CN101305709B - Application of micrometre flimsy in making the nematode permanent glassslide - Google Patents
Application of micrometre flimsy in making the nematode permanent glassslide Download PDFInfo
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- CN101305709B CN101305709B CN2008100585236A CN200810058523A CN101305709B CN 101305709 B CN101305709 B CN 101305709B CN 2008100585236 A CN2008100585236 A CN 2008100585236A CN 200810058523 A CN200810058523 A CN 200810058523A CN 101305709 B CN101305709 B CN 101305709B
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Abstract
The invention relates to the application of a micron tissue in a permanent nematode glassslide, which belongs to the technology field of specimen preparation. In the invention, according to the size of the specimen target, a micron tissue is selected to replace a wax preparing ring and the a support placed in the conventional permanent glassslide preparation, and is used for the preparation of the permanent nematode glassslide. The complex preparation procedures in the conventional permanent nematode glassslide preparation is simplified, the preparation difficulty is reduced, the glassslide preparation quality and the success ratio are improved, and a novel method with simple operation, low cost and high efficiency is provided for the preparation of permanent nematode glassslide specimen. The application not only can be used for the preparation of permanent nematode glassslide specimen, but also can be used for the preparation of various specimens.
Description
Technical field:
The present invention relates to a kind of micron thin paper in nematode permanent slide Applications in Fabrication, belong to preparation of specimen's technical field
Background technology:
Phytopathy original slide sample is that experiment material phytopathy original slide sample important in plant pathology teaching and the scientific research divides permanent slide samples such as permanent slide and semipermanent slide sample forever to preserve, the permanent use.Semipermanent slide sample storage life is short, about 3 years service lives.About nematode permanent slide preparation of specimen method, in monograph (planting the disease research method) and textbook (guide book of experiment), detailed introduction is arranged all.For a long time, the conventional method of system wax ring and placement supporter is adopted in the making of nematode permanent slide always.But in practical operation, the system wax ring in the conventional method is placed supporter, and three step difficulty of control target field range height is time-consuming again, and often a certain slight error will cause that all that has been achieved is spoiled.Therefore for many years nematode permanent slide sample be made into the problem that many researchers avoid.
By literature search, do not see the open report identical with the present invention
Summary of the invention:
The object of the invention is to overcome the deficiency of existing slide sample preparation method, according to the size of purpose sample, selects the alternative conventional permanent slide wax preparing ring of micron thin paper and places supporter, is used for the making of nematode and other pathogen permanent slides.
Select the micron thin paper to substitute conventional permanent slide wax preparing ring and supporter, the concrete steps that are used to make the nematode permanent slide are as follows:
(1), cleans slide and cover glass;
(2), select the thin paper of 28.5 micron thickness for use, be that 15 millimeters circular card punch cut a micron thin paper sheet with diameter;
(3), the circular card punch with 5 millimeters of diameters punches in (2) thin paper center;
(4), solid paraffin is placed culture dish, be placed on the thermostatic electrothermal plate, under 80 ℃ of-90 ℃ of temperature, be heated to solid paraffin and melt fully, keep temperature, wax liquid is not solidified, do not seethe with excitement;
(5), the round scraps of paper that will have openning hole are dipped in the wax liquid of culture dish, fully soak into, side by side its surperficial minute bubbles;
(6), the round scraps of paper that will fully soak into wax liquid with pincet take out, and are put in the slide center position that cleans up rapidly;
(7), the slide that is placed with the waxdip circle scraps of paper on it is placed on the thermostatic electrothermal plate, under 80 ℃ of-90 ℃ of temperature, heat, closely contact fully with slide to the circle scraps of paper, take off after not having bubble between the two, be put on the horizontal stand room temperature cooling;
(8), in the middle of the waxdip circle scraps of paper, splash into 2 milliliters of floating supporting agent glycerine in the roundlet, make its drop circular as far as possible;
(9), the nematode that will dewater is chosen in (8) circular drop arrangement position;
(10), cover glass that (1) is cleaned up is placed on directly over (8) waxdip circle scraps of paper, integral body is put in 80 ℃ of-90 ℃ of heating on the thermostatic electrothermal plate, closely contacts with slide with choosing pin light cap slide, does not extremely have till the bubble between the two;
(11), slide in (10) taken off be put on the horizontal stand room temperature and be cooled to wax liquid and solidify, the sealing cover glass is labelled, and completes.
The present invention has simplified the production process of the conventional nematode permanent slide complexity in making, and has reduced manufacture difficulty, has improved slice-making quality and success rate greatly, provides a kind of easy and simple to handle, cost is low, efficient is high new method for making nematode permanent slide sample.
The present invention not only is used for the making of nematode permanent slide sample, also can be used for the making of multiple sample.
Embodiment:
The present invention prepares according to conventional method.The present invention not only is confined to embodiment and limits, and also can select the thin paper of micron to substitute system wax ring and supporter in the conventional permanent slide, is used for the making of multiple sample.
Embodiment 1:
The making step of pine wood nematode permanent slide sample:
(1), cleans slide and cover glass;
(2), select the thin paper of 28.5 micron thickness for use, be that 15 millimeters circular card punch cut a micron thin paper sheet with diameter;
(3), the circular card punch with 5 millimeters of diameters punches in (2) thin paper center;
(4), solid paraffin is placed culture dish, be placed on the thermostatic electrothermal plate, under 80 ℃ of-90 ℃ of temperature, be heated to solid paraffin and melt fully, keep temperature, wax liquid is not solidified, do not seethe with excitement;
(5), the round scraps of paper that will have openning hole are dipped in the wax liquid of culture dish, fully soak into, side by side its surperficial minute bubbles;
(6), the round scraps of paper that will fully soak into wax liquid with pincet take out, and are put in the slide center position that cleans up rapidly;
(7), the slide that is placed with the waxdip circle scraps of paper on it is placed on the thermostatic electrothermal plate, under 80 ℃ of-90 ℃ of temperature, heat, closely contact fully with slide to the circle scraps of paper, take off after not having bubble between the two, be put on the horizontal stand room temperature cooling;
(8), in the middle of the waxdip circle scraps of paper, splash into 2 milliliters of floating supporting agent glycerine in the roundlet, make its drop circular as far as possible;
(9), the pine wood nematode that will dewater is chosen in (8) circular drop arrangement position;
(10), cover glass that (1) is cleaned up is placed on directly over (8) waxdip circle scraps of paper, integral body is put in 80 ℃ of-90 ℃ of heating on the thermostatic electrothermal plate, closely contacts with slide with choosing pin light cap slide, does not extremely have till the bubble between the two;
(11), slide in (10) taken off be put on the horizontal stand room temperature and be cooled to wax liquid and solidify, the sealing cover glass is labelled, and completes.
Embodiment 2:
The making step of root-knot nematode 2 instar larvae permanent slide samples:
(1), cleans slide and cover glass;
(2), select the thin paper of 28.5 micron thickness for use, be that 15 millimeters circular card punch cut a micron thin paper sheet with diameter;
(3), the circular card punch with 5 millimeters of diameters punches in (2) thin paper center;
(4), solid paraffin is placed culture dish, be placed on the thermostatic electrothermal plate, under 80 ℃ of-90 ℃ of temperature, be heated to solid paraffin and melt fully, keep temperature, wax liquid is not solidified, do not seethe with excitement;
(5), the round scraps of paper that will have openning hole are dipped in the wax liquid of culture dish, fully soak into, side by side its surperficial minute bubbles;
(6), the round scraps of paper that will fully soak into wax liquid with pincet take out, and are put in the slide center position that cleans up rapidly;
(7), the slide that is placed with the waxdip circle scraps of paper on it is placed on the thermostatic electrothermal plate, under 80 ℃ of-90 ℃ of temperature, heat, closely contact fully with slide to the circle scraps of paper, take off after not having bubble between the two, be put on the horizontal stand room temperature cooling;
(8), in the middle of the waxdip circle scraps of paper, splash into 2 milliliters of floating supporting agent glycerine in the roundlet, make its drop circular as far as possible;
(9), the root-knot nematode that will dewater is chosen in (8) circular drop arrangement position;
(10), cover glass that (1) is cleaned up is placed on directly over (8) waxdip circle scraps of paper, integral body is put in 80 ℃ of-90 ℃ of heating on the thermostatic electrothermal plate, closely contacts with slide with choosing pin light cap slide, does not extremely have till the bubble between the two;
(11), slide in (10) taken off be put on the horizontal stand room temperature and be cooled to wax liquid and solidify, the sealing cover glass is labelled, and completes.
Embodiment 3:
The making step of melon and fruit corruption permant preparation of mould sexual generation:
(1), cleans slide and cover glass;
(2), select the thin paper of 50 micron thickness for use, be that 15 millimeters circular card punch cut a micron thin paper sheet with diameter;
(3), the circular card punch with 5 millimeters of diameters punches in (2) thin paper center;
(4), solid paraffin is placed culture dish, be placed on the thermostatic electrothermal plate, under 80 ℃ of-90 ℃ of temperature, be heated to solid paraffin and melt fully, keep temperature, wax liquid is not solidified, do not seethe with excitement;
(5), the round scraps of paper that will have openning hole are dipped in the wax liquid of culture dish, fully soak into, side by side its surperficial minute bubbles;
(6), the round scraps of paper that will fully soak into wax liquid with pincet take out, and are put in the slide center position that cleans up rapidly;
(7), the slide that is placed with the waxdip circle scraps of paper on it is placed on the thermostatic electrothermal plate, under 80 ℃ of-90 ℃ of temperature, heat, closely contact fully with slide to the circle scraps of paper, take off after not having bubble between the two, be put on the horizontal stand room temperature cooling;
(8), in the middle of the waxdip circle scraps of paper, splash into 2 milliliters of floating supporting agent glycerine in the roundlet, make its drop circular as far as possible;
(9), mycelia that will dewater and egg spore move in (8) circular drop arrangement position;
(10), cover glass that (1) is cleaned up is placed on directly over (8) waxdip circle scraps of paper, integral body is put in 80 ℃ of-90 ℃ of heating on the thermostatic electrothermal plate, closely contacts with slide with choosing pin light cap slide, does not extremely have till the bubble between the two;
(11), slide in (10) taken off be put on the horizontal stand room temperature and be cooled to wax liquid and solidify, the sealing cover glass is labelled, and completes.
Claims (1)
1. the micron thin paper is characterized in that the size according to the sample object in nematode permanent slide Applications in Fabrication, selects the alternative conventional permanent slide wax preparing ring of micron thin paper and places supporter, is used for the making of nematode permanent slide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100585236A CN101305709B (en) | 2008-06-13 | 2008-06-13 | Application of micrometre flimsy in making the nematode permanent glassslide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008100585236A CN101305709B (en) | 2008-06-13 | 2008-06-13 | Application of micrometre flimsy in making the nematode permanent glassslide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101305709A CN101305709A (en) | 2008-11-19 |
| CN101305709B true CN101305709B (en) | 2011-04-20 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2008100585236A Expired - Fee Related CN101305709B (en) | 2008-06-13 | 2008-06-13 | Application of micrometre flimsy in making the nematode permanent glassslide |
Country Status (1)
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| CN (1) | CN101305709B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102640742A (en) * | 2012-04-20 | 2012-08-22 | 杭州市农业科学研究院 | Improved manufacturing method for plant wireworm permanent slide |
| CN104374616A (en) * | 2014-10-09 | 2015-02-25 | 宁波检验检疫科学技术研究院 | Manufacturing method of plant pathogenic fungus sealed slide glass specimen |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1335061A (en) * | 2000-07-20 | 2002-02-13 | 广东省昆虫研究所 | Storing method of entomopathogenic nematode |
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2008
- 2008-06-13 CN CN2008100585236A patent/CN101305709B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1335061A (en) * | 2000-07-20 | 2002-02-13 | 广东省昆虫研究所 | Storing method of entomopathogenic nematode |
Non-Patent Citations (1)
| Title |
|---|
| 徐克林.植物线虫的采集、分离和固定.中国森林病虫.1984,(3),43-45. * |
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| CN101305709A (en) | 2008-11-19 |
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Granted publication date: 20110420 Termination date: 20120613 |