily at body style gynoecium 5a, it gynoecium pellicle 52 by lily style gynoecium 51 external parcel and constitutes.Laser beam after speculum 22 is adjusted, again by condensing lens 23, spectral filter 24, narrowband reflection mirror 25 be radiated at white trumpet lily on body style gynoecium 5a, irradiation energy density is 0.07 Jiao Er/square centimeter, irradiation duration 30 psecs are shone 2 times; The part gynoecium pellicle 52 that can be observed lily style gynoecium 5a in microscope 26 is decomposed ablation by laser, shown in Fig. 5 b.Gather chrysanthemum pollen 53, tiger lily pollen 54 and gladiolus pollen 55, respectively white trumpet lily carried out the artificial pollination experiment at body style gynoecium 5b.Through tissue slice, in microscope 26, observe three kinds of pollen and all grown pollen tube.
Obviously, those skilled in the art's method that can merge biological sexual cell hybridization of the present invention and device carry out various changes and modification and do not break away from the spirit and scope of the present invention.Like this, if of the present invention these are revised and modification belongs within the scope of claim of the present invention and equivalent technologies thereof, then the present invention also is intended to comprise these changes and modification interior.
Claims (10)
1. a biological sexual cell is hybridized the method that merges, comprise and use the laser radiation sexual cell, and artificial insemination or pollination, it is characterized in that with energy density being that 0.01～2.0 Jiao Er/square centimeter, wavelength are that the cold laser of 146～532 nanometers is as light source, irradiation sexual cell 20 femtoseconds～300 microseconds, wherein, said sexual cell is biological sexual cell, but except people's the sexual cell.
2. the method that biological sexual cell hybridization according to claim 1 is merged is characterized in that Wavelength of Laser is 193 nanometers or 308 nanometers or 355 nanometers or 532 nanometers.
3. the method that biological sexual cell hybridization according to claim 1 is merged is characterized in that sexual cell is female or male.
4. the method that biological sexual cell hybridization according to claim 1 is merged, it is characterized in that sexual cell be exsomatize or at body.
5. the method that biological sexual cell hybridization according to claim 1 is merged, it is characterized in that sexual cell be natural mode or handle through artificial.
6. implement the device that the described biological sexual cell of claim 1 is hybridized fusion method for one kind, comprise laser apparatus (21) and sexual cell (27), it is characterized in that being equipped with lens (23), spectral filter (24), narrowband reflection mirror (25) and sexual cell (27) on the output light path of said laser apparatus (21), said sexual cell (27) places vessel (28), and said narrowband reflection mirror (25) is positioned on the light path of microscope (26).
7. the device of the biological sexual cell hybridization of enforcement according to claim 6 fusion method is characterized in that lens (23) are condenser lens.
8. the device of the biological sexual cell hybridization of enforcement according to claim 6 fusion method is characterized in that being equipped with speculum (22) between laser apparatus (21) and lens (23).
9. the device of the biological sexual cell hybridization of enforcement according to claim 6 fusion method is characterized in that the other supplementary lighting light source (29) that is equipped with of sexual cell (27).
10. the device of the biological sexual cell hybridization of enforcement according to claim 6 fusion method is characterized in that laser apparatus (21) is solid statelaser or gas laser.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200310106484XA CN100363488C (en) | 2003-11-28 | 2003-11-28 | Method for hybridization and fusion of organism germ cell and device thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB200310106484XA CN100363488C (en) | 2003-11-28 | 2003-11-28 | Method for hybridization and fusion of organism germ cell and device thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1621516A CN1621516A (en) | 2005-06-01 |
| CN100363488C true CN100363488C (en) | 2008-01-23 |
Family
ID=34757596
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200310106484XA Expired - Fee Related CN100363488C (en) | 2003-11-28 | 2003-11-28 | Method for hybridization and fusion of organism germ cell and device thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100363488C (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9249385B1 (en) | 2014-12-18 | 2016-02-02 | City University Of Hong Kong | System and method for fusing cells |
| CN109576152A (en) * | 2019-01-17 | 2019-04-05 | 重庆大学 | A kind of composite pulse cell electricity melts instrument and control method |
-
2003
- 2003-11-28 CN CNB200310106484XA patent/CN100363488C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN1621516A (en) | 2005-06-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Berns et al. | Laser scissors and tweezers | |
| Berns et al. | Laser microbeam as a tool in cell biology | |
| JPS6083584A (en) | Method for ingression of substance into live cell | |
| Hosokawa et al. | Nondestructive isolation of single cultured animal cells by femtosecond laser-induced shockwave | |
| Fajardo et al. | Control of a specific motor program by a small brain area in zebrafish | |
| Macintyre et al. | The responses in culture of human tumour astrocytes and neuroblasts to N 6, O 2′-dibutyryl adenosine 3′, 5′-monophosphoric acid | |
| US20110092966A1 (en) | Ultrashort pulse laser applications | |
| CN1894004A (en) | Near infrared microbial elimination laser system | |
| Karu | Lasers in infertility treatment: irradiation of oocytes and spermatozoa | |
| Sinha et al. | High-speed laser microsurgery of alert fruit flies for fluorescence imaging of neural activity | |
| CN100363488C (en) | Method for hybridization and fusion of organism germ cell and device thereof | |
| Hoffmann | Laser microbeams for the manipulation of plant cells and subcellular structures | |
| Berns | Laser scissors and tweezers to study chromosomes: a review | |
| Schlenk et al. | Laser assisted fixation of ear prostheses after stapedectomy | |
| Neev et al. | Opening of the mouse zona pellucida by laser without a micromanipulator | |
| JP2003530148A (en) | Improved laser comb design / function | |
| Obruca et al. | Ultrastructural observations in human oocytes and preimplantation embryos after zona opening using an erbium-yttrium-aluminium-garnet (Er: YAG) laser. | |
| CN1201653C (en) | Induction method of gynogenesis of multicalor abalone | |
| Yew et al. | Effect of low dose laser on the chorioallantoic culture of retinal pigment cells | |
| Clayton Jr et al. | Individual and population growth in the asexually reproducing anemone Aiptasia pallida Verrill | |
| Whipple et al. | Electron microscopy of CO2‐laser‐induced effects in human fibrocartilage | |
| Berns et al. | Laser applications in biomedicine. Part I: biophysics, cell biology, and biostimulation | |
| JPS6258704B2 (en) | ||
| Tadir | Ten years of laser-assisted gametes and embryo manipulations | |
| Berns | Microbeams |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20080123 Termination date: 20091228 |