CN105476646A - Method for efficiently extracting hemolymph of bivalve - Google Patents
Method for efficiently extracting hemolymph of bivalve Download PDFInfo
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- CN105476646A CN105476646A CN201610006424.8A CN201610006424A CN105476646A CN 105476646 A CN105476646 A CN 105476646A CN 201610006424 A CN201610006424 A CN 201610006424A CN 105476646 A CN105476646 A CN 105476646A
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- bivalve
- hemolymph
- bivalve shellfish
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/15—Devices for taking samples of blood
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D3/00—Appliances for supporting or fettering animals for operative purposes
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Abstract
The invention relates to a method for efficiently extracting hemolymph of bivalve. The method includes the following steps that a, seawater on the surface of the bivalve is completely absorbed through cotton balls, the surface of the bivalve is wiped with ethyl alcohol to be sterilized, and seawater on the surface of the bivalve is completely sucked up through cotton balls; b, the bivalve which is completely wiped is placed in a groove of an extracting device, the adductor muscle of the bivalve is ripped open, a shell is open, and two shells are flatly open and erected in the groove; c, after pelecypod of the bivalve and the portion below a water pipe are cushioned with cotton balls, an outer sleeve membrane above a pericardial cavity is slightly shifted away through tweezers, and the pericardial cavity is completely exposed; d, a gun head of a liquid moving device is cut, meanwhile, the pelecypod of the bivalve is stimulated, and hemolymph is fast sucked with the liquid moving device when the heart beats and the pericardial cavity swells. The method has the advantages that the method is easy to operate, the total amount of the extracted hemolymph of the bivalve can be effectively increased, the purity of the hemolymph is improved, and the operating environment can be kept clean in the operating process.
Description
[technical field]
The present invention relates to the extractive technique field of bivalve shellfish hemolymph, specifically, is a kind of extracting method of efficient bivalve shellfish hemolymph.
[background technology]
Nowadays be the epoch of ocean, culture fishery develops rapidly.Bivalve shellfish is as important economy aquatic animals, and in production practices, disease is the key factor of high yield always, and studied person paid close attention to always.And hemolymph is as the important immuning tissue of shellfish, there is high researching value.
Due to bivalve generic open pipe formula blood circulation, the collection of its hemolymph is difficult point always.At present, researcher multi injector and syringe needle, but traditional to get the method syringe needle of blood with syringe and syringe needle meticulous, easily blocks during absorption, often cause the amount drawn inadequate, and be easily extracted into body fluid and impure.
Based on above-mentioned reason, in the leaching process of hemolymph, purity is not high, amount is few is the problem perplexing researcher always.Therefore, people wish the method finding a large amount of and pure shellfish hemolymph of a kind of simple and effective extraction.
[summary of the invention]
The object of the invention is for deficiency of the prior art and above-mentioned problem demanding prompt solution, the extracting method of bivalve shellfish hemolymph of a kind of simple operation, hygiene high-effective is provided
For achieving the above object, the technical scheme that the present invention takes is: a kind of extracting method of efficient bivalve shellfish hemolymph, comprise the following steps: the sea water on a. cotton balls exhaustion bivalve shellfish surface, and carry out disinfection with its surface of alcohol swab wiping, again the sea water on bivalve shellfish surface is drawn clean with cotton balls; B. the bivalve shellfish of wiped clean is vertically stood in the groove of extraction element, the anterior closed shell flesh of bivalve shellfish is scratched, makes dried liquid stream in bivalve shellfish body; C. bivalve shellfish to be inverted in groove and to be strutted, cutting off its rear closed shell flesh, shell is opened, making that bivalve is open and flat to be stood within groove; D. puncturing body cavity makes coelomic fluid residual in body flow out, and with cotton balls, the body fluid of Sinonovacula constricta tissue surface is blotted, after then padding cotton balls below bivalve shellfish abdominal foot and water pipe, with tweezers the outer embrane above pricardial coelom dial gently from, pricardial coelom is exposed completely; E. cut the rifle head of pipettor, stimulate the abdominal foot of bivalve shellfish simultaneously, draw rapidly with pipettor when heartbeat pricardial coelom is heaved.
Again with cotton balls, the sea water on bivalve shellfish surface being drawn clean object in described step a is the degree of purity ensureing to draw hemolymph.
Below bivalve shellfish abdominal foot and water pipe, pad cotton balls for draw body fluid in described steps d, fix bivalve shellfish abdominal foot and water pipe simultaneously, prevent abdominal foot irriate and slide.
When cutting the rifle head of pipettor in described step e, cut away the tip of rifle head portion 3-4mm to increase the bore diameter of rifle head.
During described step e moderate stimulation abdominal foot, bivalve shellfish hemolymph can be back in pricardial coelom, and assembles in a large number in pricardial coelom, and pricardial coelom is heaved, and pricardial coelom film is thinning.
The invention has the advantages that:
1, the inventive method is simple to operate, and effectively can increase the total amount of the bivalve shellfish hemolymph extracted, and promotes the purity of hemolymph, can keep the clean and tidy of operating environment in operating process.
2, repeatedly use cotton balls auxiliary operation in the inventive method, support on the one hand the attitude that bivalve shellfish makes it be in be beneficial to extraction, the surplus fluid that can produce in absorption operation process in time on the one hand, and cotton balls easily gets convenience, with low cost.
3, the inventive method is when using pipettor, is cut by the tip of pipettor gun head, avoids rifle head to block the total amount having influence on hemolymph extraction.
4, the inventive method stimulated the abdominal foot of bivalve shellfish before extracting hemolymph, and hemolymph is gathered in pricardial coelom in a large number, provided purity and the total amount of hemolymph on the one hand, made pricardial coelom expansion heave pricardial coelom film on the one hand thinning, was easy to broken by pipettor suction.
[accompanying drawing explanation]
Accompanying drawing 1 is the inventive method operating means schematic diagram.
Accompanying drawing 2 is plastic board schematic diagrams of the inventive method operating means.
Accompanying drawing 3 is pipettor gun head cut place schematic diagrams.
[detailed description of the invention]
Below in conjunction with accompanying drawing, detailed description of the invention provided by the invention is elaborated.In explanation, the bivalve shellfish in the inventive method carries out the extraction of hemolymph to it for Sinonovacula constricta.
The Reference numeral related in accompanying drawing and ingredient as follows:
1. metal framework 2. plastic board 3. drawer-like capsule 4. handle
5. groove 6. aperture 7. rifle head cut place.
First to exhaust supporting the Sinonovacula constricta cotton balls after 2-3 days temporarily sea surface, then carry out disinfection with alcohol swab wipe surfaces, again with cotton balls, the sea water of Sinonovacula constricta tissue surface is drawn totally, be placed in the groove 5 of extraction element (as shown in Figure 1).Again with cotton balls, the sea water of Sinonovacula constricta tissue surface being drawn clean object is herein the hemolymph purity ensureing further to draw.Here also it should be noted that, should by the front of Sinonovacula constricta (the morphologic front of shellfish) down during placement Sinonovacula constricta, back of the body side (the morphologic back of shellfish) is vertically placed towards observer and is stood in groove 5.Then from the back of the body side of Sinonovacula constricta, hinge forwards to be drawn with No. 23 dissecting knife and disconnected the closed shell flesh of Sinonovacula constricta front portion to be scratched.
Have a large amount of body fluid after scratching the closed shell flesh of Sinonovacula constricta front portion to flow out, after body fluid drains off, Sinonovacula constricta to be inverted in this device groove 5 and slightly to strut (can not overexert, can know the boundary between resolution outer embrane and shell) with hands or pliers (large individual bivalve shellfish uses).Then with dissecting knife prolong edge between outer embrane and shell gently down (i.e. the morphologic rear of shellfish) scratch (must guard against near pricardial coelom and scratch pricardial coelom), until cut off rear closed shell flesh.Then slowly remove one side shell of bivalve shellfish, shell is opened, make that bivalve is open and flat to be stood within device groove 5.Drawing has no progeny goes out having a large amount of body of shellfish liquid stream, and now continuation dissecting knife scratches body cavity coelomic fluid residual in body is fully flowed out.Here it should be noted that, have a large amount of body fluid to flow out when carrying out aforesaid operations, these body fluid can be flowed to by the cylindric aperture 6 in extraction element groove in the plastic drawer shape capsule 3 of bottom of device (as shown in Figure 2), plastic drawer shape capsule 3 is passable device, when its inner body fluid is too much, extract plastic drawer shape capsule 3 out by its front handle 4 body fluid is abandoned, reusable after cleaning.
After fully flowing out at Sinonovacula constricta body fluid, with cotton balls, the body fluid of Sinonovacula constricta tissue surface is blotted, and pad appropriate cotton balls below Sinonovacula constricta abdominal foot and water pipe.The object padding appropriate cotton balls below abdominal foot and water pipe draws totally by the body fluid of shellfish further, and ensure hemolymph purity, and play certain fixation to abdominal foot and water pipe, so that when stimulating abdominal foot, abdominal foot does not slide.Then being taken off by the moveable plastic board 2 of device lies against in laboratory table, to needle outer embrane above pricardial coelom and pricardial coelom film along the auricular edge of Sinonovacula constricta with tweezers (9cm fine dissection tip tweezers) and it is dialled gently from, pricardial coelom is fully come out, be convenient to the absorption of pipettor suction to hemolymph, ensure that heart is not punctured.
Finally the rifle head of 1000ul pipettor is cut away about 3-4mm (as shown in Figure 3, being approximately the size of heart) from its rifle head cut place 7, stimulate Sinonovacula constricta abdominal foot, hemolymph is back in pricardial coelom, and assembles in a large number in pricardial coelom.When heartbeat pricardial coelom is heaved, draw hemolymph rapidly with pipettor.Here it should be noted that, adopting the rifle head of 1000ul pipettor in order to ensure that pricardial coelom is enough inhaled broken by suction, hemolymph being sucked in rifle head.In accompanying drawing 3, rifle head being cut away 3-4mm (being approximately the size of heart) tip, is the bore diameter in order to increase rifle head, prevents the heart blocking bore shrinking rear bulk.Stimulate the object of abdominal foot to be make hemolymph assemble the extracted amount improving hemolymph on the one hand, pricardial coelom is heaved on the other hand, and pricardial coelom film is thinning, is inhaled by pricardial coelom broken when being conducive to the absorption with pipettor.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the inventive method; can also make some improvement and supplement, these improve and supplement and also should be considered as protection scope of the present invention.
Claims (5)
1. the extracting method of an efficient bivalve shellfish hemolymph, it is characterized in that, described method comprises the following steps: the sea water on a. cotton balls exhaustion bivalve shellfish surface, and carries out disinfection with its surface of alcohol swab wiping, again the sea water on bivalve shellfish surface is drawn clean with cotton balls; B. the bivalve shellfish of wiped clean is vertically stood in the groove of extraction element, the anterior closed shell flesh of bivalve shellfish is scratched, makes dried liquid stream in bivalve shellfish body; C. bivalve shellfish to be inverted in groove and to be strutted, cutting off its rear closed shell flesh, shell is opened, making that bivalve is open and flat to be stood within groove; D. puncturing body cavity makes coelomic fluid residual in body flow out, and with cotton balls, the body fluid of Sinonovacula constricta tissue surface is blotted, after then padding cotton balls below bivalve shellfish abdominal foot and water pipe, with tweezers the outer embrane above pricardial coelom dial gently from, pricardial coelom is exposed completely; E. cut the rifle head of pipettor, stimulate the abdominal foot of bivalve shellfish simultaneously, draw rapidly with pipettor when heartbeat pricardial coelom is heaved.
2. the extracting method of a kind of efficient bivalve shellfish hemolymph according to claim 1, is characterized in that, again with cotton balls, the sea water on bivalve shellfish surface being drawn clean object in described step a is the degree of purity ensureing to draw hemolymph.
3. the extracting method of a kind of efficient bivalve shellfish hemolymph according to claim 1, it is characterized in that, below bivalve shellfish abdominal foot and water pipe, pad cotton balls for draw body fluid in described steps d, fix bivalve shellfish abdominal foot and water pipe simultaneously, prevent abdominal foot irriate and slide.
4. the extracting method of a kind of efficient bivalve shellfish hemolymph according to claim 1, is characterized in that, when cutting the rifle head of pipettor in described step e, cuts away the tip of rifle head portion 3-4mm to increase the bore diameter of rifle head.
5. the extracting method of a kind of efficient bivalve shellfish hemolymph according to claim 1, is characterized in that, during described step e moderate stimulation abdominal foot, bivalve shellfish hemolymph can be back in pricardial coelom, and assemble in a large number in pricardial coelom, pricardial coelom is heaved, and pricardial coelom film is thinning.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610006424.8A CN105476646B (en) | 2016-01-06 | 2016-01-06 | A kind of extracting method of efficient bivalve shellfish hemolymph |
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| CN201610006424.8A CN105476646B (en) | 2016-01-06 | 2016-01-06 | A kind of extracting method of efficient bivalve shellfish hemolymph |
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| CN105476646A true CN105476646A (en) | 2016-04-13 |
| CN105476646B CN105476646B (en) | 2019-04-09 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107817259A (en) * | 2017-10-30 | 2018-03-20 | 中国水产科学研究院黄海水产研究所 | A kind of method of seawater blood clam section shellfish blood cell collection and Ultrastructural observation |
| CN110794024A (en) * | 2019-11-30 | 2020-02-14 | 中国海洋大学 | Standardized determination method for shellfish blood physiological indexes |
| CN112608825A (en) * | 2020-12-28 | 2021-04-06 | 浙江省海洋水产研究所 | Visual mussel hemolymph collection system |
| CN115125185A (en) * | 2022-08-30 | 2022-09-30 | 中国海洋大学 | Preparation method of bivalve blood lymphocyte single cell suspension |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1598583A (en) * | 2004-08-20 | 2005-03-23 | 国家海洋局第一海洋研究所 | Testing method of stream cell operation of heat shock protein 70 in scallop blood lymphocyte |
| CN102321578A (en) * | 2011-07-13 | 2012-01-18 | 中国水产科学研究院东海水产研究所 | Purification method of marine bivalve shellfish blood cell |
-
2016
- 2016-01-06 CN CN201610006424.8A patent/CN105476646B/en active Active
Patent Citations (2)
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| CN1598583A (en) * | 2004-08-20 | 2005-03-23 | 国家海洋局第一海洋研究所 | Testing method of stream cell operation of heat shock protein 70 in scallop blood lymphocyte |
| CN102321578A (en) * | 2011-07-13 | 2012-01-18 | 中国水产科学研究院东海水产研究所 | Purification method of marine bivalve shellfish blood cell |
Non-Patent Citations (1)
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| 高菲 等: "双壳贝类血淋巴中儿茶酚胺的检测方法初步研究", 《海洋科学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107817259A (en) * | 2017-10-30 | 2018-03-20 | 中国水产科学研究院黄海水产研究所 | A kind of method of seawater blood clam section shellfish blood cell collection and Ultrastructural observation |
| CN110794024A (en) * | 2019-11-30 | 2020-02-14 | 中国海洋大学 | Standardized determination method for shellfish blood physiological indexes |
| CN110794024B (en) * | 2019-11-30 | 2022-05-10 | 中国海洋大学 | Standardized determination method for shellfish blood physiological indexes |
| CN112608825A (en) * | 2020-12-28 | 2021-04-06 | 浙江省海洋水产研究所 | Visual mussel hemolymph collection system |
| CN112608825B (en) * | 2020-12-28 | 2023-12-12 | 浙江省海洋水产研究所 | Visual mussel blood lymph collection system |
| CN115125185A (en) * | 2022-08-30 | 2022-09-30 | 中国海洋大学 | Preparation method of bivalve blood lymphocyte single cell suspension |
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| CN105476646B (en) | 2019-04-09 |
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