CN102268366A - Metabonomic cell rapid sampling quenching instrument - Google Patents
Metabonomic cell rapid sampling quenching instrument Download PDFInfo
- Publication number
- CN102268366A CN102268366A CN2011101720449A CN201110172044A CN102268366A CN 102268366 A CN102268366 A CN 102268366A CN 2011101720449 A CN2011101720449 A CN 2011101720449A CN 201110172044 A CN201110172044 A CN 201110172044A CN 102268366 A CN102268366 A CN 102268366A
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- Prior art keywords
- cell
- rotating cylinder
- taking
- district
- metabolism group
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- 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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
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- 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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
- C12M33/06—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles for multiple inoculation or multiple collection of samples
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- 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
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/02—Separating microorganisms from the culture medium; Concentration of biomass
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- 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
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/14—Drying
Abstract
The invention discloses a metabonomic cell rapid sampling quenching instrument, which is characterized in that: the instrument comprises a filtering rotating cylinder for collecting cell culture; the filtering rotating cylinder comprises a filtering blotting-up zone, a cell washing zone, and a discharging zone; the filtering blotting-up zone and the cell washing zone are connected to a vacuum pipe respectively; the discharging zone is connected to a compressed air pipe; a cell washing solution nozzle is disposed outside the cell washing zone; a freezing scraper which can scrape the cell culture on the filtering rotating cylinder is disposed outside the discharging zone; one end of the freezing scraper is immersed in a freezing source. The purpose of the invention is to overcome the disadvantages in the prior art, and to provide a metabonomic cell rapid sampling quenching instrument which has a simple structure, can automatically complete whole-process operations of sampling, washing, and quenching, and is convenient for controlling the period of the quenching process within a scope as short as possible.
Description
Technical field
The present invention relates to a kind of metabolism group cell cancellation instrument of taking a sample fast.
Background technology
Collecting cell from liquid such as microbial cells sampling, cell sampling, when carrying out correlative studys such as metabolism group or Physiology and biochemistry, need be at moment washed cell and the Metabolic activity of cancellation cell, in level or to finish this operation in the submicrosecond level time most important second, be related to the variation of endocellular metabolism thing group or some molecular structure, concentration or kind, this fast process of sampling be intended to moment " freezing " physiological status of cells, reduce as much as possible from sampling and begin when Metabolic activity stops fully during this period of time.
Common way has at present:
1, from cell or microbial culture medium, gets liquid and carry out the centrifugal collecting precipitation cell, add cryogenic fluid buffered soln, physiological saline or methanol solution etc. about 4 ℃ then and carry out several times the operation of " cell washing-centrifugation ", to remove remaining interfering substances such as substratum, clean cell and utilize cryogenic fluid, dry ice or liquid nitrogen to carry out the Metabolic activity of quick-frozen cancellation cell more afterwards.The required formality of this process is very numerous and diverse, and maximum drawback is that the whole process time of cancellation that is sampled to is often between 10～30 minutes, can't " freeze " Metabolic activity of some biochemical reaction speed at all, also cannot guarantee the physiological status the when sample that is finally obtained can keep sampling at all in the submicrosecond level.The drawback of this sampling method becomes the bottleneck technology of microorganism or the research of cellular metabolism group.
2 utilize the syringe that refrigerant low-temperature methanol solution or freezing steel ball are housed directly to draw the nutrient solution sampling, carry out the low-temperature centrifugation collecting cell then, the benefit of doing like this is, can the sampling in moment freeze physiological status of cells, but greatest drawback is, because cell is earlier by cancellation and then wash, in fact the cancellation process can cause the cell walls damaged and break in a large number, in subsequently washing-centrifugal or filter in the collection process, a large amount of leaking can be taken place and be caused accurately obtaining the concentration and the kind of information of various molecules in the cell in intracellular material.Therefore, this method also is that limitation is arranged very much in microorganism or the research of cellular metabolism group.
3 utilize millipore filtration to filter nutrient solution fast, are immersed into then in-40 ℃ the frozen soln to carry out the cancellation of Metabolic activity, and then carry out the filtration and the washing of cell, and that is set forth in the limitation of this method and 2 is consistent.
4 utilize millipore filtration to filter nutrient solution fast, utilize normal temperature solution to carry out filtering while washing fast then, operate three times, and the cell that scrapes on the filter membrane carries out the liquid nitrogen cancellation, and this working method can be with whole process control within 15 seconds.Though exist the limitation that does not have timely cancellation Metabolic activity, be present reliable feasible method fast.
Summary of the invention
The objective of the invention is in order to overcome weak point of the prior art, provide a kind of simple in structure, can automatization ground finish sampling, washing, the operation of cancellation whole process, be convenient to sampling cancellation process time is controlled at metabolism group cell in the scope of lacking the as far as possible cancellation instrument of taking a sample fast.
In order to achieve the above object, the present invention adopts following scheme:
A kind of metabolism group cell cancellation instrument of taking a sample fast, it is characterized in that: but comprise the filtration rotating cylinder of collecting cell culture, described filtration rotating cylinder comprises filtering and blots the district, cell washing district and discharge zone, described filtration is blotted the district and is connected with valve tube respectively with the cell washing district, described discharge zone and pressurized air pipe coupling, be provided with cell washing solution shower nozzle in the outside, described cell washing district, be provided with outside described discharge zone and can will filter the freezing scraper that cell culture scrapes on the rotating cylinder, described freezing scraper one end is immersed in the refrigerated source.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast is characterized in that described filtration rotating cylinder comprises the stationary cylinder and the rotating cylinder of concentric setting.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast, it is characterized in that in described stationary cylinder, blotting the district with described filtration, be respectively equipped with the imbibition groove on the position of cell washing district and discharge zone correspondence, suction groove and vent grooves, described imbibition groove, the suction groove is connected with vacuum extractor respectively, and described vent grooves is connected with pneumatic plant.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast is characterized in that described filtration rotating cylinder is divided into 18 fan sections, and wherein 1 to 9 district blots the district for filtering, and 10 to 16 districts are the cell washing district, and 17 and 18 districts are discharge zone.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast is characterized in that being provided with sample and collects plate on described freezing scraper.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast is characterized in that being provided with filter element on described filtration rotating cylinder.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast is characterized in that described filter element is the millipore filter that materials such as cylindric millipore filtration or stainless steel are made.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast is characterized in that also comprising that the microorganism of dispersing or cell can be inhaled the air of considering inhales and consider device.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast is characterized in that described refrigerated source is a liquid nitrogen container, a kind of in refrigerant condenser, dry ice or the cryogenic freezing liquid.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast is characterized in that described filtration rotating cylinder is a kind of in single cylinder, twin-roll or the multiple-roll.
Aforesaid a kind of metabolism group cell cancellation instrument of taking a sample fast, the cross section that it is characterized in that described filtration rotating cylinder be for circular, a kind of in oblong or the flat drum shape.
In sum, the present invention with respect to its beneficial effect of prior art is:
The present invention can finish to automatization sampling, washing, the operation of cancellation whole process, be convenient to sampling cancellation process time is controlled in the short as far as possible scope, time that the soonest can only 15 seconds in the time of can be with manual operation reduces 10 seconds again, just can finish entire operation in 5 seconds, thereby reduce the variation of molecular species, concentration and structure in the cell in sampling cancellation process, almost reaching moment " freezes " effect of the biochemical state of cell physiological.
Description of drawings
Fig. 1 is a synoptic diagram of the present invention;
Fig. 2 is the synoptic diagram of stationary cylinder.
Embodiment
Below in conjunction with description of drawings and embodiment the present invention is further described:
The cancellation instrument of taking a sample fast of a kind of metabolism group cell as illustrated in fig. 1 and 2, but the filtration rotating cylinder 10 that comprises the collecting cell culture, filtration rotating cylinder 10 described in the process of sampling is arranged in the sampling receptacle 90 that is mounted with microorganism goods cell culture, described filtration rotating cylinder 10 is connected with drive unit, under the effect of drive unit, filtering rotating cylinder 10 can rotate, and adjustable-speed, and wherein said drive unit can be a motor; Described filtration rotating cylinder 10 comprises filtering to blot distinguishes 20, cell washing district 30 and discharge zone 40, described filtration is blotted district 20 and is connected with valve tube respectively with cell washing district 30, described discharge zone 40 and pressurized air pipe coupling, be provided with cell washing solution shower nozzle 50 in 30 outsides, described cell washing district, be provided with outside described discharge zone 40 and can will filter the freezing scraper 60 that cell culture scrapes on the rotating cylinder 10, described freezing scraper 60 1 ends are immersed in liquid nitrogen and irritate in 70.Freezing scraper 60 has thermal conductive resin, and the one end is immersed in liquid nitrogen and irritates in the liquid nitrogen in 70, guarantees temperature on the freezing scraper 60 for below-60 ℃, freezes the effect of cell, its Metabolic activity of cancellation to reach moment.Wherein said freezing scraper 60 also can be immersed in refrigerant condenser, dry ice or the cryogenic freezing liquid.
Filtration rotating cylinder (10) described in the present invention can be a kind of in single cylinder, twin-roll or the multiple-roll.The cross section of described filtration rotating cylinder (10) is circular, a kind of in oblong or the flat drum shape.
On described freezing scraper 60, be provided with sample among the present invention and collect plate 80.The cell that scrapes off obtains collecting and keeping always the sharp freezing state at collection plate place.
On described filtration rotating cylinder 10, be provided with filter element among the present invention.Described filter element is the millipore filter that materials such as cylindric millipore filtration or stainless steel are made.
Also comprise among the present invention and the microorganism or the cell that disperse can be inhaled the air suction worry device of considering.Effectively guarantee Biosafety.
The present invention can finish to automatization sampling, washing, the operation of cancellation whole process, be convenient to sampling cancellation process time is controlled in the short as far as possible scope, time that the soonest can only 15 seconds in the time of can be with manual operation reduces 10 seconds again, just can finish entire operation in 5 seconds, thereby reduce the variation of molecular species, concentration and structure in the cell in sampling cancellation process, almost reaching moment " freezes " effect of the biochemical state of cell physiological.
Claims (10)
1. metabolism group cell cancellation instrument of taking a sample fast, it is characterized in that: but comprise the filtration rotating cylinder (10) of collecting cell culture, described filtration rotating cylinder (10) comprises filtering and blots district (20), cell washing district (30) and discharge zone (40), described filtration is blotted district (20) and is connected with valve tube respectively with cell washing district (30), described discharge zone (40) and pressurized air pipe coupling, be provided with cell washing solution shower nozzle (50) in the outside, described cell washing district (30), be provided with outside described discharge zone (40) and can go up the freezing scraper (60) that cell culture scrapes with filtering rotating cylinder (10), described freezing scraper (60) one ends are immersed in the refrigerated source (70).
2. a kind of metabolism group cell according to claim 1 cancellation instrument of taking a sample fast is characterized in that described filtration rotating cylinder (10) comprises the stationary cylinder (101) and the rotating cylinder (102) of concentric setting.
3. a kind of metabolism group cell according to claim 2 cancellation instrument of taking a sample fast, it is characterized in that in described stationary cylinder (101), blotting district (20) with described filtration, be respectively equipped with imbibition groove (201) on the position of cell washing district (30) and discharge zone (40) correspondence, suction groove (301) and vent grooves (401), described imbibition groove (201), suction groove (301) is connected with vacuum extractor respectively, and described vent grooves (401) is connected with pneumatic plant.
4. according to claim 1 or the 2 or 3 described a kind of metabolism group cells cancellation instrument of taking a sample fast, it is characterized in that described filtration rotating cylinder (10) is divided into 18 fan sections (1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18), wherein 1 to 9 district blots district (20) for filtering, and 10 to 16 districts are cell washing district (30), and 17 and 18 districts are discharge zone (40).
5. a kind of metabolism group cell according to claim 1 cancellation instrument of taking a sample fast is characterized in that being provided with sample and collects plate (80) on described freezing scraper (60).
6. a kind of metabolism group cell according to claim 1 cancellation instrument of taking a sample fast is characterized in that being provided with filter element on described filtration rotating cylinder (10).
7. a kind of metabolism group cell according to claim 1 cancellation instrument of taking a sample fast is characterized in that also comprising that the microorganism of dispersing or cell can be inhaled the air of considering inhales and consider device.
8. a kind of metabolism group cell according to claim 1 cancellation instrument of taking a sample fast is characterized in that described refrigerated source is a liquid nitrogen container, a kind of in refrigerant condenser, dry ice or the cryogenic freezing liquid.
9. a kind of metabolism group cell according to claim 1 cancellation instrument of taking a sample fast is characterized in that a kind of in single cylinder, twin-roll or the multiple-roll of described filtration rotating cylinder (10).
10. according to claim 1 or the 2 or 3 described a kind of metabolism group cells cancellation instrument of taking a sample fast, the cross section that it is characterized in that described filtration rotating cylinder (10) is for circular, a kind of in oblong or the flat drum shape.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110172044 CN102268366B (en) | 2011-06-24 | 2011-06-24 | Metabonomic cell rapid sampling quenching instrument |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110172044 CN102268366B (en) | 2011-06-24 | 2011-06-24 | Metabonomic cell rapid sampling quenching instrument |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102268366A true CN102268366A (en) | 2011-12-07 |
| CN102268366B CN102268366B (en) | 2013-04-17 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110172044 Expired - Fee Related CN102268366B (en) | 2011-06-24 | 2011-06-24 | Metabonomic cell rapid sampling quenching instrument |
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| Country | Link |
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| CN (1) | CN102268366B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754337A (en) * | 2016-12-09 | 2017-05-31 | 大连工业大学 | A kind of cell metabolism quenching system and its cell metabolism quenching method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07147969A (en) * | 1993-11-27 | 1995-06-13 | Yamasa Shoyu Co Ltd | Tool for transporting cell and method for transporting cell using the same |
| CN1157634A (en) * | 1994-09-13 | 1997-08-20 | 卡尔·托马博士公司 | Novel sampling system for use in analysis of biological processes |
| CN202246689U (en) * | 2011-06-24 | 2012-05-30 | 吴小禾 | Sampling and quenching equipment |
-
2011
- 2011-06-24 CN CN 201110172044 patent/CN102268366B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07147969A (en) * | 1993-11-27 | 1995-06-13 | Yamasa Shoyu Co Ltd | Tool for transporting cell and method for transporting cell using the same |
| CN1157634A (en) * | 1994-09-13 | 1997-08-20 | 卡尔·托马博士公司 | Novel sampling system for use in analysis of biological processes |
| CN202246689U (en) * | 2011-06-24 | 2012-05-30 | 吴小禾 | Sampling and quenching equipment |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754337A (en) * | 2016-12-09 | 2017-05-31 | 大连工业大学 | A kind of cell metabolism quenching system and its cell metabolism quenching method |
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| Publication number | Publication date |
|---|---|
| CN102268366B (en) | 2013-04-17 |
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