CN209485831U - The extracellular vesica enriching apparatus of the urine of hospital - Google Patents
The extracellular vesica enriching apparatus of the urine of hospital Download PDFInfo
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- CN209485831U CN209485831U CN201821324225.2U CN201821324225U CN209485831U CN 209485831 U CN209485831 U CN 209485831U CN 201821324225 U CN201821324225 U CN 201821324225U CN 209485831 U CN209485831 U CN 209485831U
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Abstract
Utility model device is suitable for field of medical technology, is a kind of extracellular vesica enriching apparatus of the urine that hospital uses, and can be used for the non-invasive liquid biopsy of chronic disease early diagnosis.The device includes: filter, it is set to the upper end of described device, including solution cavity and filter layer, after the solution cavity is added in the supernatant of urine specimen, retention is greater than the extracellular vesica in the aperture of the filter layer, and filtration is less than other urinary fractions in the extracellular vesica aperture；Liquid waste collector has the collecting chamber for accommodating other urinary fractions less than the extracellular vesica aperture in the urine, and the liquid waste collector is socketed on the outside of the filter；Vacuum suction interface is provided on the liquid waste collector side wall；Negative pressure component is connected to the vacuum suction interface, will be aspirated between the liquid waste collector and the filter as preset negative pressure.Whereby, the utility model realizes the convenient extraction of the extracellular vesica of urine.
Description
Technical field
Utility model device is related to the field of medical instrument technology more particularly to a kind of extracellular vesica of the urine of hospital
Enriching apparatus can be used for the non-invasive liquid biopsy of chronic disease early diagnosis.
Background technique
Urine is as a kind of most common detection various disease specific biomarker object, for diagnosing the sample of various disease
Source, materials are convenient, and the invasive inspection method such as blood preparation can be overcome to be not easy to be accepted by patients, is more difficult by general population
The shortcomings that human health screening is received.But there is a large amount of physiological protein (such as Tamm-Hosfall albumen) high abundance in urine
The interference of ingredient makes us be not easy to detect low abundance but the urinary fractions of the reaction disease variation degree that has diagnostic value.
The ingredients such as albumen, lipid, nucleic acid in the extracellular vesica of urine EVs (Extracelluar Vesicles, EVs), although can make
For identify various disease biomarker, but mostly be also low abundance ingredient pass through there is an urgent need to research and develop a kind of enriching apparatus
It is enriched with EVs, to significantly improve the recall rate of the low abundance ingredient of EVs.The utility model belongs to biotechnology and clinical examination skill
Art field more particularly to a kind of nanometer film enrichment extracellular vesica of urine for hospital application are used to early diagnose a variety of chronic
The device of disease.
Extracellular vesica is that a kind of endocytosis system that originates from is united, secreted by way of plasma membrane fusion by cell multivesicular body
Film property vesica, EVs includes there are many protein, lipid, various RNA (ribonucleic acid, ribonucleic acid) ingredient etc.,
It can be secreted, be widely distributed in various body fluid by a variety of different types of cells, intercellular carrier can be used as, pass through cargo
Function vector transmits various physiology and pathological information, has and participates in intracellular signal transduction, immunological regulation, establishes tumor escape machine
System mediates the functions such as regeneration and degenerative process, is the novel targets with clinical diagnosis and the treatment of high development potentiality, different diseases
The secretory volume of extracellular vesica and its component content such as protein, lipid, RNA etc. can produce variation when sick, these variations were both
Related with the pathogenesis of various disease, the difference of the secretory volume and component content that detect extracellular vesica can be used for different diseases
The antidiastole of disease.
Urine is as a kind of most common detection various disease specific biomarker object, for diagnosing the sample of various disease
Source, materials are convenient, and the invasive inspection method such as blood preparation can be overcome to be not easy to be accepted by patients, is more difficult by general population
The shortcomings that human health screening is received.
Outside existing enrichment of cell in the technical solution of vesica, including following several:
(1) supercentrifugation carries out the separation of sample EVs using different ultracentrifugation speed or density gradient centrifugation
And extract, it is current EVs separating and extracting process only applied in scientific research institutions, expensive.The obtained EVs of supercentrifugation
Although with high purity, due to needing equipped with expensive Ultracentrifuge and corresponding consumptive material, and entire extraction process consumption
Shi Manchang is only applicable to the processing of a small amount of sample.
(2) it by the magnetic bead absorption method of immunoaffinity capture, is incubated together with sample using the magnetic bead containing specific antibodies
Reaction is educated, separates magnetic bead-EVs association in conjunction with the EVs that film surface in sample contains corresponding antigen, then through magnetic-adsorption
Out.Magnetic bead absorption method separates specific EVs purity is high although extracting, and can distinguish hypotype, but needs to carry out special processing to magnetic bead
Processing, reagent consumptive material consumption is big, and productivity and yield is low.
(3) the kit extracting method based on co-precipitation, the commercial kit of Exo-Quick, Exo-Spin etc.
It is co-precipitated principle using polymerization, the aquation that reagent inhibits EVs is added in sample, makes it easier to precipitate, then lead to
Later continue low-speed centrifugal to be enriched with to obtain the precipitating of EVs.The shortcomings that kit extracting method based on co-precipitation, is to generate poly-
The impurity such as particle, lipoprotein, RNA compound are closed, resulting EVs purity is lower, and kit is expensive, can not be extensive
It applies in clinic.
In summary, the prior art in actual use, require professional technician that general hospital does not have and
Professional equipment, cannot be in most of clinical applications, and it is therefore necessary to research and develop easy-to-use new technique device.
Utility model content
For above-mentioned defect, the purpose of this utility model is to provide a kind of extracellular vesica enrichments of the urine of hospital
Device, it is thus achieved that the convenient extraction of the extracellular vesica of urine.
To achieve the goals above, the utility model provides a kind of extracellular vesica enriching apparatus of urine of hospital, packet
It includes:
Filter is set to the upper end of described device, including solution cavity and filter layer, and the supernatant of urine specimen is added
After the solution cavity, retention is greater than the extracellular vesica in the aperture of the filter layer, and filtration is less than the extracellular vesica aperture
Other urinary fractions；
Liquid waste collector has and accommodates other urinary fractions less than the extracellular vesica aperture in the urine
Collecting chamber, and the liquid waste collector is socketed on the outside of the filter；It is provided on the liquid waste collector side wall true
Suction gas interface；
Negative pressure component is connected to the vacuum suction interface, will aspirate between the liquid waste collector and the filter
For preset negative pressure.
According to the extracellular vesica enriching apparatus of the urine of the hospital, the filter layer includes:
Nanometer film support frame, the nanometer film support frame are set to the bottom of the filter, the side with the filter
Wall connection；
Nanofiltration membrane is set to the inside of the nanometer film support frame.
According to the extracellular vesica enriching apparatus of the urine of the hospital, the nanometer film support frame includes multiple first
One end of bracket item, multiple first support items is connect with the bottom of the side wall of the filter respectively, and the other end is connected to one
It rises；The nanofiltration membrane has multiple first filter hole, and the diameter of multiple first filter hole is 20 nanometers.
According to the extracellular vesica enriching apparatus of the urine of the hospital, the nanofiltration membrane is that can filter volume to be less than
The soluble protein of extracellular vesica and the nanofiltration membrane of other urinary fractions；The urine cell of the nanofiltration membrane retention
Outer vesica is collected in the solution cavity.
According to the extracellular vesica enriching apparatus of the urine of the hospital, the side wall upper part of the liquid waste collector is arranged
There is vacuum suction interface, the negative pressure component is vacuum pump；The vacuum suction interface is for connecting vacuum pump；
The top of described device has support edge, and the support is closing knot along the top nozzle with the liquid waste collector
Structure；
Described device further includes nut cap, is covered on the top of the sample pool, and the lateral wall with the liquid waste collector can
Dismantling connection is detachably connected with the support edge.
According to the extracellular vesica enriching apparatus of the urine of the hospital, the longitudinal cross-section of the nanofiltration layer is V
Font；
The shape of the filter and the liquid waste collector is in cylindrical；
The outer wall of the filter and the liquid waste collector is made of transparent material；
Graduation mark, and these described graduation marks are provided on the outer wall of the filter and the liquid waste collector
Not on same straight line in a lateral direction.
According to the extracellular vesica enriching apparatus of the urine of the hospital, described device further include:
Low speed centrifuge, including low speed turntable；
Sample shifts cup, and bottom is adapted with the low speed turntable；For holding urine specimen；
The low speed centrifuge centrifugation predetermined time removes the cell fragment in the urine specimen, obtains the supernatant
Liquid.
According to the extracellular vesica enriching apparatus of the urine of the hospital, the filter and the liquid waste collector can
Dismantling connection；And
The bottom of the liquid waste collector and the outer wall of the liquid waste collector are detachably connected.
According to the extracellular vesica enriching apparatus of the urine of the hospital, the top of the filter and the waste liquid are received
Compatible shackle member is provided at the top of storage；
The outer wall of the inner wall of the bottom of the liquid waste collector and the liquid waste collector is respectively arranged with corresponding inside and outside
Screw thread.
According to the extracellular vesica enriching apparatus of the urine of the hospital, described device further include:
For holding the washing cup of ultrapure water, it is set to that the liquid waste collector is external, the material of the washing cup is
Bright material, and graduation mark is provided on the washing cup, the capacity of the washing cup is 200 milliliters；
For blowing and beating the pipettor of the trapped fluid in the nanofiltration membrane, it is external to be set to the liquid waste collector；
For collecting the extracellular vesica collector of the extracellular vesica of the urine, it is set to the filter layer lower end, and
And extracellular vesica collector is removably connect with the nanometer film support frame of the filter layer；By the piping and druming of the pipettor
The extracellular vesica of urine is retained in the extracellular vesica collector.
The extracellular vesica enriching apparatus of the urine of the utility model hospital is set as including filter and liquid waste collector,
Filter is equipped with the support frame of nanometer film and lower section, and the top of filter is equipped with support edge, and support edge is with liquid waste collector top
Enclosed construction, filter lower half side wall are equipped with vacuum pump air-breathing interface tube, are controlled to a vacuum pump by air intake duct.It is first as a result,
Urine specimen is first removed into cell fragment by low speed centrifuge, then the supernatant of urine is put into filter, and passes through
Vacuum pump carries out negative pressure ventilation, is filtered using hydrostatic pressing and filter membrane pressure at both sides to urine specimen, and retention obtains EVs, and
Without complex operations, handled without other complex experiment instruments and reagent, it is simple and quick.The present apparatus uses consumables cost valence
Lattice are cheap, do not need special professional technician and expensive professional equipment, are suitble to promote hospital's large-scale use.
Detailed description of the invention
Fig. 1 is the extracellular vesica enriching apparatus structural schematic diagram of urine of hospital provided by the embodiment of the utility model；
Fig. 2 is the extracellular vesica enriching apparatus structural schematic diagram of urine of hospital provided by the embodiment of the utility model；
Fig. 3 is the extracellular vesica enriching apparatus structure diagram of urine of hospital shown in Fig. 2；
Nanometer film branch in the extracellular vesica enriching apparatus of the urine of Fig. 4 A hospital provided by the embodiment of the utility model
Strut configuration schematic diagram；
Nanometer film branch in the extracellular vesica enriching apparatus of the urine of Fig. 4 B hospital provided by the embodiment of the utility model
Strut configuration schematic diagram；
Fig. 5 is the extracellular vesica enriching apparatus enrichment urine cell of urine of hospital provided by the embodiment of the utility model
The inspection result schematic diagram of outer vesica.
Specific embodiment
In order to make the purpose of the utility model, technical solutions and advantages more clearly understood, below in conjunction with attached drawing and implementation
Example, the present invention will be further described in detail.It should be appreciated that specific embodiment described herein is only used to explain
The utility model is not used to limit the utility model.
Referring to FIG. 1 to FIG. 3, a kind of urine cell external capsule of hospital is provided in one embodiment of the utility model
Steep enriching apparatus 100, comprising:
Filter 10 is set to the upper end of device 100, including solution cavity 12 and filter layer 11；By the supernatant of urine specimen
After solution cavity 12 is added in liquid, retention is greater than the extracellular vesica in the aperture of the filter layer, and filtration is less than the extracellular vesica
Other urinary fractions in aperture；
Liquid waste collector 20 has other urinary fractions less than the extracellular vesica aperture accommodated in the urine
Collecting chamber 21, and liquid waste collector 20 is socketed on the outside of the filter 10；It is provided on 20 side wall of liquid waste collector
Vacuum suction interface 315；
Negative pressure component is connected to vacuum suction interface 315, is preset by aspirating between liquid waste collector 20 and filter
Negative pressure.
In this embodiment, the extracellular vesica enriching apparatus 100 of the urine of hospital includes that filter 10 and waste liquid are received
Storage 20.Wherein solution cavity 12 is the supernatant for placing urine specimen；Supernatant can be added portionwise.Filter 10 wraps
Filter layer 21 is included with filter membrane, the urine impurity in the supernatant can be filtered out, the extracellular vesica of urine is extracted, and
It is stored in solution cavity 12.Negative pressure component is connected to vacuum suction interface 315, will aspirate between liquid waste collector 20 and filter
For preset negative pressure, the efficiency being enriched with raising.The efficiency of the extracellular vesica enrichment of urine is improved by way of vacuumizing.It should
The extracellular vesica enriching apparatus 100 of the urine of hospital short, the extracellular vesica yield of gained low in cost, easy to operate, time-consuming
With with high purity and structural integrity, for the use of hospital's large-scale promotion.The extracellular vesica enriching apparatus 100 of the urine of hospital can
For most of clinical application applications, the device extracellular vesica obtained can be used for early diagnosing a variety of chronic diseases into one
Step tests and analyzes.
Referring to FIG. 1 to FIG. 3, in one embodiment of the utility model, filter layer 11 includes:
Filter layer 11, including nanometer film support frame 111 and nanofiltration membrane 113；Nanometer film support frame 111 is set to filter
10 bottom is connect with the side wall of filter 10；Nanofiltration membrane 113 is set on nanometer film support frame 111.The bottom of filter 10
Portion is that nanofiltration membrane 113, the nanofiltration membrane are embedded on the inside of nanometer film support frame 111 to the inclined nanometer film support frame 111 in middle part
113, in addition to embedded mode, can also be fixed by way of sticking.The central Formation cross-section of filter 10 is the circle of V-arrangement
The filter 10 of taper；Filter 10 and 20 outer wall of liquid waste collector are equipped with volume scale.
In addition, the side wall upper part of liquid waste collector 20 is provided with vacuum suction interface 315, vacuum suction interface 315 is used for
Connect vacuum pump；In addition, the top of device 100 has support along 32, and holds in the palm and be with the top nozzle of liquid waste collector 20 along 32
Enclosed construction.There is an enclosed construction at the edge of nanofiltration membrane 113, and 10 top nozzle of filter is equipped with support along 32, the support along 32 with
It is enclosed construction between 20 top nozzle of liquid waste collector, 20 top nozzle of liquid waste collector is equipped with nut cap, tube wall upper semisection outside
Vacuum suction interface 315 equipped with vacuum pump air intake duct, is docked by vacuum suction interface 315 with vacuum pump.This
Outside, filter 10 is detachably connected with liquid waste collector 20.The top of filter 10 and the top of liquid waste collector 20 are respectively provided with
There is compatible shackle member.The two removably connects, and can after use dismantle the two, be cleaned.For example, can be with
The lateral wall of liquid waste collector 20 is spirally connected, or is provided in the inside of nut cap 50 suitable along 32 phases with support with support along 32 clampings
The seal gasket matched.It is possible thereby to which the device 100 that the extracellular vesica of the urine of the hospital is extracted is closed, formation is vacuumized
Environment.When needing use device 100 to be enriched with the extracellular vesica of urine, after sample liquid is added, by nut cap 50 and waste collection
The lateral wall of device 20 screws, and after the completion of enrichment, then unscrews nut cap 50, carries out the cleaning etc. of device 100.
A and Fig. 4 B referring to fig. 4, nanometer film support frame 111 include multiple first support items 1111, multiple first support items
1111 one end is connect with the bottom of the side wall of filter 10 respectively, and the other end links together.Multiple first support items
1111 connect with the bottom of the side wall of filter 10, will constitute a pyramid type, if Fig. 4 A, such as Fig. 4 B are to be equipped with cell external capsule
Steep collector 25.Specifically, nanofiltration membrane 113 has multiple first filter hole, the diameter of multiple first filter hole is
20 nanometers.The diameter of the extracellular vesica of most of urine is usually 30 to 1000 nanometers.Therefore nanofiltration membrane 113 completely can be with
The effective extracellular vesica of urine extracted in supernatant.Preferably, which further includes, thin for collecting the urine
The extracellular vesica collector 25 of extracellular vesica is set to 11 lower end of filter layer, and extracellular vesica collector 25 and filtering
The nanometer film support frame 111 of layer 11 removably connects；The top of collector 25 is settable corresponding with nanometer film support frame 111
Flexible clamping and card hole, it is possible thereby to which extracellular vesica collector 25 is dismantled.By the piping and druming of the pipettor urine
Extracellular vesica is retained in the extracellular vesica collector 25；The side wall of liquid waste collector 20 and bottom 201 are detachably connected,
The two can be connected by shackle member, or the modes such as threaded connection.As nanometer film support frame 111 and extracellular vesica are collected
The top of device 25 be respectively arranged with relative to shackle member, after the side wall of liquid waste collector 20 and bottom 201 are dismantled
Extracellular vesica collector 25 is taken out.In one embodiment of the utility model, device 100 further include:
Low speed centrifuge, including low speed turntable；
Sample shifts cup, and bottom is adapted with the low speed turntable；For holding urine specimen；
The low speed centrifuge rotation predetermined time removes the cell fragment in the urine specimen, obtains the supernatant
Liquid.Urine specimen is placed in sample transfer cup, is then placed it on low speed centrifuge, it can will be thin by low speed centrifuge
The removal of born of the same parents' fragment urine impurity；Then supernatant is poured into filter 10 by sample transfer cup.Sample transfer cup has glass appearance
Device is made, and has graduation mark.Due to 113 layers of the nanofiltration membrane filtering urines impurity such as including soluble protein；By nanofiltration membrane
The extracellular vesica of 113 layers of the retention urine is collected in solution cavity 12.
Furthermore described device 100 further include: for holding the washing cup of ultrapure water, be set to 20 body of liquid waste collector
Outside, the material of the washing cup is transparent material, and graduation mark is provided on the washing cup, and the capacity of the washing cup is
200 milliliters；For blowing and beating the pipettor of the trapped fluid in the nanofiltration membrane 113, it is external to be set to the liquid waste collector 20；
And connect with the vacuum suction interface 315, the vacuum pump for vacuumizing.In addition, the enrichment extracellular vesica of urine
Device 100 further includes nut cap 50, is covered on the top of sample pool 10, is spirally connected with the lateral wall of liquid waste collector 20.It is possible thereby to
The device 100 for being enriched with the extracellular vesica of urine is closed, form the good environment vacuumized.It is thin using the enrichment urine when needing
When the device 100 of extracellular vesica is enriched with urine extracellular vesica, after sample liquid is added, by the outer of nut cap 50 and liquid waste collector 20
Side wall screws, and after the completion of enrichment, then unscrews nut cap 50, and the taking-up of the progress extracellular vesica of urine, or enrichment urine are thin
The cleaning of the device 100 of extracellular vesica.It is respectively arranged on the inside of nut cap 50 and the lateral wall of liquid waste collector 20
Corresponding internal and external threads.Or again by extracellular vesica collector 25 after the side wall of liquid waste collector 20 and bottom 201 are dismantled
It dismantles, extracellular vesica collector 25 can be taken out from nanometer film support frame 111.
In one embodiment of the utility model, referring to fig. 4 the longitudinal cross-section of A and Fig. 4 B filter layer 11 be V-shape or
Person's circular arc type.The shape of filter 10 and liquid waste collector 20 is in cylindrical；Filter 10 and the waste collection
The outer wall of device 20 is made of transparent material.Quarter is provided on the outer wall of solution cavity 12, filter 10 and liquid waste collector 20
Line is spent, and on the same straight line of these described graduation marks not in a lateral direction.Thus, it is possible to be clearly seen that solution cavity
12, the capacity of the solution loaded in the scale on the outer wall of filter 10 and liquid waste collector 20, and current each container.
Referring also to Fig. 2 and Fig. 3, the bottom 201 of liquid waste collector and the outer wall of liquid waste collector 20 are detachably connected.The two can lead to
The mode for crossing the shackle member either threaded connection of adaptation is attached.The inner wall of the bottom 201 of specific liquid waste collector and
The outer wall of liquid waste collector 20 is respectively arranged with corresponding internal and external screw thread.It, can be by the way that waste liquid be received after the completion of enrichment process
The bottom 201 of storage is dismantled, and the work such as the cleaning of urine impurity collecting chamber 21 are carried out.
In one embodiment of the utility model, the extracellular vesica of the urine of hospital provided by the utility model is used
Enriching apparatus 100 extracts the extracellular vesica of urine, and the specific method is as follows:
Urine specimen is first put into low speed centrifuge in batches and is centrifuged, parameter of noncentricity 2000g, 30min, and room temperature is removed
Cell fragment etc. is removed, supernatant is collected.The sample handled is transferred in filter 10, vacuum pump air intake duct by transfer cup
Interface 315 connects upper vacuum pump and carries out negative pressure ventilation, carries out vesica enrichment using hydrostatic pressing and nanometer film pressure at both sides difference, passes through
EVs of the diameter at 30 to 1000 nanometers can be retained when nanofiltration membrane 113；The trapped fluid for finally collecting EVs solution cavity 12, that is, complete
The extraction of urine EVs.Preferably, it when trapped fluid residue 10ml or so in device 10 to be filtered, is enriched with washing cup toward EVs nanometers
200ml ultrapure water is added in device, continues the filtering process of above-mentioned nanofiltration membrane.When trapped fluid residue 5ml in device 10 to be filtered,
It is mixed with pipettor piping and druming trapped fluid and is collected into EVs collector 25, which is final urine cell external capsule
Steep solution.The utility model is low in cost, easy to operate, time-consuming short, gained EVs yield and purity is high and structural integrity, can be big
Scale extends to basic hospital use.
In one embodiment of the utility model, the extracellular vesica enrichment process of urine includes:
(1) low-speed centrifugal: taking the Urine specimens of 30ml healthy volunteer, studied, low speed centrifuge (AllegraTMX-
22Centrifuge, Beckman Coulter), relative centrifugal force (RCF) 2000g, room temperature, 30 minutes, removing cell fragment,
The sundries such as bacterium collect supernatant；
(2) nanofiltration membrane that pore size is 20 nanometers filters: collecting low-speed centrifugal treated supernatant, passes through transfer
Cup, is transferred in filter, and vacuum pump air-breathing interface tube connects upper vacuum pump and carries out negative pressure ventilation, utilizes hydrostatic pressing and nanometer film
Pressure at both sides difference carries out vesica enrichment, by that can retain EVs of the diameter at 30 to 1000 nanometers when nanofiltration membrane；
(3) it the collection of extracellular vesica: when trapped fluid residue 10ml in device to be filtered or so, is received with washing cup toward EVs
200ml ultrapure water is added in rice enricher, continues the filtering process of above-mentioned nanofiltration membrane.Trapped fluid in device to be filtered is remaining
When 5ml, it is mixed with pipettor piping and druming trapped fluid and is collected into EVs collector, which is final urine cell
Outer vesicle solution.
The nanofiltration membrane 113 but using purchased from bodyguard must pure (Shanghai) trade Co., Ltd Spectrum Labs Inc, hole
The nanofiltration membrane that diameter size is 20 nanometers retains EVs of the diameter at 30 to 1000 nanometers.
(4) transmission electron microscope observing: taking the extracellular vesica 20ul of urine prepared in step (3), and point sample is in copper mesh, and 5 minutes
Afterwards, 3% (W/V) phosphotungstic acid negative staining is given 2 minutes, twice of milli-Q water, drying detects after five minutes, transmission electron microscope focal length is adjusted,
The form and partial size for observing urine excretion body, as shown in concrete outcome Fig. 5, from fig. 5, it can be seen that the extracellular vesica of urine point
Cloth is uniform, and cup-shaped or round, partial size is in 40-1000nm.
(5) NanoSight is detected: using the extracellular vesica of PBS diluted urine, proportional diluted is diluted to 200 times, on syringe
Sample, each 2-3ml, loading front and back clean detection slot with ultrapure water, NanoSight instrument parameter are adjusted, until proper focal length, right
Extracellular vesica carries out analysis record.For the partial size main peak of the extracellular vesica of testing result urine in 102nm, sample is purer.
(6) testing result
The extracellular vesica of urine is cup-shaped or spherical under transmission electron microscope, is evenly distributed, shown in specific visible Fig. 5.
The mean particle size of the extracellular vesica of urine is 102nm.
The extracellular vesica of urine is evenly distributed, and Brownian movement is significant.
It is provided by the utility model compared with existing common supercentrifugation, magnetic bead absorption method and isolation kit method
Having the advantage that for extracellular vesica carries out the extracellular vesica of urine using the nanometer film of special pore size distribution and molecular cut off
Concentration and separation, the preferably nanometer film of 20nm in the present embodiment, molecular cut off 1000kD.Urine is pre-processed by centrifugation,
It can remove the cell fragment of the overwhelming majority.Acquisition and the comparable extracellular vesica of above method purity can be enriched with.Because of the present apparatus
It can retain EVs of the diameter at 30 to 1000 nanometers when aperture is 20 nanometers of nanofiltration membrane 113 in 100, and remaining soluble egg
The urine impurity etc. that white equal diameter is less than extracellular vesica is then filtered out from nanofiltration membrane 113.The present apparatus 100 can be handled disposably greatly
Urine specimen is measured, and is not necessarily to complex operations.The present apparatus 100 is configured with minipump and low speed centrifuge, complicated without other
Laboratory apparatus and reagent processing.The present apparatus 100 carries out negative pressure ventilation by vacuum pump, poor using hydrostatic pressing and filter membrane pressure at both sides
It realizes, it is simple and quick.The present apparatus 100 is easy to carry, cheap using consumables cost, is suitble to push away in most of hospitals
It is wide to use.
The utility model is by including filter and waste collection for hospital's extracellular vesica enriching apparatus setting of urine
Device, filter are equipped with nanometer filtration membrane, and filter interior forms EVs collector, inside liquid waste collector and filter lower section
Between form the waste collection chamber of urine impurity, the tip edge of filter is equipped with support edge, and the support along being with liquid waste collector top
Enclosed construction, liquid waste collector tube wall are equipped with vacuum pump air-breathing interface tube, are controlled to a vacuum pump by air intake duct.Thus, it is possible to
Urine specimen is removed into cell fragment by low speed centrifuge first, then urine supernatant is put into filter, and pass through vacuum
Pump carries out negative pressure ventilation, is filtered using hydrostatic pressing and filter membrane pressure at both sides to urine specimen, obtains EVs, and without complexity
Operation is handled without other complex experiment instruments and reagent, simple and quick.The present apparatus is cheap using consumables cost,
Special professional technician and expensive professional equipment are not needed, is suitble to extend to hospital's large-scale use.
Certainly, the utility model can also have other various embodiments, without departing substantially from the spirit of the present invention and its essence
In the case of, those skilled in the art work as can make various corresponding changes and modifications, but these according to the utility model
Corresponding changes and modifications all should belong to the protection scope of the utility model the attached claims.
Claims (10)
1. a kind of extracellular vesica enriching apparatus of the urine of hospital characterized by comprising
Filter is set to the upper end of described device, including solution cavity and filter layer, described in the supernatant addition by urine specimen
After solution cavity, retention is greater than the extracellular vesica in the aperture of the filter layer, and filtration is less than its of the extracellular vesica aperture
His urinary fractions；
Liquid waste collector has the collection for accommodating other urinary fractions less than the extracellular vesica aperture in the urine
Chamber, and the liquid waste collector is socketed on the outside of the filter；Vacuum suction is provided on the liquid waste collector side wall
Gas interface；
Negative pressure component is connected to the vacuum suction interface, is pre- by suction between the liquid waste collector and the filter
If negative pressure.
2. the extracellular vesica enriching apparatus of the urine of hospital according to claim 1, which is characterized in that the filter layer
Include:
Nanometer film support frame, the nanometer film support frame are set to the bottom of the filter, connect with the side wall of the filter
It connects；
Nanofiltration membrane is set to the inside of the nanometer film support frame.
3. the extracellular vesica enriching apparatus of the urine of hospital according to claim 2, which is characterized in that the nanometer film
Support frame includes multiple first support items, and one end of multiple first support items connects with the bottom of the side wall of the filter respectively
It connects, the other end links together；The nanofiltration membrane has multiple first filter hole, the diameter of multiple first filter hole
It is 20 nanometers.
4. the extracellular vesica enriching apparatus of the urine of hospital according to claim 3, which is characterized in that the nanometer filter
Film is that can filter volume less than the soluble protein of extracellular vesica and the nanofiltration membrane of other urinary fractions；The nanofiltration membrane
The extracellular vesica of the urine of retention is collected in the solution cavity.
5. the extracellular vesica enriching apparatus of the urine of hospital according to claim 2, which is characterized in that the waste liquid is received
The side wall upper part of storage is provided with vacuum suction interface, and the negative pressure component is vacuum pump；The vacuum suction interface is for connecting
Connect vacuum pump；
The top of described device has support edge, and the support is enclosed construction along the top nozzle with the liquid waste collector；
Described device further includes nut cap, is covered on the top of sample pool, is detachably connected with the lateral wall of the liquid waste collector
Or it is detachably connected with the support edge.
6. the extracellular vesica enriching apparatus of the urine of hospital according to claim 2, which is characterized in that the filter layer
Longitudinal cross-section be V-shape；
The shape of the filter and the liquid waste collector is in cylindrical；
The outer wall of the filter and the liquid waste collector is made of transparent material；
Graduation mark is provided on the outer wall of the filter and the liquid waste collector, and these described graduation marks do not exist
On same straight line in transverse direction.
7. the extracellular vesica enriching apparatus of the urine of hospital according to claim 2, which is characterized in that described device is also
Include:
Low speed centrifuge, including low speed turntable；
Sample shifts cup, and bottom is adapted with the low speed turntable；For holding urine specimen；
The low speed centrifuge centrifugation predetermined time removes the cell fragment in the urine specimen, obtains the supernatant.
8. the extracellular vesica enriching apparatus of the urine of hospital according to claim 2, which is characterized in that the filter
It is detachably connected with the liquid waste collector；And
The bottom of the liquid waste collector and the outer wall of the liquid waste collector are detachably connected.
9. the extracellular vesica enriching apparatus of the urine of hospital according to claim 8, which is characterized in that the filter
Top and the liquid waste collector at the top of be provided with compatible shackle member；
The outer wall of the inner wall of the bottom of the liquid waste collector and the liquid waste collector is respectively arranged with corresponding inside and outside spiral shell
Line.
10. the extracellular vesica enriching apparatus of the urine of hospital according to claim 3, which is characterized in that described device
Further include:
For holding the washing cup of ultrapure water, it is set to that the liquid waste collector is external, the material of the washing cup is transparent material
Material, and graduation mark is provided on the washing cup, the capacity of the washing cup is 200 milliliters；
For blowing and beating the pipettor of the trapped fluid in the nanofiltration membrane, it is external to be set to the liquid waste collector；
For collecting the extracellular vesica collector of the extracellular vesica of the urine, it is set to the filter layer lower end, and thin
Extracellular vesica collector is removably connect with the nanometer film support frame of the filter layer；Described in piping and druming by the pipettor
The extracellular vesica of urine is retained in the extracellular vesica collector.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111007238A (en) * | 2019-11-27 | 2020-04-14 | 爱威科技股份有限公司 | Enrichment device, enrichment system, sample manufacturing system and sample detection system |
| CN112903403A (en) * | 2019-12-04 | 2021-06-04 | 中国科学院大连化学物理研究所 | Device for realizing exosome enrichment by using filter membrane and exosome enrichment method |
| CN112940910A (en) * | 2019-12-09 | 2021-06-11 | 中国科学院大连化学物理研究所 | Islet vesicle enrichment device and preparation method and application thereof |
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2018
- 2018-08-16 CN CN201821324225.2U patent/CN209485831U/en active Active
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111007238A (en) * | 2019-11-27 | 2020-04-14 | 爱威科技股份有限公司 | Enrichment device, enrichment system, sample manufacturing system and sample detection system |
| WO2021103200A1 (en) * | 2019-11-27 | 2021-06-03 | 爱威科技股份有限公司 | Enricher, enrichement system, sample manufacturing system, and sample detection system |
| CN112903403A (en) * | 2019-12-04 | 2021-06-04 | 中国科学院大连化学物理研究所 | Device for realizing exosome enrichment by using filter membrane and exosome enrichment method |
| CN112940910A (en) * | 2019-12-09 | 2021-06-11 | 中国科学院大连化学物理研究所 | Islet vesicle enrichment device and preparation method and application thereof |
| CN112940910B (en) * | 2019-12-09 | 2022-07-15 | 中国科学院大连化学物理研究所 | Islet vesicle enrichment device and preparation method and application thereof |
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