CN209485830U - The device of hospital and the home-use extracellular vesica of enrichment urine that can make - Google Patents
The device of hospital and the home-use extracellular vesica of enrichment urine that can make Download PDFInfo
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- CN209485830U CN209485830U CN201821323583.1U CN201821323583U CN209485830U CN 209485830 U CN209485830 U CN 209485830U CN 201821323583 U CN201821323583 U CN 201821323583U CN 209485830 U CN209485830 U CN 209485830U
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Abstract
Utility model device is suitable for field of medical technology, it is the device of a kind of hospital and the home-use extracellular vesica of enrichment urine that can make, the ancillary equipment other than centrifuge and other present apparatus is not needed, it can be used for the non-invasive liquid biopsy of chronic disease early diagnosis, the device includes: sample pool, it is set to the upper end of described device, the sample pool holds the urine specimen to be filtered；Nanofilter, including at least one filter layer and extracellular vesica collector；The extracellular vesica collector is set to the filter layer lower end, and the extracellular vesica collector collects the extracellular vesica of urine；Nanofilter liquid waste collector has urine impurity collecting chamber；Vacuum suction interface on the liquid waste collector side wall；Negative pressure component will be aspirated between the liquid waste collector and the nanofilter as preset negative pressure.Whereby, utility model device realizes the extracellular vesica convenient and efficient of urine and extracts.
Description
Technical field
The utility model relates to the workable enrichment urine of the field of medical instrument technology more particularly to a kind of hospital and family
The device of the extracellular vesica of liquid, the non-invasive liquid biopsy for 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 it is a kind of for the nanometer film of family and community application enrichment the extracellular vesica of urine for early diagnose it is more
The device of kind chronic 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, diameter 30-1000nm, EVs include there are many protein, lipid, various RNA (ribonucleic acid,
Ribonucleic acid) ingredient etc., it can be secreted by a variety of different types of cells, be widely distributed in various body fluid, cell can be used as
Between carrier, various physiology and pathological information are transmitted by cargo carrier function, is had and is participated in intracellular signal transduction, immune adjusts
It saves, establishes tumor escape mechanism, mediates the functions such as regeneration and degenerative process, be that there is the clinical diagnosis of high development potentiality and control
The novel targets for the treatment of, when various disease extracellular vesica secretory volume and its component content such as protein, lipid, RNA etc. can produce
Changing, these variations were both related with the pathogenesis of various disease, detected the secretory volume and component content of extracellular vesica
Difference can be used for the antidiastole of various 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.
In existing technical solution, 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 unlikely for household use in most of clinical applications, easy-to-use novel it is therefore necessary to research and develop
Technique device.
Utility model content
For above-mentioned defect, the purpose of this utility model is to provide the workable enrichment urine of a kind of hospital and family
The device of the extracellular vesica of liquid, to be implemented without the extracellular vesica of urine of ancillary equipment other than centrifuge and other present apparatus
Extraction, can be used in hospital and family.
To achieve the goals above, it is extracellular to provide the workable enrichment urine of a kind of hospital and family for the utility model
The device of vesica, comprising:
Sample pool, is set to the upper end of described device, and the sample pool holds the urine specimen to be filtered；
Nanofilter is set between the sample pool and liquid waste collector, including at least one filter layer and cell
Outer vesica collector；The extracellular vesica collector is set to the filter layer lower end, and the extracellular vesica collector is received
Collect the extracellular vesica of the urine；
Nanofilter liquid waste collector, has a urine impurity collecting chamber, and the liquid waste collector be socketed on it is described
The outside of nanofilter；Vacuum suction interface on the liquid waste collector side wall；
Negative pressure component is connected to the vacuum suction interface, will be between the liquid waste collector and the nanofilter
Suction is preset negative pressure.
According to the device, the filter layer includes:
Inceptive filtering layer, including filter membrane support frame and initial filter membrane, the initial filter membrane are embedded on filter membrane support frame；
Nanofiltration layer, including nanometer film support frame and nanofiltration membrane；The nanometer film support frame is embedded in the nanometer
The bottom of filter is connect with the side wall of the nanofilter；The extracellular vesica collector is set to the nanometer mistake
The centre of filtering layer, and the extracellular vesica collector is detachably connected with the nanometer film support frame.
According to the device, the vacuum suction interface is set to the side wall upper part of the liquid waste collector, described true
Suction gas interface is for connecting negative pressure component；
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 device, the filter membrane support frame includes multiple first support items, one end of multiple first support items
It is connect respectively with the bottom with the side wall of the sample pool, the other end links together；
The nanometer film support frame includes multiple second support items, one end of multiple second support items respectively with described and institute
The bottom connection of the side wall of nanofilter is stated, the other end links together.
According to the device, the initial filter membrane have multiple first filter hole, first filter hole aperture it is big
Small is 2 microns；The nanofiltration membrane has multiple second filter hole, and the size in multiple second filter hole apertures is received for 20
Rice；
The capacity of the extracellular vesica collector is 5~20 milliliters.
According to the device, the inceptive filtering layer is that filtering includes cell fragment and the urine greater than extracellular vesica
The inceptive filtering layer of liquid impurity；The nanofiltration membrane layer is the urine cell external capsule that retention is greater than the specific filtration aperture of nanometer film
Bubble, and filter the nanofiltration membrane layer of the urine impurity including soluble protein etc.；By the inceptive filtering layer and the nanometer
The extracellular vesica of the urine that filter membranous layer filtering obtains is collected in the extracellular vesica collector.
According to the device, the longitudinal cross-section of the inceptive filtering layer is V-shape or circular arc type；The nanofiltration
The longitudinal cross-section of layer is V-shape or circular arc type.
According to the device, the shape of the sample pool, nanofilter and the liquid waste collector is in cylinder
The bodily form；
The outer wall of the sample pool, nanofilter and the liquid waste collector is made of transparent material；
The bottom of the liquid waste collector and the outer wall of the liquid waste collector are detachably connected.
According to the device, it is respectively provided on the outer wall of the sample pool, nanofilter and the liquid waste collector
There is graduation mark, and on the same straight line of these described graduation marks not in a lateral direction；
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 device, 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；And
The negative pressure component is vacuum pump.
The utility model is by the way that the device of the workable enrichment extracellular vesica of urine of hospital and family to be set as wrapping
Sample pool, nanofilter and liquid waste collector are included, the bottom of nanofilter is equipped with nanometer film, is formed inside nanofilter
The storage chamber of EVs rich nanometer forms the waste collection chamber of urine impurity between liquid waste collector inside and nanofilter lower section,
The tip edge of nanofilter be equipped with support edge, the support along between the nozzle of liquid waste collector top be enclosed construction, waste collection
Device side wall is equipped with vacuum pump air-breathing interface tube, is controlled to a vacuum pump by air intake duct.Thus, it is possible to carry out negative pressure by vacuum pump
Pumping, is filtered urine specimen using hydrostatic pressing and filter membrane pressure at both sides, obtains EVs, and is not necessarily to complex operations, without from
Other laboratory apparatus and reagent processing other than scheming and the present apparatus, it is simple and quick.The present apparatus is easy to carry, using consumptive material
Cost price is cheap, is suitble to use in family and community hospital's large-scale promotion.
Detailed description of the invention
Fig. 1 is the dress of the workable enrichment extracellular vesica of urine of hospital and family provided by the embodiment of the utility model
Set structural schematic diagram；
Fig. 2 is the dress of the workable enrichment extracellular vesica of urine of hospital and family provided by the embodiment of the utility model
Set structural schematic diagram；
Fig. 3 is the apparatus structure schematic diagram of the workable enrichment extracellular vesica of urine of hospital and family shown in Fig. 2；
Fig. 4 A, which is that hospital and family provided by the embodiment of the utility model are workable, is enriched with the extracellular vesica of urine
Filter membrane support frame structure diagram in device；
Fig. 4 B, which is that hospital and family provided by the embodiment of the utility model are workable, is enriched with the extracellular vesica of urine
Filter membrane support frame structure diagram in device；
Fig. 5 A, which is that hospital and family provided by the embodiment of the utility model are workable, is enriched with the extracellular vesica of urine
Nanometer film support frame structure diagram in device；
Fig. 5 B, which is that hospital and family provided by the embodiment of the utility model are workable, is enriched with the extracellular vesica of urine
Nanometer film support frame structure diagram in device；
Fig. 6 is the dress of the workable enrichment extracellular vesica of urine of hospital and family provided by the embodiment of the utility model
Set the inspection result schematic diagram of the enrichment extracellular vesica of urine.
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 hospital is provided in one embodiment of the utility model and family is workable
It is enriched with the device 100 of the extracellular vesica of urine, comprising:
Sample pool 10, is set to the upper end of described device, and the sample pool 10 holds the urine specimen to be filtered；
Nanofilter 20 is set between the sample pool 10 and liquid waste collector 30, including at least one filter layer
With extracellular vesica collector 23；The filter layer is connect with the extracellular vesica collector 23, extracellular vesica collector
23 are set to filter layer lower end, and extracellular vesica collector 23 collects the extracellular vesica of the urine；
Liquid waste collector 30 has urine impurity collecting chamber, and the liquid waste collector 30 is socketed on the nanometer mistake
The outside of filter 20；Vacuum suction interface 315 on 30 side wall of liquid waste collector；
Negative pressure component is connected to the vacuum suction interface 315, will take out between liquid waste collector 30 and nanofilter 20
Inhaling is preset negative pressure.
In this embodiment, the device 100 of the workable enrichment extracellular vesica of urine of hospital and family includes sample
Pond 10, nanofilter 20 and liquid waste collector 30.Wherein sample pool 10 is for placing the urine specimen to be filtered；It receives
Rice filter 20 includes at least one filter layer and extracellular vesica collector 23, and filter layer has filter membrane, can filter out
The extracellular vesica of urine is extracted, and is stored in extracellular vesica collector 23 by the urine impurity in urine specimen.The hospital
And the device 100 of the workable enrichment extracellular vesica of urine of family is low in cost, easy to operate, time-consuming short, gained cell
Outer vesica yield and purity is high and structural integrity, without the other instruments equipment other than centrifuge and the present apparatus, for family and
Community's large-scale promotion uses.The device 100 of the workable enrichment extracellular vesica of urine of hospital and family for family and
Community application, by the further detection and analysis that can be used for early diagnosing a variety of chronic diseases to the extraction of extracellular vesica.Change pre-
If negative pressure be subject to be able to achieve enrichment purpose.
Referring to FIG. 1 to FIG. 3, in one embodiment of the utility model, filter layer 21 includes:
Inceptive filtering layer 21, including filter membrane support frame 211 and initial filter membrane 222；Filter membrane support frame 211 and sample pool 10
Bottom connection, initial filter membrane 222 are embedded on filter membrane support frame 211；
Nanofiltration layer 22, including nanometer film support frame 212 and nanofiltration membrane 213；Nanometer film support frame 212, which is set to, to be received
The bottom of rice filter 20, connect with the side wall of nanofilter 20；Nanofiltration membrane 213 is embedded on nanometer film support frame 212；
Extracellular vesica collector 23 is set to the centre of nanofiltration layer 22；And extracellular vesica collector 23 and nanometer film branch
Support 212 is detachably connected, and the top of extracellular vesica collector 23 is such as provided with resilient clamp member, and is supported in nanometer film
The centre of frame 212 is provided with clamping corresponding with resilient clamp member position, it is possible thereby to easily receive extracellular vesica
The installation of storage 23 and taking-up.The bottom of sample cell 10 is to the inclined filter membrane support frame 211 in middle part, 211 inside of filter membrane support frame
Embedded with initial filter membrane 222；The bottom of nanofilter 20 is to the inclined nanometer film support frame 212 in middle part, nanometer film support frame
212 insides are embedded with nanofiltration membrane 213, and the central Formation cross-section of nanofilter 20 is the extracellular vesica of the EVs of the cone of V-arrangement
Collector 23；30 outer wall of liquid waste collector of nanofilter 20 and vacuum chamber is equipped with volume scale.
In addition, the side wall upper part of liquid waste collector 30 is provided with vacuum suction interface 315, vacuum suction interface 315 is used for
Connect vacuum pump；The top of device 100 has support along 32, and holding in the palm the top nozzle along 32 and liquid waste collector 30 is closing knot
Structure.There is an enclosed construction at initial filter membrane 222 and the edge of nanofiltration membrane 213, and 20 top nozzle of nanofilter is equipped with support along 32,
The support is enclosed construction between 32 and 30 top nozzle of liquid waste collector, and the 30 top nozzle of liquid waste collector of vacuum chamber is equipped with lid
Cap, tube wall upper semisection outside are equipped with the vacuum suction interface 315 of negative pressure component connection, and negative pressure component can be vacuum pump, lead to
It crosses vacuum suction interface 315 to dock with vacuum pump, can will will be aspirated between liquid waste collector 30 and nanofilter 20
For preset negative pressure, increase filter efficiency.In addition, the device of the workable enrichment extracellular vesica of urine of hospital and family
100 further include nut cap 50, is covered on the top of sample pool 10, be detachably connected with the lateral wall of the liquid waste collector 30 or
It is detachably connected with the support along 32.For example, can be spirally connected with the lateral wall of liquid waste collector 30, or with support along 32 cards
It connects, is provided with support in the inside of nut cap 50 along 32 compatible seal gaskets.It is possible thereby to which hospital and family can be used
The extracellular vesica of enrichment urine device 100 it is closed, form the environment vacuumized.When needing using the hospital and family
When the device 100 of the extracellular vesica of enrichment urine used is enriched with urine extracellular vesica, after sample liquid is added, by nut cap 50 with
The lateral wall of liquid waste collector 30 screws, and after the completion of enrichment, then unscrews nut cap 50, carries out the cleaning etc. of device 100.Figure
3 be the structure letter not including nut cap of the device of the workable enrichment extracellular vesica of urine of hospital and family shown in Fig. 2
Figure.In addition, referring to fig. 2 and Fig. 3, the bottom 301 of liquid waste collector and the outer wall of liquid waste collector 30 are detachably connected.The two can
It is attached by by way of the shackle member of adaptation either threaded connection.Specifically the bottom 301 of liquid waste collector is interior
The outer wall of wall and liquid waste collector 30 is respectively arranged with corresponding internal and external screw thread.It, can be by that will give up after the completion of enrichment process
The bottom 301 of collection is dismantled, and is then dismantled extracellular vesica collector 23, is taken out the extracellular vesica of the urine being enriched to.
A and Fig. 4 B referring to fig. 4, in one embodiment of the utility model, the filter membrane support frame 211 includes multiple the
One bracket item 2111, the connection with the side wall of the bottom of the sample pool 10 respectively of one end of multiple first support items 2111,
The other end links together；Inceptive filtering layer 21 has certain gradient as a result, can be very good to be filtered；Multiple
One bracket item 2111 is connect with the side wall of the bottom of sample pool 10, will constitute a pyramid type, such as Fig. 4 A or a circular arc
Type, such as Fig. 4 B.
Referring to Fig. 5 A and Fig. 5 B, the nanometer film support frame 212 includes multiple second support items 2121, multiple second supports
One end of item 2121 is connect with the bottom of the side wall of nanofilter 20 respectively, and the other end links together.Multiple second
Frame item 2121 is connect with the bottom of the side wall of nanofilter 20, will constitute a pyramid type or a circular arc type, is such as schemed
5A or circular arc type, such as Fig. 5 B.
Specifically, initial filter membrane 222 has multiple first filter hole, the diameter of first filter hole is 2 microns；
Nanofiltration membrane 213 has multiple second filter hole, and the diameter of multiple second filter hole is 20 nanometers；Extracellular vesica
The capacity of collector 23 is 5~20 milliliters.Preferably, it includes that cell fragment and volume are greater than cell that inceptive filtering layer 21, which is filtering,
The inceptive filtering layer of other urine impurity of outer vesica；The nanofiltration membrane layer 213 is that can filter including soluble component, small point
Sub- albumen equal diameter is less than the nanofiltration membrane layer of the urine impurity of extracellular vesica.；By inceptive filtering layer 21 and nanofiltration membrane
The extracellular vesica of the urine that 22 filtering of layer obtains is collected in the extracellular vesica collector 23.Extracellular vesica is collected
The connection of device 23 and 212 bottom of nanometer film support frame can be and connect with multiple 2121 common connection end of second support item, and
And the connecting pin is annular in shape, convenient for the extracellular vesica of urine drippage in extracellular vesica collector 23.
In one embodiment of the utility model, sample is first poured into sample cell 10, outside vacuum chamber liquid waste collector 30
The vacuum pump air-breathing interface tube of side connects upper vacuum pump and carries out negative pressure ventilation, sample poor using hydrostatic pressing and filter membrane pressure at both sides
Cell fragment can be removed by initial filter membrane 222 and volume is greater than other urine impurity of extracellular vesica；Then sample reaches
In nanofilter 20, by EVs of the diameter at 30 to 1000 nanometers can be retained when nanofiltration membrane 213；The EVs being finally enriched with will
It is collected into the EVs collector (extracellular vesica collector 23) of V-arrangement, that is, completes the enrichment of the extracellular vesica of urine.This reality
With novel low in cost, easy to operate, time-consuming short, the extracellular vesica yield of gained and with high purity and structural integrity, without centrifugation
Other instruments equipment other than machine and the present apparatus, can large-scale promotion the advantages that using for family and community.
In one embodiment of the utility model, the longitudinal cross-section of inceptive filtering layer 21 is V-shape or circular arc type；Institute
The longitudinal cross-section for stating nanofiltration layer 22 is V-shape or circular arc type.Sample pool 10, nanofilter 20 and the waste liquid are received
The shape of storage 30 is in cylindrical；The outer wall of sample pool 10, nanofilter 20 and the liquid waste collector 30 is by saturating
Bright material is made.Graduation mark, and this are provided on the outer wall of sample pool 10, nanofilter 20 and liquid waste collector 30
On a little same straight lines of the graduation mark not in a lateral direction.Thus, it is possible to be clearly seen that sample pool 10, nanofilter
20 and liquid waste collector 30 outer wall on scale, and the capacity of solution loaded in current each container.
Furthermore the device 100 of the workable enrichment extracellular vesica of urine of hospital and family further include:
For holding the washing cup of ultrapure water, it is set to the liquid waste collector 30 in vitro, the material of the washing cup is
Transparent 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 213, it is external to be set to the liquid waste collector 30.
Device 100 using the workable enrichment extracellular vesica of urine of hospital provided by the utility model and family is rich
Collecting the extracellular vesica of urine, the specific method is as follows:
It is added portionwise in sample cell after sample collection according to 10 volume of sample cell, 30 outside of liquid waste collector of vacuum chamber
Vacuum suction interface 315 connects upper vacuum pump and carries out negative pressure ventilation, and poor using hydrostatic pressing and filter membrane pressure at both sides, sample is through via hole
The initial filter membrane 222 that diameter is 2 microns can remove cell fragment and volume is greater than other urine impurity of extracellular vesica.Sample arrives
Up in nanofilter 20, diameter can be retained at 30 to 1000 nanometers when by aperture being 20 nanometers of nanofiltration membrane 213
EVs, and remaining soluble protein etc. is then oozed out from nanofiltration membrane 213, into vacuum chamber liquid waste collector 30.To nanometer mistake
When trapped fluid residue 10ml in filter 20 or so, 200ml ultrapure water is added into EVs Nano-enricher with washing cup, continues
Above-mentioned nanometer film process.When the trapped fluid residue 5ml in nanofilter 20, mix it simultaneously with pipettor piping and druming trapped fluid
It is collected into the extracellular vesica collector 23 of EVs, which is the extracellular vesicle solution of final urine.Then it can incite somebody to action
The bottom of liquid waste collector 30 is dismantled, and is finally taken out extracellular vesica collector 23, will be obtained last urine cell external capsule
Steep detection, such as nephrosis, diabetes etc. that solution is used for chronic disease.
(1) the initial membrane filtration that pore size is 2 microns: taking the Urine specimens of 30ml healthy volunteer, studied, will
Urine specimens first pour into sample cell, and the vacuum pump air-breathing interface tube on the outside of vacuum chamber liquid waste collector connects upper vacuum pump and born
Pressure pumping, poor using hydrostatic pressing and filter membrane pressure at both sides, sample passes through initial filter membrane, removes cell fragment and volume is greater than cell
Other urine impurity of outer vesica；
(2) nanofiltration membrane that pore size is 20 nanometers filters: sample reaches in nanofilter, can by nanofiltration membrane
Retain EVs of the diameter at 30 to 1000 nanometers；
(3) enrichment of EVs: when the trapped fluid residue 10ml or so in nanofilter, with washing cup toward EVs rich nanometer
200ml ultrapure water is added in storage, continues above-mentioned nanometer film process.When the trapped fluid residue 5ml in nanofilter, with shifting
Liquid device piping and druming trapped fluid mixes it and is collected into the extracellular vesica collector of EVs, which is that final urine is thin
Extracellular vesicle solution.The EVs being finally enriched with will be collected into the EVs collector (extracellular vesica collector) of V-arrangement, that is, be completed
The enrichment of the extracellular vesica of urine.
Initial filter membrane and nanofiltration membrane can be used must pure (Shanghai) trade Co., Ltd Spectrum Labs purchased from bodyguard
Inc, the initial filter membrane that pore size is 2 microns removes cell fragment and volume is greater than other urine impurity of extracellular vesica,
Nanofiltration membrane molecular cut off is 1000KDa, retains EV 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 the extracellular vesica of urine, as shown in concrete outcome Fig. 6, from fig. 6, it can be seen that urine cell external capsule
Steep cup-shaped or round, partial size is in 30-1000nm.
(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. 6.
The mean particle size of the extracellular vesica of urine is 102nm.
It is provided by the utility model compared with existing common supercentrifugation, magnetic bead absorption method and isolation kit method
Extracellular having the advantage that for vesica can be enriched with acquisition and the comparable extracellular vesica of above method purity, be not necessarily to centrifuge
With the other instruments equipment other than the present apparatus, can be retained when because aperture being 20 nanometers of nanofiltration membrane 213 in the present apparatus 100 straight
EVs of the diameter at 30 to 1000 nanometers, and remaining soluble protein etc. is then oozed out from nanofiltration membrane 213.The present apparatus 100 can be primary
Property a large amount of urine specimens of processing, and be not necessarily to complex operations.Laboratory or even family of the present apparatus 100 configured with minipump
I.e. can be used, without other than centrifuge and the present apparatus other laboratory apparatus and reagent processing.The present apparatus 100 by vacuum pump into
Row negative pressure ventilation is realized using hydrostatic pressing and filter membrane pressure at both sides difference, simple and quick.The present apparatus 100 is easy to carry, adopts
It is cheap with consumables cost, it is suitble to use in family and community hospital's large-scale promotion.Wherein initial filter membrane 222 and nanometer filter
The miillpore filter production in corresponding aperture is respectively adopted in film 213, as nanofiltration membrane 213 uses aperture for 20 nanometers of nanometer film system
Make, the molecular cut off of the nanometer film is 1000kDa.
The utility model is by the way that the device of the workable enrichment extracellular vesica of urine of hospital and family to be set as wrapping
Sample pool, nanofilter and liquid waste collector are included, the bottom of nanofilter is equipped with nanometer film, is formed inside nanofilter
The storage chamber of EVs rich nanometer forms the waste collection chamber of urine impurity between liquid waste collector inside and nanofilter lower section,
The top nozzle of nanofilter be equipped with support edge, the support along between the nozzle of liquid waste collector top be enclosed construction, tube wall upper half
Section outside is equipped with vacuum pump air-breathing interface tube, is controlled to a vacuum pump by air intake duct.Thus, it is possible to carry out negative pressure by vacuum pump
Pumping, is filtered urine specimen using hydrostatic pressing and filter membrane pressure at both sides, obtains EVs, and is not necessarily to complex operations, without from
Other laboratory apparatus and reagent processing other than scheming and the present apparatus, it is simple and quick.The present apparatus is easy to carry, using consumptive material
Cost price is cheap, is suitble to use in family and community hospital's large-scale promotion.
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. the device of a kind of hospital and the home-use extracellular vesica of enrichment urine that can make characterized by comprising
Sample pool, is set to the upper end of described device, and the sample pool holds the urine specimen to be filtered；
Nanofilter is set between the sample pool and liquid waste collector, including at least one filter layer and cell external capsule
Steep collector；The extracellular vesica collector is set to the filter layer lower end, and the extracellular vesica collector collects institute
State the extracellular vesica of urine；
Nanofilter liquid waste collector has urine impurity collecting chamber, and the liquid waste collector is socketed on the nanometer
The outside of filter；Vacuum suction interface on the liquid waste collector side wall；
Negative pressure component is connected to the vacuum suction interface, will aspirate between the liquid waste collector and the nanofilter
For preset negative pressure.
2. the device of hospital according to claim 1 and the home-use extracellular vesica of enrichment urine that can make, feature
It is, the filter layer includes:
Inceptive filtering layer, including filter membrane support frame and initial filter membrane, the initial filter membrane are embedded on filter membrane support frame；
Nanofiltration layer, including nanometer film support frame and nanofiltration membrane；The nanometer film support frame is embedded in the nanofiltration
The bottom of device is connect with the side wall of the nanofilter；The extracellular vesica collector is set to the nanofiltration layer
Centre, and the extracellular vesica collector is detachably connected with the nanometer film support frame.
3. the device of hospital according to claim 2 and the home-use extracellular vesica of enrichment urine that can make, feature
It is, the vacuum suction interface is set to the side wall upper part of the liquid waste collector, and the vacuum suction interface is for connecting
Negative pressure component；
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 the sample pool, detachable with the lateral wall of the liquid waste collector
Connection is detachably connected with the support edge.
4. the device of hospital according to claim 2 and the home-use extracellular vesica of enrichment urine that can make, feature
Be, the filter membrane support frame includes multiple first support items, one end of multiple first support items respectively with the sample pool
The bottom of side wall connects, and the other end links together；
The nanometer film support frame includes multiple second support items, one end of multiple second support items respectively with the nanofiltration
The bottom of the side wall of device connects, and the other end links together.
5. the device of hospital according to claim 2 and the home-use extracellular vesica of enrichment urine that can make, feature
It is, the initial filter membrane has multiple first filter hole, and the size in first filter hole aperture is 2 microns；The nanometer
Filter membrane has multiple second filter hole, and the size in multiple second filter hole apertures is 20 nanometers；
The capacity of the extracellular vesica collector is 5~20 milliliters.
6. the device of hospital according to claim 5 and the home-use extracellular vesica of enrichment urine that can make, feature
It is, the inceptive filtering layer is the inceptive filtering layer that filtering includes cell fragment and the urine impurity greater than extracellular vesica；
The nanofiltration layer is the extracellular vesica of urine that retention is greater than nanometer film filtration aperture, and filtering includes soluble protein
The nanofiltration layer of urine impurity；The urine cell obtained by the inceptive filtering layer and nanofiltration layer filtering
Outer vesica is collected in the extracellular vesica collector.
7. the device of hospital according to claim 2 and the home-use extracellular vesica of enrichment urine that can make, feature
It is, the longitudinal cross-section of the inceptive filtering layer is V-shape or circular arc type；The longitudinal cross-section of the nanofiltration layer is V word
Type or circular arc type.
8. the device of hospital according to claim 1 and the home-use extracellular vesica of enrichment urine that can make, feature
It is, the shape of the sample pool, nanofilter and the liquid waste collector is in cylindrical；
The outer wall of the sample pool, nanofilter and the liquid waste collector is made of transparent material；
The bottom of the liquid waste collector and the outer wall of the liquid waste collector are detachably connected.
9. the device of hospital according to claim 8 and the home-use extracellular vesica of enrichment urine that can make, feature
It is, graduation mark, and these institutes is provided on the outer wall of the sample pool, nanofilter and the liquid waste collector
It states on the same straight line of graduation mark not in a lateral direction；
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 device of hospital according to claim 3 and the home-use extracellular vesica of enrichment urine that can make, feature
It is, 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；And
The negative pressure component is vacuum pump.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821323583.1U CN209485830U (en) | 2018-08-16 | 2018-08-16 | The device of hospital and the home-use extracellular vesica of enrichment urine that can make |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201821323583.1U CN209485830U (en) | 2018-08-16 | 2018-08-16 | The device of hospital and the home-use extracellular vesica of enrichment urine that can make |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112834305A (en) * | 2021-02-01 | 2021-05-25 | 深圳褀氏生物医疗电子有限公司 | Method and device for efficiently separating enriched cells through continuous flow |
| 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 |
-
2018
- 2018-08-16 CN CN201821323583.1U patent/CN209485830U/en active Active
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN112834305A (en) * | 2021-02-01 | 2021-05-25 | 深圳褀氏生物医疗电子有限公司 | Method and device for efficiently separating enriched cells through continuous flow |
| CN112834305B (en) * | 2021-02-01 | 2022-01-11 | 深圳褀氏生物医疗电子有限公司 | Method and device for efficiently separating enriched cells through continuous flow |
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