CN204455072U - A kind of cell pretreatment device - Google Patents
A kind of cell pretreatment device Download PDFInfo
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- CN204455072U CN204455072U CN201420859900.7U CN201420859900U CN204455072U CN 204455072 U CN204455072 U CN 204455072U CN 201420859900 U CN201420859900 U CN 201420859900U CN 204455072 U CN204455072 U CN 204455072U
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Abstract
The utility model relates to biological plant manufacturing engineering field, disclose a kind of cell pretreatment device, this device comprises pending cell sample storage zone, damping fluid storage zone one, lysate storage zone, liquid delivery member one, liquid delivery member two, liquid delivery member three, cell sample treating pond, micro-filtration pond, liquid delivery member four, membrane sepn pond, small molecules collecting region.The utility model cell pretreatment device solves a difficult problem for cell sample process under space microgravity environment, ground and space environment can be applied to simultaneously, achieve the automatization preprocessing process of cell sample, obtain higher lysis efficiency and the macromole rate of recovery.
Description
Technical field
The utility model relates to biological plant manufacturing engineering field, specifically, relates to a kind of cell pretreatment device.
Background technology
In current bio-science research field, utilize space microgravity environment to carry out the research of the aspect such as analyzing and testing of cell cultures, biological sample separation and purification, various biological sample, not only there is great scientific research value, and there is commercial prospect widely.
Under space environment, the operation such as centrifugal, extraction, precipitation adopted cell treating processes is subject to microgravity impact and can not normally carries out.Therefore the new mode of design is needed to process biological samples such as cells, with separation and Extraction object product.Due to the severe condition under space microgravity environment, cell pretreatment and separation of biopolymer process very difficult, also lack in the prior art and a kind of under microgravity environment, pretreated device carried out to cell.
Utility model content
In order to overcome above-mentioned technical problem, the utility model provides a kind of cell pretreatment device, and structure is simple, improves lysis efficiency and the macromole rate of recovery.
To achieve these goals, the utility model provides a kind of cell pretreatment device, under being in microgravity environment, comprising:
Pending cell sample storage zone, damping fluid storage zone one, lysate storage zone, liquid delivery member one, liquid delivery member two, liquid delivery member three, cell sample treating pond, micro-filtration pond, liquid delivery member four, membrane sepn pond, small molecules collecting region;
Pending cell sample storage zone, damping fluid storage zone one, lysate storage zone connect the entrance of cell sample treating pond respectively by liquid delivery member one, liquid delivery member two, liquid delivery member three;
The outlet of cell sample treating pond connects the entrance in micro-filtration pond;
The outlet in micro-filtration pond is by the entrance of liquid delivery member four junctional membrane separate tank;
The outlet in described membrane sepn pond also connects the entrance of small molecules collecting region.
Further, this device also comprises:
Damping fluid storage zone two, liquid delivery member five, macromole collecting region;
Damping fluid storage zone two is by the outlet of liquid delivery member five junctional membrane separate tank, and the entrance in described membrane sepn pond also connects the entrance of macromole collecting region.
Further, described device also comprises waste liquid storage zone, liquid delivery member six;
The outlet of described cell sample treating pond connects waste liquid storage zone by liquid delivery member six.
Further, the aperture of the mould material of cell sample treating pond is 1-10 micron, and the aperture of the mould material in described micro-filtration pond is 0.1-0.45 micron, and the molecular weight cut-off MWCO of the mould material in described membrane sepn pond is for being not less than 5KD.
Further, liquid delivery member seven, liquid delivery member eight;
Wherein the outlet of macromole collecting region connects liquid delivery member seven, and the outlet of small molecules collecting region connects liquid delivery member eight.
Further, described cell sample treating pond, micro-filtration pond and membrane sepn pond comprise:
Pond body, in described pond, body interior has cavity, the entrance and exit that described cavity is offered with described pond body surface is respectively communicated with, and arranges mould material in described cavity, and described cavity is divided into the two portions being communicated with described entrance and described outlet by described mould material.
Further, described device also comprises cabinet, and described cabinet comprises box bottom, erects dividing plate and middle level dividing plate, and middle level dividing plate is horizontally fixed on the inwall of described cabinet, erects dividing plate and is vertically fixed on the inwall of described cabinet and contacts described box bottom;
Wherein, liquid delivery member one, liquid delivery member two, liquid delivery member three, liquid delivery member six, liquid delivery member four, liquid delivery member five, liquid delivery member seven and liquid delivery member eight are fixed on described box bottom; Cell sample treating pond, micro-filtration pond, membrane sepn pond, pending cell sample storage zone, damping fluid storage zone one, damping fluid storage zone two, lysate storage zone, waste liquid storage zone, macromole collecting region and small molecules collecting region are fixed on the dividing plate of middle level.
Cell pretreatment device described in the utility model, comprising: pending cell sample storage zone, damping fluid storage zone one, lysate storage zone, liquid delivery member one, liquid delivery member two, liquid delivery member three, cell sample treating pond, micro-filtration pond, liquid delivery member four, membrane sepn pond, macromole collecting region, small molecules collecting region; Pending cell sample storage zone, damping fluid storage zone one, lysate storage zone connect the entrance of cell sample treating pond respectively by liquid delivery member one, liquid delivery member two, liquid delivery member three; The outlet of cell sample treating pond connects the entrance in micro-filtration pond; The outlet in micro-filtration pond is by the entrance of liquid delivery member four junctional membrane separate tank; The outlet in described membrane sepn pond also connects the entrance of small molecules collecting region.By the scheme that the utility model provides, utilize liquid delivery member to manage throughout cleaning, the pre-treatment such as cracking and separating substances that cell sample is carried out in promotion in device and switching, this device can carry out the preprocessing process of cell sample under ground and microgravity environment, solve the difficulty of cell sample under space environment, achieve the automatization pre-treatment of cell sample, obtain higher lysis efficiency and the macromole rate of recovery.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of cell pretreatment device described in the utility model;
Fig. 2 is the structural representation in born of the same parents' sample preparation pond described in the utility model, micro-filtration pond, membrane sepn pond;
Fig. 3 is the structural representation of the casing of cell pretreatment device described in the utility model;
Fig. 4 is the structural representation of the expanding unit of cell pretreatment device described in the utility model;
Fig. 5 is the structural representation of the free stream cataphoresis chip in the utility model.
Embodiment
Below with reference to the accompanying drawings embodiment of the present utility model is described.The element described in an accompanying drawing of the present utility model or a kind of embodiment and feature can combine with the element shown in one or more other accompanying drawings or embodiment and feature.It should be noted that for purposes of clarity, accompanying drawing and eliminate expression and the description of parts that have nothing to do with the utility model, known to persons of ordinary skill in the art or process in illustrating.
The utility model embodiment provides a kind of cell pretreatment device, and this device comprises:
Pending cell sample storage zone 2, damping fluid storage zone 1, lysate storage zone 4, liquid delivery member 1, liquid delivery member 26, liquid delivery member 37, cell sample treating pond 8, micro-filtration pond 13, liquid delivery member 4 14, membrane sepn pond 15, small molecules collecting region 22.Pending cell sample quantity is about 10
5-10
7individual.
Wherein, pending cell sample storage zone 2, damping fluid storage zone 1, lysate storage zone 4 connect the entrance of cell sample treating pond 8 respectively by liquid delivery member 1, liquid delivery member 26, liquid delivery member 37.The outlet of cell sample treating pond 8 connects the entrance in micro-filtration pond 13.The outlet in micro-filtration pond 13 is by the entrance of liquid delivery member 4 14 junctional membrane separate tank 15.The outlet in described membrane sepn pond 15 also connects the entrance of small molecules collecting region 22.By this device, the cracking in cell sample treating pond 8 of pending enchylema, and the filtration through micro-filtration pond 13 and membrane sepn pond 15, the product after filtration enters small molecules collecting region 22.Liquid delivery member 1, liquid delivery member 26, liquid delivery member 37 and liquid delivery member 4 14 are in the apparatus for making liquid move to assigned direction.
Further, this device also comprises damping fluid storage zone 2 16, liquid delivery member 5 17, macromole collecting region 19.Damping fluid storage zone 2 16 is by the outlet of liquid delivery member 5 17 junctional membrane separate tank 15, and the entrance in described membrane sepn pond 15 also connects the entrance of macromole collecting region 19.By this device, the macromolecular substance that tunicle separate tank 15 is tackled is transported to macromole collecting region 19.
Further, this device also comprises waste liquid storage zone 11, liquid delivery member 6 10.The outlet of described cell sample treating pond 8 connects waste liquid storage zone 11 by liquid delivery member 6 10.
Below in conjunction with accompanying drawing, the cell pretreatment device that the utility model embodiment provides is described in detail.
As shown in Figure 1, the utility model embodiment provides a kind of cell pretreatment device, and this device comprises:
Pending cell sample storage zone 2, damping fluid storage zone 1, damping fluid storage zone 2 16, lysate storage zone 4, liquid delivery member 1, liquid delivery member 25, liquid delivery member 37, liquid delivery member 4 14, liquid delivery member 5 17, switch 12,18,25, cell sample treating pond 8, micro-filtration pond 13, membrane sepn pond, membrane sepn pond 15, macromole collecting region 19, small molecules collecting region 22.
Wherein, pending cell sample storage zone 2, damping fluid storage zone 1, lysate storage zone 4 connect the entrance of cell sample treating pond 8 respectively by liquid delivery member 1, liquid delivery member 25, liquid delivery member 37.The outlet of cell sample treating pond 8 connects the entrance in micro-filtration pond 13 by switch 12.The outlet in micro-filtration pond 13 is by the entrance of liquid delivery member 4 14 junctional membrane separate tank 15, and the entrance in membrane sepn pond 15 also connects the entrance of macromole collecting region 19 by switch 18.Damping fluid storage zone 2 16 passes through the outlet of liquid delivery member 5 17 and switch 25 junctional membrane separate tank 15 successively, and the outlet in described membrane sepn pond 15 also connects the entrance of small molecules collecting region 22.
In addition, this device also comprises switch 9, liquid delivery member 6 10 and waste liquid storage zone 11, and the outlet of cell sample treating pond 8 is also connected waste liquid storage zone 11 by switch 9 with liquid delivery member 6 10 successively.
Optionally, housing 1 is also comprised.
In the utility model embodiment, liquid delivery member is specifically as follows pump, such as volumetric pump, wing pump etc.Switch can be specifically valve, such as magnetic valve etc.
Further, this device also comprises switch 20 and 23, liquid delivery member 7 21 and liquid delivery member 8 24.
The wherein outlet of macromole collecting region 19 connecting valve 20 and liquid delivery member 7 21 successively, outlet connecting valve 23 and the liquid delivery member 8 24 successively of small molecules collecting region 22.
Wherein, the mould material of different pore size and material is provided with in cell sample treating pond 8, micro-filtration pond 13 and membrane sepn pond 15, for filtering.Such as, the aperture of the mould material of cell sample treating pond 8 is 1-10 micron, the aperture of the mould material in micro-filtration pond 13 is 0.1-0.45 micron, and the MWCO (Molecular Weight Cut Off, molecular weight cut-off) that the mould material in membrane sepn pond 15 can realize is not less than 5KD.Optionally, in the utility model embodiment, the aperture of the mould material of cell sample treating pond 8 is 5 microns, and the aperture of the mould material in micro-filtration pond 13 is 0.22 micron, and the MWCO that the mould material in membrane sepn pond 15 can realize is 5KD.It is pointed out that above-mentioned all mould materials can be changed according to different sample and separation object in addition.
Further, the structure in cell sample treating pond 8, micro-filtration pond 13 and membrane sepn pond 15 all can see Fig. 2, for cell sample treating pond 8, wherein have cavity 805 in pond body 802 inside, the entrance 801 offered with pond body 802 surface respectively of cavity 805 with export 804 and be communicated with, arrange mould material 803 in cavity 805, cavity 805 is divided into two portions by mould material 803.Wherein a part is communicated with entrance 801, and another part is communicated with outlet 804.
Further, this device also comprises cabinet, and as shown in Figure 3, cabinet comprises box bottom 101, erects dividing plate 102 and middle level dividing plate 103.Middle level dividing plate 103 is horizontally fixed on the inwall of cabinet by screw, erects dividing plate 102 and is vertically fixed on the inwall of described cabinet and contacts described box bottom 101.Visible in figure 3, middle level dividing plate 103 and the vertical setting mutually of setting dividing plate 102, be divided into three parts by whole cabinet.
Wherein, liquid delivery member 1, liquid delivery member 25, liquid delivery member 37, liquid delivery member 6 10, liquid delivery member 4 14, liquid delivery member 5 17 is fixed on (can be specifically fixed thereon by the threaded hole on box bottom 101) on box bottom 101, switch 9, 12, 18, 25 are fixed on (can be specifically in the embedded hole slot being fixed on setting dividing plate 102) on setting dividing plate 102, cell sample treating pond 8, micro-filtration pond 13, membrane sepn pond 15, pending cell sample storage zone 2, damping fluid storage zone 1, damping fluid storage zone 2 16, lysate storage zone 4, waste liquid storage zone 11, macromole collecting region 19 and small molecules collecting region 22 are fixed on middle level dividing plate 103 (concrete, can be fixed thereon by the threaded hole of middle level dividing plate 103).The each several part of cell pretreatment device is provided separately, and is conducive to operation and maintenance.
For aforementioned cells pretreatment unit, the utility model embodiment still provides a kind of cell pretreatment method, and the method comprises:
S1, open liquid delivery member 1 cell suspension of pending cell sample storage zone 2 is filtered by cell sample treating pond 8, make cell sample be trapped in the mould material surface of described cell sample treating pond 8.
Concrete, filtered by the cell sample treating pond 8 that filter sizes is 5 microns by 1 ml cells suspension, nutrient solution and impurity enter waste liquid storage zone 11 at filter membrane surface to make cell retention, thus achieve being separated of cell and nutrient solution.
S2, open liquid delivery member 25 cell sample of the damping fluid of damping fluid storage zone 1 to the mould material surface of cell sample treating pond 8 is cleaned.
Wherein, the damping fluid in S2 is the PBS (phosphate buffer saline, phosphate buffered saline buffer) of precooling.By this damping fluid, cell sample is cleaned, wash away the medium component that cell surface is residual.
S3, open liquid delivery member 37 and make the cell sample of the lysate of lysate storage zone 4 to the mould material surface of described cell sample treating pond 8 carry out cracking.
The lysate system proportioning of lysate storage zone 4 is 1 milliliter of RIPA (RadioImmunoprecipitation Assay, radioimmunoprecipitation) lysate adds 10 microlitre PMSF (phenylmethylsulfonyl fluoride), i.e. 1ml RIPA+10 μ l PMSF.Get 1 milliliter of this lysate system to enter in cell sample treating pond 8 through liquid delivery member 37.The cleaved immersion profit of cell on film surface, and fully contacts with cracking system.
S4, change the electrode of liquid delivery member 37, make liquid delivery member 37 carry out forward and reverse switching according to predetermined frequency, the cell sample on the mould material surface of described cell sample treating pond 8 is vibrated filtration repeatedly.
Liquid delivery member 37 can two-way circulate, by changing liquid delivery member 37 electrode in S4, make the flow direction of liquid delivery member 37 carry out forward and reverse switching according to preset frequency, cell sample is vibrated filtration repeatedly, abundant cracking, improves the lysis efficiency of cell.
S5, open liquid delivery member 4 14, switch 12, by repeatedly vibrating, the cracking system after filtering is filtered through micro-filtration pond 13, and 15 pairs of macromolecular substance retain through membrane sepn pond, the small-molecule substance through membrane sepn pond 15 is sent into small molecules collecting region 22.
After opening liquid delivery member 4 14 and switch 12, cracking system enters micro-filtration pond 13 and filters.Optionally, the membrane pore size of the mould material in micro-filtration pond 13 is 0.22 micron.By filtering the impurity such as the uncracked cell debris of elimination, ensureing extract purity and preventing line clogging.Afterwards, in membrane sepn pond 15, biomacromolecule albumen is trapped within film.Optionally, the MWCO of this mould material is not less than 5KD.
S6, open liquid delivery member 5 17, switch 25, switch 18, the macromolecular substance that membrane sepn pond 15 is retained by the damping fluid of damping fluid storage zone 2 16 sends into macromole collecting region 19.
Macromolecular substance is rinsed repeatedly, and is collected in macromole collecting region 19.
In the utility model embodiment, be 20-40 minute (being preferably 30 minutes) to the pyrolysis time of the cell sample on the mould material surface of cell sample treating pond (8).The electrode of liquid delivery member (7) was every 3-5 minute (being preferably 3 minutes) conversion direction, run 10-15 second (being preferably 10 seconds) after each conversion direction, flow velocity is 1-3 ml/min (being preferably 1 ml/min).
The cell pretreatment device that the utility model embodiment scheme provides, cell sample treating pond is set, micro-filtration pond, membrane sepn pond carry out bed filtration extraction to cell suspension, respectively macromolecular substance and small-molecule substance are collected macromole collecting region and small molecules collecting region, and use liquid delivery member (such as pump) and switch (such as valve) control solution flowing and flow to, and positive and negative the transferring of the electrode controlling liquid delivery member improves cell sample lysis efficiency.This device integral construction is simple, is suitable for carrying out cell sample pre-treatment under microgravity environment, achieves the automation process of cell sample cleaning, cracking, macromole and separation of small molecuies, obtains higher lysis efficiency and the macromole rate of recovery.
Further, as shown in Figure 4, this device also comprises damping fluid storage zone 3 26, free stream cataphoresis chip 27, albumen storage zone 28.Macromole collecting region 19 is connected to free stream cataphoresis chip 27 through switch 20 and liquid delivery member 21.Damping fluid storage zone 3 26 is connected to free stream cataphoresis chip 27 through switch 29 and liquid delivery member 30.Free stream cataphoresis chip 27 connects albumen storage zone 28, and connects nucleic acid storage zone 31.Liquid delivery member 32,33 is also connected with, for liquid-flow provides power to supplement in device.
The electrophoresis path both sides of free stream cataphoresis chip 27 arrange electrode, and the positive and negative electrode of power supply (not shown) is connected in two lateral electrodes.The structural representation of free stream cataphoresis chip 27 is with reference to figure 5.
Free stream cataphoresis chip 27 inside is provided with electrophoresis path 2707.The both sides of electrophoresis path 2707 are electrode 2706, and the positive and negative electrode of power supply is added in two lateral electrodes 2706 respectively.The inflow end of electrophoresis path arranges sample inlet 2701, flows into end and is also provided with buffer inlet 2702,2703,2704 and 2705.Wherein 2702 and 2703 be connected to damping fluid storage zone 3 26, the damping fluid of damping fluid storage zone 3 26 injects buffer inlet 2702 and 2703.2704 are not connected damping fluid storage zone with 2705.If a newly-increased damping fluid storage zone, then can be connected to 2704 and 2705.
In practical application, the damping fluid storage zone be connected with free stream cataphoresis chip 27 can be more than one.The quantity of buffer inlet is not defined as four yet.Such as, only have a buffer inlet at inflow end, damping fluid storage zone 3 26 connects this buffer inlet and injects damping fluid.Or have two buffer inlet at inflow end, each buffer inlet connects a damping fluid storage zone.Or the buffer inlet of more than four also can be set.
At the outflow end of free stream cataphoresis chip 27, albumen outlet 2708, waste liquid outlet 2709 and nucleic acid outlet 2710 are set.Albumen outlet 2708 and nucleic acid outlet 2710 are arranged on the position near electrode.Waste liquid outlet 2709 is arranged on albumen outlet 2708 and nucleic acid exports between 2710, macromolecular sample is under electric field action, be separated fully in the electrophoresis path of free stream cataphoresis chip 27, protein molecule flows out from albumen outlet 2708 and enters albumen storage zone 28, nucleic acid molecule flows out from nucleic acid outlet 2710, enters nucleic acid storage zone 29.Neutral liquid flows out from waste liquid outlet 2709, and enters waste liquid storage zone 11 through switch 34 and liquid delivery member 10.
In this device as shown in Figure 4, also comprise moving phase storage zone 35, chromatographic column 36, temperature-controlled member 37, RNA collecting region 38, DNA collecting region 39.The outlet of chromatographic column 36 connects DNA collecting region 39 through switch 40 and connects RNA collecting region 38 through switch 41.Nucleic acid storage zone 31 is connected the entrance of chromatographic column 36 by liquid delivery member 42 and switch 43, and moving phase storage zone 35 is connected the entrance of chromatographic column 36 by liquid delivery member 44 and switch 45.Waste liquid storage zone 11 connects the outlet of chromatographic column 36 by liquid delivery member 10.Temperature-controlled member 37 is wrapped in chromatographic column 36.
RNA collecting region 38 the other end is the other end of connecting valve 46 and liquid delivery member 47, DNA collecting region 39 and connecting valve 48 and liquid delivery member 49 also.Switch 43 goes back connecting valve 50 and liquid delivery member 51.
Various switches described in the utility model embodiment are used for opening and blocking liquid communication, and switch described in practical application can be valve member such as magnetic valve.Liquid delivery member described in the utility model embodiment is for promoting liquid-flow, and liquid delivery member described in practical application can be pump, such as volumetric pump wing pump etc.
The workflow of single unit system is described below with reference to Fig. 4.
First stage, obtain small molecules sample and macromolecular sample:
Open liquid delivery member 5 cell suspension of pending cell sample storage zone 2 is filtered by cell sample treating pond 8, make cell sample be trapped in the mould material surface of described cell sample treating pond 8.Concrete, filtered by the cell sample treating pond 8 that filter sizes is 5 microns by 1 ml cells suspension, nutrient solution and impurity enter waste collection district 11 at filter membrane surface to make cell retention, thus achieve being separated of cell and nutrient solution.
Opening liquid delivery member 6 makes the cell sample of the damping fluid of damping fluid storage zone 1 to the mould material surface of cell sample treating pond 8 clean.By this damping fluid, cell sample is cleaned, wash away the medium component that cell surface is residual.
Opening liquid delivery member 7 makes the cell sample of the lysate of lysate storage zone 4 to the mould material surface of described cell sample treating pond 8 carry out cracking.The cleaved immersion profit of cell on film surface, and fully contacts with cracking system.
Change the electrode of liquid delivery member 7, make liquid delivery member 7 carry out forward and reverse switching according to predetermined frequency, the cell sample on the mould material surface of cell sample treating pond 8 is vibrated filtration repeatedly.
By changing liquid delivery member 7 electrode, making the flow direction of liquid delivery member 7 carry out forward and reverse switching according to preset frequency, cell sample being vibrated filtration repeatedly, abundant cracking, improve the lysis efficiency of cell.
Open liquid delivery member 14, switch 12, by repeatedly vibrating, the cracking system after filtering is filtered through micro-filtration pond 13, and 15 pairs of macromolecular substance retain through membrane sepn pond, the small-molecule substance through membrane sepn pond 15 is sent into small molecules sample storage district 22.
After opening liquid delivery member 14 and switch 12, cracking system enters micro-filtration pond 13 and filters.Optionally, the membrane pore size of the mould material in micro-filtration pond 13 is 0.22 micron.By filtering the impurity such as the uncracked cell debris of elimination, ensureing extract purity and preventing line clogging.Afterwards, in membrane sepn pond 15, biomacromolecule albumen is trapped within film.Optionally, the MWCO of this mould material is not less than 5KD.
Open liquid delivery member 17, switch 25, switch 18, the macromolecular substance that membrane sepn pond 15 is retained by the damping fluid of damping fluid storage zone 2 16 sends into macromolecular sample storage zone 19.
Subordinate phase, obtains protein sample and nucleic acid samples:
Open switch 29 and liquid delivery member 30, the damping fluid of damping fluid storage zone 3 26 is injected free stream cataphoresis chip.Power supply is adjusted to predetermined voltage.This predetermined voltage scope at 100-600V, can be specifically 500V.
The macromolecular sample collected macromolecular sample storage zone 19 injects free stream cataphoresis chip 27, the protein in macromolecular sample and nucleic acid is separated under voltage effect and enters collecting protein district 28 and nucleic acid collecting region 31 respectively.Wherein, the flow velocity that macromolecular sample injects free stream cataphoresis chip 27 is 0.048-0.1 ml/min, specifically can select 0.08 ml/min.Due to the damping fluid being injected into certain pH value in free stream cataphoresis chip 27, macromolecular sample is injected into after free stream cataphoresis chip 27 due to positive and negative charges different on equipotential difference band, protein belt positive charge, nucleic acid belt negative charge, under electric field action, protein and nucleic acid start to be separated, and flow out respectively through albumen outlet 2708 and nucleic acid outlet 2710 (with reference to figure 5).By the injection speed of adjustment macromolecular sample and damping fluid and voltage swing, protein is regulated with being separated of nucleic acid.
Phase III, DNA isolation and RNA:
After nucleic acid substances is collected nucleic acid collecting region 31, closing switch 1 and switch 2 41, and the moving phase in moving phase storage zone 35 is injected chromatographic column 36.After closing switch 1 and switch 2 41, open switch 43, switch 45, and open liquid delivery member 44 and liquid delivery member 42, the moving phase in moving phase storage zone 35 is injected chromatographic column 36.In the process by moving phase balance chromatographic column 36, waste liquid flows into waste liquid storage zone 11.After this, control temperature-controlled member 37 and the temperature of chromatographic column 36 is down to the first preset temp.
Moving phase is acetonitrile solution or the methanol aqueous solution of 30%-90%, and those skilled in the art can select arbitrarily within the scope of this.
The nucleic acid samples of nucleic acid storage zone 31 is injected described chromatographic column 36.Concrete, temperature reaches the first preset temp, and (scope of the first preset temp is 8-12 degree Celsius, preferably 10 degrees Celsius) after, close the liquid delivery member 10 connecting waste collection district 11, closing switch 45, opens liquid delivery member 42 and makes nucleic acid samples enter chromatographic column 9 with the flow velocity of 0.1 ml/min.Opening switch 50 and liquid delivery member 51 can carry out cooperation adjustment to flow velocity.Be filled with boric acid chromatographic material stationary phase in chromatographic column, these materials belong to temperature sensing material, the RNA sample in nucleic acid samples are caught at 10 degrees centigrade, thus make DNA sample flow out chromatographic column 36.
Opening switch 1, is collected into DNA collecting region 39 by DNA sample.
Closing switch 1, controls temperature-controlled member 37 and the temperature of chromatographic column 36 is risen to the second preset temp (scope of the second preset temp is 45-55 degree Celsius, is preferably 50 degrees Celsius).At this temperature, chromatographic column 36 loses the capture ability to RNA sample, and RNA sample flows out chromatographic column 36.Opening switch 2 41, is collected into RNA collecting region 38 by RNA sample.
By above-mentioned steps flow process, small-molecule substance is collected into small molecules sample storage district 22, and protein molecular is collected into collecting protein district 28, DNA and RNA collects DNA collecting region 39 and RNA collecting region 38 respectively.The device that the utility model embodiment provides achieve to the close-coupled processing of biological sample be separated, obtain four kinds of separate substances.This apparatus structure is clear, principle is simple, collection cell pretreatment, macromole separation of small molecuies, nucleic acid protein are separated, DNA and RNA is located away from all over the body, there is higher processing efficiency, high to macromolecular substance cracking organic efficiency, without the need to too much manpower intervention, be namely applicable to run under traditional gravity environment, also can be used for space microgravity environment.And automatic operating degree is high, be convenient to the application of extensive integrated industrial.
Although described the utility model and advantage thereof in detail, be to be understood that and can have carried out various change when not exceeding the spirit and scope of the present utility model limited by appended claim, substituting and conversion.And the scope of the application is not limited only to the specific embodiment of process, equipment, means, method and step described by specification sheets.One of ordinary skilled in the art will readily appreciate that from disclosure of the present utility model, can use perform the function substantially identical with corresponding embodiment described herein or obtain and its substantially identical result, existing and that will be developed in the future process, equipment, means, method or step according to the utility model.Therefore, appended claim is intended to comprise such process, equipment, means, method or step in their scope.
Claims (7)
1. a cell pretreatment device, is characterized in that, under described device is in microgravity environment, comprising:
Pending cell sample storage zone (2), damping fluid storage zone one (3), lysate storage zone (4), liquid delivery member one (5), liquid delivery member two (6), liquid delivery member three (7), cell sample treating pond (8), micro-filtration pond (13), liquid delivery member four (14), membrane sepn pond (15), small molecules collecting region (22);
Pending cell sample storage zone (2), damping fluid storage zone one (3), lysate storage zone (4) respectively by liquid delivery member one (5), liquid delivery member two (6), liquid delivery member three (7) connect cell sample treating pond (8) entrance;
The outlet of cell sample treating pond (8) connects the entrance in micro-filtration pond (13);
The outlet in micro-filtration pond (13) is by the entrance of liquid delivery member four (14) junctional membrane separate tank (15);
The outlet of described membrane sepn pond (15) also connects the entrance of small molecules collecting region (22).
2. cell pretreatment device according to claim 1, is characterized in that, also comprise:
Damping fluid storage zone two (16), liquid delivery member five (17), macromole collecting region (19);
Damping fluid storage zone two (16) is by the outlet of liquid delivery member five (17) junctional membrane separate tank (15), and the entrance of described membrane sepn pond (15) also connects the entrance of macromole collecting region (19).
3. cell pretreatment device according to claim 2, is characterized in that, also comprise:
Waste liquid storage zone (11), liquid delivery member six (10);
The outlet of described cell sample treating pond (8) connects waste liquid storage zone (11) by liquid delivery member six (10).
4. cell pretreatment device according to claim 3, it is characterized in that, cell sample treating pond is provided with mould material in (8), the aperture of the mould material of cell sample treating pond (8) is 1-10 micron, the aperture of the mould material of described micro-filtration pond (13) is 0.1-0.45 micron, and the molecular weight cut-off of the mould material of described membrane sepn pond (15) is for being not less than 5KD.
5. cell pretreatment device according to claim 3, is characterized in that, also comprise:
Liquid delivery member seven (21), liquid delivery member eight (24);
Wherein the outlet of macromole collecting region (19) connects liquid delivery member seven (21), and the outlet of small molecules collecting region (22) connects liquid delivery member eight (24).
6. cell pretreatment device according to claim 3, is characterized in that, described cell sample treating pond (8), micro-filtration pond (13) and membrane sepn pond (15) comprise respectively:
Pond body, in described pond, body interior has cavity, the entrance and exit that described cavity is offered with described pond body surface is respectively communicated with, and arranges mould material in described cavity, and described cavity is divided into the two portions being communicated with described entrance and described outlet by described mould material.
7. according to the cell pretreatment device in claim 3 to 6 described in any one, it is characterized in that, described device also comprises cabinet, described cabinet comprises box bottom (101), erects dividing plate (102) and middle level dividing plate (103), middle level dividing plate (103) is horizontally fixed on the inwall of described cabinet, erects dividing plate (102) and is vertically fixed on the inwall of described cabinet and contacts described box bottom (101);
Wherein, liquid delivery member one (5), liquid delivery member two (6), liquid delivery member three (7), liquid delivery member six (10), liquid delivery member four (14), liquid delivery member five (17), liquid delivery member seven (21) and liquid delivery member eight (24) are fixed on described box bottom (101); Cell sample treating pond (8), micro-filtration pond (13), membrane sepn pond (15), pending cell sample storage zone (2), damping fluid storage zone one (3), damping fluid storage zone two (16), lysate storage zone (4), waste liquid storage zone (11), macromole collecting region (19) and small molecules collecting region (22) are fixed on middle level dividing plate (103).
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| CN201420859900.7U CN204455072U (en) | 2014-12-30 | 2014-12-30 | A kind of cell pretreatment device |
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| CN201420859900.7U CN204455072U (en) | 2014-12-30 | 2014-12-30 | A kind of cell pretreatment device |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560636A (en) * | 2014-12-30 | 2015-04-29 | 北京理工大学 | Cell pretreatment device and method |
| CN109456882A (en) * | 2019-01-16 | 2019-03-12 | 北京理工大学 | A kind of space biological sample micro process integrating device |
| CN110198785A (en) * | 2016-11-08 | 2019-09-03 | 佐治亚科技研究公司 | Method for convection current driving intracellular delivery |
-
2014
- 2014-12-30 CN CN201420859900.7U patent/CN204455072U/en active Active
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104560636A (en) * | 2014-12-30 | 2015-04-29 | 北京理工大学 | Cell pretreatment device and method |
| CN110198785A (en) * | 2016-11-08 | 2019-09-03 | 佐治亚科技研究公司 | Method for convection current driving intracellular delivery |
| CN109456882A (en) * | 2019-01-16 | 2019-03-12 | 北京理工大学 | A kind of space biological sample micro process integrating device |
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