CN103979210A - Long-term storing method and device of plant sample - Google Patents
Long-term storing method and device of plant sample Download PDFInfo
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- CN103979210A CN103979210A CN201410221407.7A CN201410221407A CN103979210A CN 103979210 A CN103979210 A CN 103979210A CN 201410221407 A CN201410221407 A CN 201410221407A CN 103979210 A CN103979210 A CN 103979210A
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Abstract
The invention discloses a long-term storing method and device of a plant sample. The method comprises the following steps: A, selecting a sample bottle; B, pre-treating the plant sample, namely, removing water content, drying to reach constant weight, and further treating the plant sample by crushing and uniformly mixing; C, feeding sample, namely, feeding the treated sample to the bottom of a sample bottom through a medicine spoon, keeping the sample preservation capacity not greater than 4/5 of the volume of the sample bottle, covering a rubber pad on an opening of the sample bottle, and then tightening a screw cap; D, performing vacuum pumping; E, charging inactive gas like nitrogen or helium; F, sealing, namely, treating a thread cap of the sample bottle and a sample label adhered outside the bottle by wax sealing; G, storing in a shady place, namely, packing the sample bottle into a box, protecting from light with a black bag, adhering the label, and then storing; H, storing the sample again according to steps D to G after the sample is acquired so as to improve the continuous usability of the sample. The device comprises a vacuum pumping device, a sample storing device and an air charging device. The long-term storing method and device of the plant sample have the advantages that the operation is convenient, the difficulty at long-term storage of the plant sample is reduced, and the obtained sample is high in reliability.
Description
Technical field
The technology of the present invention belongs to ecology environment technical field, more specifically relate to a kind of plant sample long-term preservation method, also relate to the long-term device of preserving of a kind of plant sample simultaneously, be suitable for studying long-term preservation method and the device of element circulation, the required plant sample of environmental change.
Background technology
In recent years, climate change becomes the hot issue of world research, and the sample of long-time sequence and analysis are the keys of research climate change.Long term experiment is the important means of the problem in science such as research climate change, variation of ecology and environment, biological evolution, agricultural sustainable development, economics, and the long-term preservation of the plant sample of location test changes most important on the impact of plant for research environment.As the long term experiment at location test station, the Lausanne history of existing 150 ~ 170 years, the advantage of its location test uniqueness is the long-term preservation of test article, and its sample provides splendid material for the circulation of research element, bio-diversity, crop pathogens fashion trend, organic-inorganic pollutant accumulation and trend study.The long-term routine test of China is started late, mainly contain the fertility net that the chemical fertilizer net that starts before and after 1980 and nineteen ninety start, and the Chinese ecosystem research network (CERN) that starts to set up establishment for 1988, china natural resources, the relevant comprehensive important scientific problems research experiment network of ecology environment have been formed, long-term preservation to pedotheque is relatively paid attention to, and preserves comparatively perfect in time series.And for plant sample, especially for element circulation, the required plant sample of environmental change, need equally long-term preservation, but due to the restriction of the store method of plant sample at present, most location tests are less less than preservation or stock on hand, and particularly the plant sample of location test starting stage is especially rare, and the plant sample holding time of location test requires 10,20 years easily, the even longer time, and require the proterties of sample constant, just embody the researching value of its preservation.
For studying the store method of element circulation, the required plant sample of environmental change, mainly contain at present: dry normal temperature preservation method, dry Cryopreservation of Viable ,-80 ℃ of refrigerator Cryopreservation methods and nitrogen ultra low temperature Cryopreservation method etc.Principle is to control that plant sample is in lucifuge, keeps away oxygen, in the environment of low gently dried, make the degraded activity of plant sample in minimum state, thereby reach the object of long-term preservation.
The most normally used is at present dry normal temperature preservation method, dry Cryopreservation of Viable, the method sample moisture-sensitive, go mouldy and the impact of insect pest, in preservation process, also often need the loaded down with trivial details process of frequent airing, dehumidifying etc., these are not only consuming time, and expend a large amount of human and material resources, and take inconvenience, sample contamination risk is larger, that seriously reduces sample for a long time can hold capacity.Ultra-low temperature preservation method, as-80 ℃ of refrigerator Cryopreservation methods and nitrogen ultra low temperature Cryopreservation method, generally needs expensive instrumentation, and equipment cost is high, and the energy consumption in preserving process is also higher, is applicable to the short-term Sample storage of the variable element of a small amount of sample and valence state.And for the Sample storage of sequence in enormous quantities long-time, the method will significantly add the retain costs of large sample, preserve for a long time difficulty large.
Therefore, be necessary to design a kind of plant sample long-term preservation method and device thereof of easy, low consumption.The series of problems such as this equipment that just requires the method to reduce to bring in sample preservation process for a long time, shortage of manpower, do not need special equipment, facilitate sample to take, taking rear sample can continue to preserve, the feature with low, the easy and sustainability of later maintenance cost, thus in element circulation in long-term ecological study, environmental change response research, provide in enormous quantities, long-term between the plant sample of sequence.
Summary of the invention
For the deficiency of above-mentioned existence, the object of the invention is to be to provide a kind of long-term preservation method of plant sample.The plant sample long-term preservation method of design construction convenient operation of the present invention, keeping away the situations such as it makes moist, goes mouldy in preservation process occurs, the plant sample of being simultaneously convenient to test analysis is taken and is again sealed up for safekeeping, has solved the long-term preservation of sample and has again taken the contradiction of sealing up for safekeeping.The method is easy to operate, has simplified the long-term preservation difficulty of plant sample, and the sample reliability of acquisition is high.
Another object of the present invention is the device that has been to provide a kind of long-term preservation of plant sample, this device adopts common glass bottle as the material of main part of Sample storage, bottleneck is with rubber pad and screw top closure with holes, so that premenstrual processing sample deposits in rear to carrying out glass bottle sealing and vacuumizing, being filled with gas is inactive gas, then carries out sealing wax, keeps in Dark Place, this device is made simple and easy, simple to operate, can use for a long time, the chronicity of device and the sample reliability of gained are improved.
To achieve these goals, the present invention adopts following technical measures:
The long-term method of preserving of plant sample, specifically comprises the following steps:
(1) sample bottle is selected: according to the proterties of plant sample, representativeness and Sample storage amount, determine the amount of capacity of vessel, choose not isometric slot glass bottle.
(2) pre-processing of plant sample: complete, dry to constant weight, the larger plant sample of size (length >5 cm) can further be pulverized 40 mesh sieves, mix processing, improve the representativeness of preserving sample, so that sample during test analysis is directly taken, can reduce the loss of sample, increase the utilizability of sample and be easy to again preserve.
(3) setting-out: the sample of handling well is slowly put into sample bottle bottom by spoon, Sample storage amount should not surpass 4/5 of sample bottle volume, and then rubber pad in sample bottle lid, screws screw top closure, and sample bottle outer wall is labelled.
(4) vacuumize: by vacuum pump, sample bottle is vacuumized to 15 ~ 30 seconds, until the interior vacustat of bottle is at 0.04 ~ 0.09 Pa.
(5) be filled with inactive gas (nitrogen or helium): utilize gas filled device to be filled with inactive gas in sample bottle, make after airbag and sample bottle air pressure balance, then the inactive gas that injects 5ml more.Can carry out as required repeatedly vacuumizing for 2 ~ 3 times, gas replenishment process, guarantee the quality of inflation.
(6) sealing: sample bottle thread cap cover and the outer sample that labels of bottle are carried out to sealing wax processing.
(7) shading is preserved: by after sample bottle vanning, then with labelling after the shading of black bag, normal temperature (20 ~ 25 ℃) is preserved or put into reefer (4 ℃) and preserves.
(8) sample carries out the again preservation of 4 ~ 7 steps to sample after taking, and increases the sustainable use of sample.
The long-term device of preserving of plant sample, comprises vacuum extractor, contains sampling device and gas filled device, and vacuum extractor is connected with Sheng sampling device by syringe needle, and gas filled device is connected with Sheng sampling device by syringe needle.It is characterized in that: vacuum extractor comprises vacuum pump and syringe needle, select the vacuum pump of 0.04 ~ 0.09 Pa ultimate pressure according to the requirement of degree of vacuum, the aspirate tube end of vacuum pump is installed syringe needle.Contain sampling device and comprise sample bottle, rubber pad, screw top closure, the material of sample bottle is glass, rubber pad is isometrical in sample bottleneck external diameter, rubber pad is placed in the bottle mouth position of sample bottle, the bottleneck of sample bottle is provided with outside thread, by screw thread fit, makes rubber seal bottleneck with screw top closure when screw top closure screws, screw top closure top is provided with aperture, so that syringe needle inserts rubber pad by aperture, enters sample bottle.Gas filled device comprises airbag, syringe, syringe needle and triple valve (common injector for medical purpose, syringe needle and triple valve), triple valve is flow divider valve, airbag freeing pipe end is connected with the top connection of triple valve valve body, and syringe and syringe needle be the left and right joint of connecting tee valve respectively.Being connected to of vacuum extractor, gas filled device and Sheng sampling device: while vacuumizing, the syringe needle of vacuum extractor pierces through rubber pad and enters sample bottle inside by containing sampling device screw top closure top aperture, complete vacuum after syringe needle pull away sample bottle; The syringe needle of gas filled device pierces through rubber pad and enters sample bottle inside by containing sampling device screw top closure top aperture, complete gas replenishment process after syringe needle pull away sample bottle.
Apparatus of the present invention using method is: sample is put into after sample bottle, at sample bottle bottle mouth position, covered rubber pad, then screw screw top closure, complete bottle process that enters of sample; Vacuum extractor connects sample bottle by syringe needle, syringe needle pierces through rubber pad through screw top closure top aperture, the space of needle tip between rubber pad and sample, then glass bottle is vacuumized, vacustat is extracted rapidly syringe needle during a certain value within the scope of 0.04 ~ 0.09 Pa, completes vacuum; Gas filled device connects the sample bottle that exhausts vacuum by syringe needle, syringe needle pierces through rubber pad through screw top closure top aperture, the space of needle tip between rubber pad and sample, make after airbag and sample bottle air pressure balance, inject again the inactive gass of 5ml more, complete the process that is filled with of inactive gas.
The present invention compared with prior art, has the following advantages and effect:
1) containing sampling device main material is glass, stable in properties and non-contaminated samples makes sample preserve for a long time reliability and improves.
2) sample pre-treatments and vacuumizing, makes plant sample through without moisture, no oxygen treating process, has controlled and can cause it to become qualitative factor.
3) inactive gas being filled with, as protection gas, for plant sample provides a stable environment, suppresses the degradation of plant sample.
4) seal with wax and the further control of shading preserving type can cause the factor of the degraded of plant sample.
5) utilizability is strong, later stage operation, easy maintenance.Sample is taken conveniently, and sample segment can dry to sample bottle, again vacuumize, fill inactive gas process after taking, and increases the operability of the long-term preservation of sample.
6) sample can significantly extend the holding time.Existing store method is due to reasons such as qualitative change, energy consumption height, the holding time of plant sample is generally maintained to 3 ~ 5 years, and the present invention can extend the Sample storage time more than 10 years.
Accompanying drawing explanation
Fig. 1 is the structural representation of the long-term save set of a kind of plant sample.
Wherein: 1-vacuum extractor (parts market is all purchased, common), 2-contains sampling device, and (parts market is all purchased, common), 3-gas filled device (parts are common medical, and market is all purchased), 4-vacuum pump (2XZ series Double-stage rotary vane vacuum pump), 5a, 5b-syringe needle, 6-sample bottle (material is glass), 7-rubber pad, 8-screw top closure, 9-airbag, 10-syringe, 11-triple valve.
The specific embodiment
The plant sample using in following examples is conventional plant sample, and its effect is only to prove the preceence of the inventive method, is not a part for technical solution of the present invention.Obviously, those skilled in the art also can verify technical scheme of the present invention with other plant sample and allied equipment.
Embodiment 1:
This method relates to the store method of the rice sample in a small amount that pulverizes and sieves, mixes, and specifically comprises the following steps:
(1) glass bottle of sample bottle selection of small, as little Dewar bottle (10ml).Be used for holding the good sample of degree of uniformity, reserve capacity is generally 5 ~ 10 g.
(2) pre-processing of rice sample: dry 105 ℃ of half an hour that complete, 75 ℃ to constant weight, pulverized and the usual manner pre-processing sample such as mix after 40 mesh sieves, obtain representational sample to reduce the amount of storage of sample, as rice sample only needs to preserve 5 ~ 10 g, also facilitate directly taking of later stage sample.
(3) setting-out: the rice sample of handling well is slowly put into sample bottle bottom by spoon, and amount of storage is controlled at 4/5 position of bottle, then covers rubber pad at sample bottle mouth position, screws screw top closure, and sample bottle outer wall is labelled, completes setting-out process.
(4) vacuumize: syringe needle pierces through rubber pad through screw top closure top aperture, makes the space of needle tip between sample and rubber pad, open vacuum pump, vacuumize 15 ~ 30 seconds, to negative pressure is constant, be advisable, degree of vacuum is 0.06 Pa, extract rapidly syringe needle, complete vacuum.
(5) be filled with inactive gas (nitrogen or helium): utilize gas filled device to fill inactive gas to sample bottle, syringe needle pierces through rubber pad through screw top closure with holes, make the space of needle tip between sample and rubber pad, after sample bottle and airbag air pressure balance, inject again the inactive gass of 5 ml more.Repeatedly vacuumize, gas replenishment process 2 times, guarantee the quality of inflation.
(6) sealing wax: sample bottle screw top closure and the outer sample labeing of bottle are carried out to sealing wax processing.
(7) preserve: by after sample bottle vanning, then with labelling after the shading of black bag, normal temperature (20 ~ 25 ℃) is preserved or put into reefer (4 ℃) and preserves.
To this, all complete the preservation work of a plant sample, after sample is taken, by 4 ~ 7 steps, sample is sealed up for safekeeping again, increase the sustainable use of sample.
The long-term device of preserving of plant sample, comprises vacuum extractor 1, contains sampling device 2 and gas filled device 3.Vacuum extractor 1 comprises vacuum pump 4 and syringe needle 5a, and the aspirate tube end of vacuum pump 4 is installed syringe needle 5a; Contain sampling device 2 and comprise sample bottle 6, rubber pad 7, screw top closure 8, the material of sample bottle 6 is glass, rubber pad 7 is isometrical in the bottleneck external diameter of sample bottle 6, rubber pad 7 is placed in the bottle mouth position of sample bottle 6, the bottleneck of sample bottle 6 is provided with outside thread and screw top closure 8 passes through screw thread fit, screw top closure 8 makes rubber pad 7 sealing bottlenecks while screwing, screw top closure 8 tops are provided with aperture, so that syringe needle 5a pierces through rubber pad 7 through screw top closure 8 top apertures, enter sample bottle 6; Gas filled device 3 comprises airbag 9, syringe 10, triple valve 11 and syringe needle 5b, and triple valve 11 is flow divider valve, and airbag 9 freeing pipe ends are connected with the top connection of triple valve 11 valve bodies, and syringe 10 and syringe needle 5b be the left and right joint of connecting tee valve 11 respectively.Vacuum extractor 1, gas filled device 3 are connected to Sheng sampling device 2: while vacuumizing, the syringe needle 5a of vacuum extractor 1 pierces through rubber pad 7 by the aperture on screw top closure 8 tops in Sheng sampling device 2 and enters sample bottle 6 inside, make the space of syringe needle 5a tip between rubber pad 7 and sample, complete vacuum after syringe needle 5a pull away sample bottle 6; Gas filled device 2 connects the sample bottle 6 that exhausts vacuum by syringe needle 5b, syringe needle 5b pierces through rubber pad 7 through screw top closure 8 top apertures, make the space of syringe needle 5b tip between rubber pad 7 and sample, complete the process that is filled with of inactive gas after syringe needle 5b pull away sample bottle 6.
Embodiment 2:
This method, for the store method of the relatively large straw sample that pulverizes and sieves, mixes, specifically comprises the following steps:
(1) sample bottle is selected the slot glass bottle of 250 ml, because straw plant individuality is larger, need to select the above straw of 3 root and stem of certain plants to pulverize to obtain representative sample, preserves sample size generally more than 50g.
(2) pre-processing of straw sample: dry 105 ℃ of half an hour that complete, 75 ℃ to constant weight, pulverized and the usual manner pre-processing sample such as mix after 40 mesh sieves, obtain representational sample.
(3) setting-out: the straw sample of handling well is slowly put into sample bottle bottom by spoon, and amount of storage is controlled at 4/5 position of bottle, then covers rubber pad at sample bottle mouth position, screws screw top closure, and sample bottle outer wall is labelled, completes setting-out process.
(4) vacuumize: syringe needle pierces through rubber pad through screw top closure top aperture, makes the space of needle tip between sample and rubber pad, open vacuum pump, vacuumize 15 ~ 30 seconds, to negative pressure is constant, be advisable, degree of vacuum is 0.05 Pa, extract rapidly syringe needle, complete vacuum.
(5) be filled with inactive gas (nitrogen or helium): utilize gas filled device to fill inactive gas to sample bottle, syringe needle pierces through rubber pad through screw top closure top aperture, make the space of needle tip between sample and rubber pad, after sample bottle and airbag air pressure balance, inject again the inactive gass of 5 ml more.Repeatedly vacuumize, gas replenishment process 3 times, guarantee the quality of inflation.
(6) sealing wax: sample bottle screw top closure and the outer sample labeing of bottle are carried out to sealing wax processing.
(7) preserve: by after sample bottle vanning, then with labelling after the shading of black bag, normal temperature (20 ~ 25 ℃) is preserved or put into reefer (4 ℃) and preserves.
To this, all complete the preservation work of a plant sample, after sample is taken, by 4 ~ 7 steps, sample is sealed up for safekeeping again, increase the sustainable use of sample.
Claims (2)
1. the long-term method of preserving of plant sample, the steps include:
A, sample bottle are selected: according to the proterties of plant sample, representativeness and Sample storage amount, determine the amount of capacity of vessel, choose not isometric slot glass bottle;
The pre-processing of B, plant sample: complete, dry to constant weight, processing is further pulverized, mixed to plant sample;
C, setting-out: the sample of handling well is put into sample bottle bottom by spoon, and Sample storage amount is no more than 4/5 of sample bottle volume, and rubber pad in sample bottle lid, screws screw top closure, and sample bottle outer wall is labelled;
D, vacuumize: by vacuum pump, sample bottle is vacuumized to 15 ~ 30 seconds, until in bottle vacustat at 0.04 ~ 0.09 Pa;
E, be filled with inactive gas: nitrogen or helium, utilize gas filled device to be filled with inactive gas in sample bottle, make after airbag and sample bottle air pressure balance, the inactive gas of the 5ml that reinjects, vacuumizes for 2-3 time, gas replenishment process repeatedly;
F, sealing: sample bottle thread cap cover and the outer sample labeing of bottle are carried out to sealing wax processing;
G, shading are preserved: by after sample bottle vanning, then with labelling after the shading of black bag, 4 ℃ of preservations of reefer are preserved or put into normal temperature;
H, sample re-start the preservation again of sample by D ~ G step after taking.
2. the device that a kind of plant sample claimed in claim 1 is preserved for a long time, comprise vacuum extractor (1), contain sampling device (2) and gas filled device (3), vacuum extractor (1) is connected with Sheng sampling device (2) by syringe needle (5a), gas filled device (3) is connected with Sheng sampling device (2) by syringe needle (5b), it is characterized in that: vacuum extractor (1) comprises vacuum pump (4) and syringe needle (5a), the aspirate tube end of vacuum pump (4) is installed syringe needle (5a); Contain sampling device (2) and comprise sample bottle (6), rubber pad (7), screw top closure (8), the material of sample bottle (6) is glass, rubber pad (7) is isometrical in the bottleneck external diameter of sample bottle (6), rubber pad (7) is placed in the bottle mouth position of sample bottle (6), the bottleneck of sample bottle (6) is provided with outside thread and screw top closure (8) passes through screw thread fit, screw top closure (8) screws and makes rubber pad (7) sealing bottleneck, and screw top closure (8) top is provided with aperture; Gas filled device (3) comprises airbag (9), syringe (10), triple valve (11) and syringe needle (5b), triple valve (11) is flow divider valve, airbag (9) freeing pipe end is connected with the top connection of triple valve (11) valve body, syringe (10) and syringe needle (5b) be the left and right joint of connecting tee valve (11) respectively, and syringe needle (5a) or syringe needle (5b) pierce through rubber pad (7) through screw top closure (8) top aperture and enter sample bottle (6).
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| CN201410221407.7A CN103979210B (en) | 2014-05-23 | 2014-05-23 | The long-term preservation method of a kind of plant sample and device |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106672441A (en) * | 2017-01-23 | 2017-05-17 | 浙江丽生生物科技有限公司 | DNA specimen normal temperature preservation preparation method and packaging equipment |
| CN107588995A (en) * | 2016-07-06 | 2018-01-16 | 中国石油化工股份有限公司 | A kind of adsorption equilibrium tank and its application |
| CN111846625A (en) * | 2020-07-04 | 2020-10-30 | 江苏汭博医疗科技有限公司 | Biological sample storage device |
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