CN105181406A - Method for making paraffin section of elytrum of insect - Google Patents
Method for making paraffin section of elytrum of insect Download PDFInfo
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- CN105181406A CN105181406A CN201510505600.8A CN201510505600A CN105181406A CN 105181406 A CN105181406 A CN 105181406A CN 201510505600 A CN201510505600 A CN 201510505600A CN 105181406 A CN105181406 A CN 105181406A
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Abstract
The invention provides a method for making a paraffin section of the elytrum of an insect. The method comprises the following steps: 1, distinguishing female and male coleoptera insects in a pupal stage and taking the elytra of the insects after eclosion for preparation of paraffin sections; 2, fixing the elytra, i.e., cutting the to-be-observed parts of the elytra obtained in the step 1 and fixing the parts with an FAA fixing agent; 3, dehydrating the elytra, i.e., successively dehydrating the fixed elytrum tissue with alcohols with different volume concentrations; 4, transparentizing the elytra, i.e., transparentizing the dehydrated elytra in a solution of anhydrous alcohol and xylene at first and then in xylene; 5, impregnating the elytra with paraffin, i.e., putting the transparentized elytra in a solution of soft paraffin and xylene with a temperature of 55 DEG C for 3 h at first and then in a solution of hard paraffin and xylene with a temperature of 65 DEG C for 3 h, and then carrying out excessive paraffin impregnation at a constant temperature of 65 DEG C for 6 h; 6, carrying out embedding and slicing; 7, carrying out unfolding, pasting and roasting; 8, carrying out paraffin removal and rehydration; and 9, carrying out dyeing and mounting. The elytrum paraffin section prepared by using the method has a high integrity rate and can meet requirements of research on the microstructure and ultrastructure of the elytra.
Description
Technical field
The present invention relates to microscopic tissue sections technical field, be specifically related to a kind of insect elytrum paraffin section method.
Background technology
The fore wing of coleopteron be it is generally acknowledged and all ossify, only for the protection of back and hind wing.Existing to neurocyte in coleopteron wing and neural research extremely weak and ossified owing to elytrum of this situation.But actual conditions make the neurocyte distributed in elytrum be difficult to observe instead of do not have what tissue alive and cell in elytrum.
Yellow meal worm adult has cornified elytrum, larva brown color, eating flour, so cry yellow meal worm, cries again bread worm, and formal name used at school Tenebriomolitor (L.) belongs to coleoptera, intends Paussidae, intends ground beetle and belongs to.Yellow meal worm is that (cultivation of yellow meal worm is based on wheat bran of throwing something and feeding, rice bran for omnivorous insect, suitably add and feed fish meal, beancake powder, coarse colza meal, respectively to plant vegetables leaf etc.), be a kind of large-scale storage pest, be also the valuable source insect of great exploitation potential for its, a kind of especially can at the important Physiology Experiment material of laboratory rearing.
Up-to-date research finds that elytrum surface has in sensation hair, elytrum and has nerve alive, and neurocyte has the function of generations, conduction and global neurological excitement and nerve information, has important effect to the activity of insect.And feel that the research of hair and neurocyte needs to carry out elytrum section the research of anatomical structure; Tracheole systematic research in elytrum also has pending Ultrastructural analysis.Original paraffin section method, because elytrum quality is hard, the effect of paraffin infiltration is very bad, the percentage of damage that elytrum is cut into slices is high, higher than 70%, can not meet the requirement of the inside Microstructure and fabric observing elytrum, this is also main cause weak to the research of insect elytrum at present.
Summary of the invention
Based on above weak point, the object of the present invention is to provide a kind of insect elytrum paraffin section method, for the requirement of the inside Microstructure and fabric of satisfied observation elytrum.
The technology used in the present invention is as follows: a kind of insect elytrum paraffin section method, as follows:
Step (1) is distinguished coleopteron female, male in pupa time, gets its elytrum for paraffin section after emergence in the 36-48 little time;
Step (2) elytrum is fixed: the Partial Shear will observed by the elytrum that step (1) obtains is good, fixes 24 hours with FAA fixing agent;
Step (3) elytrum dewaters: the alcohol of fixing elytrum tissue different volumes concentration is dewatered successively, and dehydration concentration and time are followed successively by: 50%1-2 hour, 70%1-2 hour, 80%30 minutes, 90%30 minutes, absolute alcohol 20 minutes, absolute alcohol 10 minutes;
Step (4) elytrum is transparent: put into by volume by the elytrum of dehydration, anhydrous alcohol: dimethylbenzene=1: transparent 1-2 hour in the liquid of 1, then puts into dimethylbenzene 1 hour;
Step (5) elytrum waxdip: the elytrum of transparent mistake is put into 55 DEG C, melt, by volume, soft wax: dimethylbenzene=1: in 1 liquid 3 hours; Put into again 65 DEG C that melted, by volume, hard paraffin: dimethylbenzene=1: in 1 liquid 3 hours; 65 DEG C of constant temperature are after spending waxdip 6 hours;
Step (6) embedding and section: take out elytrum, cut into slices after coagulation forming in uncle's mouthful carton;
Step (7) is opened up sheet, bonding die and is copied sheet;
Step (8) dewaxing rehydration;
Mounting after step (9) dyeing.
Adopt the insect elytrum paraffin section after method process of the present invention, percentage of damage is very low, and percentage of head rice is high, reaches the percentage of head rice of more than 90%, can meet the requirement of its Microstructure and fabric of research.
Embodiment
The invention will be further described in citing below.
Embodiment 1:
By the yellow meal worm raised, separate female, male pupa in pupa time, more female, male pupa is placed in raising capsule respectively, female after sprouting wings, male imago has just separated (because yellow meal worm adult is difficult to distinguish male and female) naturally.
The elytrum getting the female worm of yellow meal worm (or male worm, the needs by research) of after sprouting wings 36-48 hour puts into FAA immobile liquid.Take out after one hour, put into FAA immobile liquid after the Partial Shear that will observe is good again and continue to fix 23 hours.
The elytrum tissue gradient alcohol dehydration after fixing.The inchoate aspect volume concentrations of dehydrated alcohol is 50%, and in the alcohol of variable concentrations, dewatering time is incomplete same.Be followed successively by: 50%1-2 hour, 70%1-2 hour, 80%30 minutes, 90%30 minutes, absolute alcohol 20 minutes, absolute alcohol 10 minutes.
Yellow meal worm elytrum tissue after dehydration is put into, is the anhydrous alcohol of 1: 1 by volume: transparent 1-2 hour in the liquid of dimethylbenzene, then puts into dimethylbenzene transparent 1 hour.
Low melt point paraffin is placed in capsule, is placed in 55 DEG C of constant temperature ovens and melts, add dimethylbenzene, by volume soft wax: dimethylbenzene=1: 1 in batches.The elytrum of transparent mistake is put into, adds a cover standing 3 hours.Put into 65 DEG C of melted, hard paraffins by volume again: dimethylbenzene=1: 1 liquid 3 hours; Remove the paraffin liquid of 1/2, then add the paraffin liquid of equivalent, 65 DEG C of waxdips 2 hours; Pour out the paraffin liquid of 1/2 again, then add the paraffin liquid of equivalent, waxdip 2 hours; 65 DEG C of paraffin refined wax immersion wax 2 hours.
Cut into slices after coagulation forming in uncle's mouthful carton.
After this exhibition sheet, bonding die and copy sheet, dewaxing rehydration, after dyeing, mounting is all conventional method.
The yellow meal worm elytrum paraffin section of such process, percentage of damage is very low, and percentage of head rice is very high, reaches the percentage of head rice of more than 90%, can meet the requirement of its Microstructure and fabric of research.
Claims (1)
1. an insect elytrum paraffin section method, is characterized in that, method is as follows:
Step (1) is distinguished coleopteron female, male in pupa time, gets its elytrum for paraffin section after emergence in the 36-48 little time;
Step (2) elytrum is fixed: the Partial Shear will observed by the elytrum that step (1) obtains is good, fixes 24 hours with FAA fixing agent;
Step (3) elytrum dewaters: the alcohol of fixing elytrum tissue different volumes concentration is dewatered successively, and dehydration concentration and time are followed successively by: 50%1-2 hour, 70%1-2 hour, 80%30 minutes, 90%30 minutes, absolute alcohol 20 minutes, absolute alcohol 10 minutes;
Step (4) elytrum is transparent: put into by volume by the elytrum of dehydration, anhydrous alcohol: dimethylbenzene=1: transparent 1-2 hour in the liquid of 1, then puts into dimethylbenzene 1 hour;
Step (5) elytrum waxdip: the elytrum of transparent mistake is put into 55 DEG C, melt, by volume, soft wax: dimethylbenzene=1: in 1 liquid 3 hours; Put into again 65 DEG C that melted, by volume, hard paraffin: dimethylbenzene=1: in 1 liquid 3 hours; 65 DEG C of constant temperature are after spending waxdip 6 hours;
Step (6) embedding and section: take out elytrum, cut into slices after coagulation forming in uncle's mouthful carton;
Step (7) is opened up sheet, bonding die and is copied sheet;
Step (8) dewaxing rehydration;
Mounting after step (9) dyeing.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510505600.8A CN105181406B (en) | 2015-08-14 | 2015-08-14 | A kind of insect elytrum paraffin section method |
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| CN201510505600.8A CN105181406B (en) | 2015-08-14 | 2015-08-14 | A kind of insect elytrum paraffin section method |
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| CN105181406A true CN105181406A (en) | 2015-12-23 |
| CN105181406B CN105181406B (en) | 2018-04-03 |
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| CN201510505600.8A Expired - Fee Related CN105181406B (en) | 2015-08-14 | 2015-08-14 | A kind of insect elytrum paraffin section method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105928752A (en) * | 2016-04-13 | 2016-09-07 | 沈阳大学 | Method for staining paraffin section of adult insect |
| CN106568632A (en) * | 2016-11-08 | 2017-04-19 | 沈阳农业大学 | Method of converting paraffin slice sample into scanning electron microscope (SEM) sample |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1369699A (en) * | 2002-03-06 | 2002-09-18 | 中国科学院水生生物研究所 | Section staining method for tooth of odontoceti |
| CN101390510A (en) * | 2008-11-12 | 2009-03-25 | 中国农业大学 | Preprocessing method of Orthoptera insect integer serial section |
| CN103759994A (en) * | 2014-01-02 | 2014-04-30 | 河南科技大学 | Larva paraffin section forming method |
| CN103954484A (en) * | 2014-04-02 | 2014-07-30 | 浙江万里学院 | Method for producing early gonad paraffin section of Chinese soft shell turtle |
-
2015
- 2015-08-14 CN CN201510505600.8A patent/CN105181406B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1369699A (en) * | 2002-03-06 | 2002-09-18 | 中国科学院水生生物研究所 | Section staining method for tooth of odontoceti |
| CN101390510A (en) * | 2008-11-12 | 2009-03-25 | 中国农业大学 | Preprocessing method of Orthoptera insect integer serial section |
| CN103759994A (en) * | 2014-01-02 | 2014-04-30 | 河南科技大学 | Larva paraffin section forming method |
| CN103954484A (en) * | 2014-04-02 | 2014-07-30 | 浙江万里学院 | Method for producing early gonad paraffin section of Chinese soft shell turtle |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105928752A (en) * | 2016-04-13 | 2016-09-07 | 沈阳大学 | Method for staining paraffin section of adult insect |
| CN105928752B (en) * | 2016-04-13 | 2018-11-20 | 沈阳大学 | A kind of insect imago paraffin section colouring method |
| CN106568632A (en) * | 2016-11-08 | 2017-04-19 | 沈阳农业大学 | Method of converting paraffin slice sample into scanning electron microscope (SEM) sample |
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| Publication number | Publication date |
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| CN105181406B (en) | 2018-04-03 |
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