CN107702973A - A kind of whole blood blood plasma piece-rate system and method - Google Patents
A kind of whole blood blood plasma piece-rate system and method Download PDFInfo
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- CN107702973A CN107702973A CN201710804368.7A CN201710804368A CN107702973A CN 107702973 A CN107702973 A CN 107702973A CN 201710804368 A CN201710804368 A CN 201710804368A CN 107702973 A CN107702973 A CN 107702973A
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Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5025—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures for parallel transport of multiple samples
- B01L3/50255—Multi-well filtration
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
Abstract
The present invention provides a kind of whole blood blood plasma piece-rate system and method, wherein, system includes the first blood separating mechanism and the second blood separating mechanism, first blood separating mechanism includes blood collection unit and at least one first micro-fluidic chip, micro-pillar array is provided with first micro-fluidic chip, the minimum intercolumniation of micro-pillar array is more than or equal to the diameter of red blood cell, second blood separating mechanism includes the second micro-fluidic chip, blood plasma recovery unit and devil liquor recovery unit, and fluid channel is provided with the second micro-fluidic chip；Blood collection unit, at least one first micro-fluidic chip and two micro-fluidic chips are arranged in order along blood flow direction and connected from beginning to end successively by fluid line, blood plasma recovery unit is connected by fluid line and the plasma outlet port of fluid channel, and devil liquor recovery unit is connected by fluid line and the waste liquid outlet of fluid channel.The present invention can realize the detection efficiency for haemocyte in minimal amount of blood sample and hematoblastic quick separating, improving blood sample.
Description
Technical field
The embodiment of the present invention belongs to field of biomedicine technology, more particularly to a kind of whole blood blood plasma piece-rate system and method.
Background technology
Blood is made up of haemocyte and blood plasma, and the histoorgan of blood and each system of whole body contacts with close,
Under normal physiological conditions, the level of the quality and quantity of various composition in blood, the eubolism and body of body are directly reflected
The poised state of internal and external environment；In pathological conditions, blood is in addition to it can directly reflect the illness of human body hemopoietic system, moreover it is possible to
Directly or indirectly react the lesion of whole body or local organization organ.Therefore, blood not only serves as primary hemopoietic system disease
The diagnosis of disease, antidiastole, the Main Basiss of observation of curative effect and Index for diagnosis, and can also be to cause Secondary cases blood change
The diagnosis and treatment of other each systemic diseases provide important information.Under normal circumstances, carrying out blood testing needs the haemocyte in blood
Separate, because spectrum analysis of its presence to blood testing has very big interference.
Generally all it is using large-scale however, at present during clinical diagnosis and basic research of the overwhelming majority to blood
Centrifuge come separated plasma and haemocyte, it is necessary to expend the substantial amounts of blood of patient, and centrifuge volume is larger, structure is multiple
It is miscellaneous, it is impossible to be directly connected to blood testing equipment so that blood separates and blood testing needs to be separated and independently performed, and easily causes sample
Product pollute, and detection efficiency is low, it is impossible to meet safety, quick blood testing demand.
The content of the invention
The present invention can carry out quick separating to minimal amount of blood sample, can be directly connected to blood testing equipment, improve
The detection efficiency of blood sample, it is small volume, simple in construction, complete, quick blood testing requirement can be met.
On the one hand the embodiment of the present invention provides a kind of whole blood blood plasma piece-rate system, it includes the first blood separating mechanism and the
Two blood separating mechanisms, first blood separating mechanism include blood collection unit and at least one first micro-fluidic chip,
Micro-pillar array is provided with first micro-fluidic chip, the minimum intercolumniation of the micro-pillar array is more than or equal to red blood cell
Diameter, second blood separating mechanism include the second micro-fluidic chip, blood plasma recovery unit and devil liquor recovery unit, and described
Fluid channel is provided with two micro-fluidic chips；
The blood collection unit, at least one first micro-fluidic chip and two micro-fluidic chip are along blood stream
To being arranged in order and being connected from beginning to end successively by fluid line, the blood plasma recovery unit passes through fluid line and the fluid channel
Plasma outlet port connection, the devil liquor recovery unit is connected by fluid line with the waste liquid outlet of the fluid channel；
The blood of blood collection unit collection flows at least one first micro-fluidic chip successively, it is described at least
One the first micro-fluidic chip retains to the haemocyte in the blood, obtains containing blood platelet and red less than predetermined amount
The blood plasma of cell；It is small that second micro-fluidic chip obtains the blood plasma of high-purity, blood to blood plasma progress inertia focusing
Plate and the red blood cell less than predetermined amount, the blood plasma of the high-purity flow into the blood plasma recovery unit, the blood platelet and are less than
The red blood cell of predetermined amount is through flowing into the devil liquor recovery unit.
In one embodiment, the micro-pillar array includes at least one sub- microtrabeculae battle array being arranged in order along blood flow direction
Row, the intercolumniation of each sub- micro-pillar array differ, and the intercolumniation of each sub- micro-pillar array is according at least one son
Putting in order for micro-pillar array is sequentially reduced, and the micro-pillar array, which comprises at least, is arranged in the sub- microtrabeculae battle array of least significant end red blood cell retention
Row, the red blood cell retain diameter of the intercolumniation more than or equal to red blood cell of sub- micro-pillar array；
The red blood cell retains sub- micro-pillar array and the haemocyte in the blood is retained, to filter in the blood
Haemocyte.
In one embodiment, the micro-pillar array is in cylindrical-array, cylindroid array or polygon pillar array
Any, the diameter range of the microtrabeculae in the micro-pillar array is 10 microns~200 microns, and the altitude range 10 of the microtrabeculae is micro-
Rice~200 microns.
In one embodiment, the whole blood blood plasma piece-rate system also includes minitype pneumatic valve, the minitype pneumatic valve
Air inlet connects constant pressure source of the gas, and the gas outlet of the minitype pneumatic valve connects the entrance of the blood collection unit；
When the minitype pneumatic valve is opened, output constant pressure source of the gas is promoted to the blood collection unit by gas pressure
Blood in the blood collection unit flows into first micro-fluidic chip.
In one embodiment, the whole blood blood plasma piece-rate system also includes blood plasma buffer pool, the blood plasma buffer pool
Entrance is connected by fluid line with the outlet for first micro-fluidic chip for being arranged in least significant end, the blood buffer pond
Outlet is connected by fluid line with the entrance of second micro-fluidic chip；
The blood plasma for being arranged in first micro-fluidic chip output of least significant end flows into the blood plasma buffer pool, the blood plasma
Buffer pool stores to the blood plasma, when the blood plasma exceedes the memory capacity of the blood plasma buffer pool, flows into described second
Micro-fluidic chip.
In one embodiment, the fluid channel includes multiple micro-channel units of periodic arrangement, the multiple miniflow
Road unit connects from beginning to end successively, and the micro-channel unit includes the first semi-circular fluid channel and the second semi-circular fluid channel, described
The outlet of first semi-circular fluid channel and the entrance slitless connection of the second semi-circular fluid channel.
In one embodiment, the difference of the external diameter of the first semi-circular fluid channel and internal diameter is equal to first semi-circular
The ring cutting diameter at any place in fluid channel, the external diameter of the second semi-circular fluid channel and the difference of internal diameter are less than second semi-ring
The maximum ring cutting diameter of shape fluid channel, the external diameter of the first semi-circular fluid channel are less than the interior of the second semi-circular fluid channel
Footpath.
In one embodiment, the quantitative range of the micro-channel unit is 1~50, the first semi-circular fluid channel
Ring cutting diameter range with the second semi-circular fluid channel is 1 micron~200 microns.
In one embodiment, the plasma outlet port is arranged on above the waste liquid outlet.
On the other hand the embodiment of the present invention provides a kind of whole blood plasma separation method, including：
Blood is gathered, the blood includes whole blood blood plasma；
The micro-pillar array for being more than or equal to the diameter of red blood cell by minimum intercolumniation is entered to the haemocyte in the blood
Row retains at least once, obtains the blood plasma of the red blood cell containing blood platelet and less than predetermined amount；
Inertia focusing is carried out to the blood plasma by the micro-fluidic chip for being provided with fluid channel, further described in filtering
Blood platelet and the red blood cell less than predetermined amount, obtain the blood plasma of high-purity；
Individually reclaim the blood plasma of the high-purity；
Reclaim the blood platelet and the red blood cell less than predetermined amount simultaneously.
System provided in an embodiment of the present invention passes through the micro-pillar array including intercolumniation more than or equal to the diameter of red blood cell
The first blood separating mechanism initial gross separation is carried out to the haemocyte in blood, pass through the second blood including fluid channel separation dress
Put and inertia focusing is carried out to blood, the further blood platelet and residual red blood cells separated in blood, it is possible to achieve to pole
Haemocyte and hematoblastic quick separating in a small amount of blood sample, blood testing equipment can be directly connected to, improve blood sample
Detection efficiency, it is small volume, simple in construction, complete, quick blood testing requirement can be met.
Brief description of the drawings
Technical scheme in order to illustrate the embodiments of the present invention more clearly, make required in being described below to embodiment
Accompanying drawing is briefly described, it should be apparent that, drawings in the following description are some embodiments of the present invention, for ability
For the those of ordinary skill of domain, on the premise of not paying creative work, it can also be obtained according to these accompanying drawings other attached
Figure.
Fig. 1 is the structural representation for the whole blood blood plasma piece-rate system that one embodiment of the present of invention provides；
Fig. 2 is the top view for the first micro-fluidic chip that one embodiment of the present of invention provides；
Fig. 3 is the front view for the second micro-fluidic chip that one embodiment of the present of invention provides；
Fig. 4 is the front view for the fluid channel that one embodiment of the present of invention provides；
Fig. 5 is the structural representation for the whole blood blood plasma piece-rate system that an alternative embodiment of the invention provides；
Fig. 6 is the structural representation for the whole blood blood plasma piece-rate system that yet another embodiment of the present invention provides；
Fig. 7 is the schematic flow sheet for the whole blood plasma separation method that one embodiment of the present of invention provides.
Embodiment
In order that those skilled in the art more fully understand the present invention program, below in conjunction with the embodiment of the present invention
Accompanying drawing, the technical scheme in the embodiment of the present invention is explicitly described, it is clear that described embodiment is the present invention one
The embodiment divided, rather than whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are not doing
Go out under the premise of creative work the every other embodiment obtained, should all belong to the scope of protection of the invention.
Term " comprising " and their any deformations in description and claims of this specification and above-mentioned accompanying drawing, meaning
Figure is to cover non-exclusive include.Such as process, method or system comprising series of steps or unit, product or equipment do not have
The step of being defined in the step of having listed or unit, but alternatively also including not listing or unit, or alternatively also wrap
Include for the intrinsic other steps of these processes, method, product or equipment or unit.In addition, term " first ", " second " and
" 3rd " etc. is to be used to distinguish different objects, not for description particular order.
As shown in figure 1, one embodiment of the present of invention provides a kind of whole blood blood plasma piece-rate system 100, it includes the first blood
The blood separating mechanism 20 of liquid separating apparatus 10 and second.
In the present embodiment, the first blood separating mechanism is mainly used in pre-processing blood, separates and retain blood
In haemocyte, the haemocyte in blood mainly includes red blood cell, in addition to monocyte and leucocyte, comes for tumor patient
Say, tumour cell is also possible that in blood.The diameter ＞ of the diameter ＞ leucocytes of the diameter ＞ monocytes of tumour cell is red
The diameter of cell.Haemocyte in blood there remains comprising hematoblastic blood plasma after being separated, the second blood separating mechanism
Effect is the blood platelet in further separation blood, and the blood plasma for obtaining high-purity is used to export to blood testing equipment progress
Blood testing.
As shown in figure 1, in the present embodiment, the first blood separating mechanism 10 includes blood collection unit 11 and at least one
First micro-fluidic chip 12, micro-pillar array 13 is provided with the first micro-fluidic chip 12, the minimum intercolumniation of micro-pillar array 13 is big
In or equal to red blood cell diameter.
In a particular application, the quantity of the first micro-fluidic chip can be set according to being actually needed, the first micro-fluidic chip
The more separating and filtering effects to haemocyte of quantity it is better, but quantity excessively also result in haemocyte separating rate drop
It is low, therefore, should be according to the separating and filtering effect requirements and blood to haemocyte so as to reduce overall blood testing efficiency
Detection efficiency requirement carrys out comprehensive analysis, selects an appropriate number of first micro-fluidic chip, while separating and filtering effect is ensured,
Also separative efficiency is ensured.It is exemplary in Fig. 1 that two the first micro-fluidic chips are shown.
In one embodiment, the quantitative range of the first micro-fluidic chip is 1~10.
In a particular application, because the minimum cell of diameter in haemocyte is red blood cell, therefore it may only be necessary to make micro-pillar array
Minimum intercolumniation be more than or equal to the diameter of red blood cell, it becomes possible to realize filtering to all haemocytes in blood.
In the present embodiment, intercolumniation is specifically referred in micro-pillar array, is being flowed to blood on vertical direction, any phase
The width in the space between two adjacent microtrabeculaes, the intercolumniation in the present embodiment specifically refer to horizontal intercolumniation.
In a particular application, in micro-pillar array, flowed to blood on parallel direction, two microtrabeculaes of arbitrary neighborhood it
Between the width (i.e. longitudinal intercolumniation) in space can be according to setting be actually needed, longitudinal intercolumniation can be equal to horizontal intercolumniation
Away from.
In one embodiment, the diameter range of the microtrabeculae in micro-pillar array is 10 microns~200 microns, the height of microtrabeculae
10 microns~200 microns of scope.
In one embodiment, a diameter of 30 microns of microtrabeculae, the height of microtrabeculae is 40 microns.
In a particular application, micro-pillar array can be appointing in cylindrical-array, cylindroid array or polygon pillar array
It is a kind of.The exemplary top view for showing micro-pillar array in Fig. 1, the micro-pillar array shown in Fig. 1 are cylindrical-array.
In the present embodiment, blood is obtained containing blood platelet and less than predetermined amount after the separation of the first blood separating mechanism
Red blood cell blood plasma, the red blood cell why also contained in the blood plasma after separation less than predetermined amount is because red blood cell is flat
Shape, due to the limitation of manufacture craft, intercolumniation cannot accomplish the size less than all red blood cells, therefore have some red blood cells
Can not be by effectively catching.In the controlled range of intercolumniation, the size of predetermined amount specifically can be by the quantity and post of micro-pillar array
Spacing determines that intercolumniation is smaller, predetermined amount is smaller.
As shown in figure 1, in the present embodiment, the second blood separating mechanism 20 includes the second micro-fluidic chip 21, blood plasma returns
Unit 22 and devil liquor recovery unit 23 are received, fluid channel 24 is provided with the second micro-fluidic chip 21.
In the present embodiment, blood plasma recovery unit is specifically referred to for collecting and storing the output of the second blood separating mechanism
The container of the blood plasma of high-purity, devil liquor recovery unit are specifically referred to for collecting and storing blood platelet and red thin less than predetermined amount
The container of born of the same parents, the amount of capacity of container specifically determine by the amount for the blood sample for needing to carry out blood testing, can be according to reality
Border needs to be selected.
In a particular application, fluid channel can be the S-shaped fluid channel of bending, linear pattern fluid channel either other shapes
Fluid channel, in the present embodiment, the shape of fluid channel is not particularly limited.The exemplary S for showing fluid channel to bend in Fig. 1
Shape fluid channel.
As shown in figure 1, the annexation in the whole blood blood plasma piece-rate system 100 that the present embodiment is provided between each part
For：
Blood collection unit 11, at least one first micro-fluidic chip 12 (in Fig. 1 it is merely exemplary show two it is first micro-
Fluidic chip) and two micro-fluidic chips 21 along blood flow direction be arranged in order and by fluid line 30 successively head and the tail connect, blood plasma
Recovery unit 22 is connected by fluid line 30 with the plasma outlet port of the second micro-fluidic chip 21, and devil liquor recovery unit 23 passes through stream
Body pipeline 30 is connected with the waste liquid outlet of the second micro-fluidic chip 21.
In a particular application, fluid line specifically refers to connect between outlet and the entrance for realizing two neighboring part
Connect, so that blood can flow to the universal miniature pipeline of another part from a part, the micro-tube can be specifically glass
Glass micro-pipe, plastic micro, metal micro-tubes, or arbitrarily can transmit blood and not influence the chemistry, physics or life of blood
The micro-tube of thing attribute.
Based on the system architecture shown in Fig. 1, the work of each part in the whole blood blood plasma piece-rate system 100 that the present embodiment is provided
It is as principle：
The blood of blood collection unit collection flows into two the first micro-fluidic chips, two the first micro-fluidic chips point successively
The other haemocyte in blood retains, and obtains the blood plasma of the red blood cell containing blood platelet and less than predetermined amount；Second miniflow
Control chip carries out inertia focusing to blood plasma and obtains the blood plasma, blood platelet and the red blood cell less than predetermined amount of high-purity, high-purity
The blood plasma of degree flows into blood plasma recovery unit, blood platelet and less than predetermined amount red blood cell through flowing into devil liquor recovery unit.
In a particular application, inertia focusing specifically refers to：By inertia focusing come small chi remaining in filtered plasma
When the particles such as very little cell or particle, cell flow in fluid channel, except by main flow driving force flow forward, also in Vertical Square
Wall lift (the Wall Effect that the conduit wall of the shearing force to caused by by the velocity gradient difference as fluid and closure is brought
Lift Force) influence, shearing force and wall lift synthesize inertia force.Under inertia force effect, cell will be in miniflow
Fixed position migration in road, therefore the blood platelet in separated plasma and a small amount of red blood cell are can be used to so as to obtain the blood of high-purity
Slurry.
In one embodiment, plasma outlet port is arranged on above waste liquid outlet.Principle based on inertia force focusing can
Know, the larger blood platelet of quality and a small amount of red blood cell can be arranged by the waste liquid outlet of Action of Gravity Field from below in the blood plasma after separating
Go out, and the blood plasma for being used to carry out the high-purity of blood testing can then flow out from the plasma outlet port of top.
The present embodiment retains son by being set gradually along blood flow direction including the red blood cell for retaining and filtering red blood cell
At least one micro-pillar array of micro-pillar array, and the intercolumniation of each micro-pillar array is differed, make at least one microtrabeculae battle array
The intercolumniation of row is sequentially reduced according to putting in order at least one micro-pillar array, makes blood from the blood separation and culture core
The entrance of piece flows into, and successively by the retention and filtering of at least one micro-pillar array, obtains containing red less than predetermined amount
Cell and hematoblastic blood plasma flow out from the outlet of blood separation and culture chip, it is possible to achieve to minimal amount of blood sample
Quick separating, improve the detection efficiency of blood sample.
As shown in Fig. 2 in one embodiment of the invention, micro-pillar array 13 include along blood flow direction be arranged in order to
A few sub- micro-pillar array, the intercolumniation of every sub- micro-pillar array differ, and the intercolumniation of each sub- micro-pillar array is according at least
One putting in order for sub- micro-pillar array is sequentially reduced, and micro-pillar array 13 is micro- including at least least significant end red blood cell retention is arranged in
Post array 131, red blood cell retain diameter of the intercolumniation more than or equal to red blood cell of sub- micro-pillar array 131.
In a particular application, the quantity of the sub- micro-pillar array included by micro-pillar array can be set according to being actually needed
It is fixed, because the minimum haemocyte of volume in blood is red blood cell, therefore it may only be necessary to ensure that including red blood cell retains sub- micro-pillar array
Can realizes the retention and filtering to all haemocytes in blood.
As shown in Fig. 2 in the present embodiment, micro-pillar array 13 includes retaining along the tumour cell that blood flow direction is arranged in order
Sub- micro-pillar array 134, monocyte retain sub- micro-pillar array 133, leucocyte retains sub- micro-pillar array 132 and red blood cell retention
Micro-pillar array 131；Wherein, tumour cell retains the sub- micro-pillar array 133 of intercolumniation ＞ monocytes retention of sub- micro-pillar array 134
Intercolumniation ＞ leucocytes retain the intercolumniation ＞ red blood cells of sub- micro-pillar array 132 and retain the intercolumniation of sub- micro-pillar array 134.
In a particular application, the quantity of sub- micro-pillar array and species can be configured according to being actually needed.One species
The sub- micro-pillar array of type can set simultaneously it is multiple, with strengthen retain filter effect.
Exemplary when to show micro-pillar array 13 be cylindrical-array, the top view of micro-pillar array 13, arbitrary neighborhood in Fig. 2
The two equal Heterogeneous Permutations of row's microtrabeculae.
In the present embodiment, it is not to face setting that Heterogeneous Permutation, which specifically refers to two adjacent row's microtrabeculaes, but mutually
The certain distance that staggers is set, and whole micro-pillar array is formed an oblique array, the inclined degree of oblique array can be according to reality
Border needs to set.By making the microtrabeculae Heterogeneous Permutation of adjacent row in each micro-pillar array, can improve retention to haemocyte and
Filter effect.
In a particular application, the often row microtrabeculae in micro-pillar array can also face setting.It is if simply every in micro-pillar array
To arrange microtrabeculae all faces to set, form a rectangular array, then haemocyte is easy to along the rectilinear slot outflow do not blocked, from
And reduce the retention to haemocyte and filter effect.
In a particular application, in order to realize preferable haemocyte filter effect, it is necessary to make the intercolumniation of each micro-pillar array
The diameter of the haemocyte of retention and filtering both less than required for it.
In a particular application, the diameter of tumour cell is usually 17 microns~52 microns, and therefore, tumour cell retention is micro-
The intercolumniation of post array should be less than or equal to 17 microns or slightly larger than 17 microns, and most tumour cells are cut with realizing
Stay.For example, the intercolumniation that tumour cell retains sub- micro-pillar array can be in 17 microns~25 micrometer ranges.
In one embodiment, the intercolumniation that tumour cell retains sub- micro-pillar array is 20 microns, and tumour cell retention is micro-
Column diameter is 20 microns.
In a particular application, tumour cell retains the sectional dimension of each microtrabeculae in sub- micro-pillar array and height can basis
Setting is actually needed, for example, height can be more than 52 microns；When tumour cell retains the microtrabeculae width range in sub- micro-pillar array
For 10~30 microns, for example, 10 microns, 15 microns, 20 microns, 25 microns or 30 microns；When microtrabeculae is cylinder, microtrabeculae width
Refer to the diameter of cylinder；When microtrabeculae is square column, microtrabeculae width refers to the square length of side of square column.
In one embodiment, monocyte retains diameter of the intercolumniation less than or equal to monocyte of sub- micro-pillar array
And more than leucocyte, red blood cell and the hematoblastic diameter in blood.
In a particular application, the diameter of monocyte is usually 15 microns~25 microns, and therefore, monocyte retention is micro-
The intercolumniation of post array should be less than or equal to 15 microns or slightly larger than 15 microns, and most monocytes are cut with realizing
Stay.For example, the intercolumniation that monocyte retains sub- micro-pillar array can be in 15 microns~17 micrometer ranges.
In one embodiment, the intercolumniation that monocyte retains sub- micro-pillar array is 15 microns, and monocyte retention is micro-
A diameter of 18 microns of post.
In a particular application, monocyte retains the sectional dimension of each microtrabeculae in sub- micro-pillar array and height can basis
Setting is actually needed, for example, height can be more than 25 microns；When monocyte retains the width model of the microtrabeculae in sub- micro-pillar array
Enclose for 10 microns~30 microns, for example, 10 microns, 15 microns, 20 microns, 25 microns or 30 microns；It is micro- when microtrabeculae is cylinder
Post width refers to the diameter of cylinder；When microtrabeculae is square column, microtrabeculae width refers to the square length of side of square column.
In one embodiment, leucocyte retains the intercolumniation of sub- micro-pillar array and is less than or equal to the diameter of leucocyte and big
Red blood cell and hematoblastic diameter in blood.
In a particular application, the diameter of leucocyte be usually 7 microns~10 microns, 12 microns~20 microns or 14 microns~
20 microns, therefore, the intercolumniation that leucocyte retains sub- micro-pillar array should be less than or equal to 7 microns or slightly larger than 7 microns, with reality
Now to the retention of most leucocytes.For example, the intercolumniation that leucocyte retains sub- micro-pillar array can be at 7 microns~14 microns
In the range of.
In one embodiment, the intercolumniation that leucocyte retains sub- micro-pillar array is 10 microns, leucocyte retention microtrabeculae
A diameter of 12 microns.
In a particular application, leucocyte retains the sectional dimension of each microtrabeculae in sub- micro-pillar array and height can be according to reality
Border needs to set, for example, height can be more than 20 microns；The microtrabeculae width range that leucocyte is retained in sub- micro-pillar array is micro- for 5
Rice~25 microns, for example, 5 microns, 10 microns, 15 microns, 20 microns or 25 microns；When microtrabeculae is cylinder, microtrabeculae width is
Refer to the diameter of cylinder；When microtrabeculae is square column, microtrabeculae width refers to the square length of side of square column.
In a particular application, the diameter of red blood cell be usually 6 microns~8 microns therefore, red blood cell retains sub- micro-pillar array
Intercolumniation should be less than or equal to 6 microns or slightly larger than 6 microns, to realize the retention to most red blood cells.It is for example, red
The intercolumniation of the sub- micro-pillar array of cell retention can be in 4 microns~7 micrometer ranges.
In one embodiment, the intercolumniation that red blood cell retains sub- micro-pillar array is 5 microns, and red blood cell retains the straight of microtrabeculae
Footpath is 10 microns.
In a particular application, red blood cell retains the sectional dimension of each microtrabeculae in sub- micro-pillar array and height can be according to reality
Border needs to set, for example, height can be more than 8 microns；The microtrabeculae width range that red blood cell retains in sub- micro-pillar array is 5 microns
~25 microns, for example, for example, 5 microns, 10 microns, 15 microns, 20 microns or 25 microns；When microtrabeculae is cylinder, microtrabeculae width
Refer to the diameter of cylinder；When microtrabeculae is square column, microtrabeculae width refers to the square length of side of square column.
Based on the structure of the micro-pillar array shown in Fig. 2, the operation principle for the micro-pillar array 13 that the present embodiment is provided is：
Tumour cell retains sub- micro-pillar array and the tumour cell in blood is retained, thin with the tumour in filtering blood
Born of the same parents；Monocyte retains sub- micro-pillar array and the monocyte in blood is retained, with the monocyte in filtering blood；In vain
The sub- micro-pillar array of cell retention retains to the leucocyte in blood, with the leucocyte in filtering blood；
Blood flows into from the entrance of the first micro-fluidic chip, and it is thin to retain sub- micro-pillar array, monokaryon by tumour cell successively
Born of the same parents retain sub- micro-pillar array, leucocyte retains sub- micro-pillar array and red blood cell retains sub- micro-pillar array, thin to the tumour in blood
After born of the same parents, monocyte, leucocyte and red blood cell are retained and filtered, the red blood cell and blood platelet containing less than predetermined amount are obtained
Blood plasma flowed out from the outlet of the first micro-fluidic chip.
In one embodiment, the first blood separating mechanism include being arranged in order along blood flow direction four are first micro-fluidic
Chip, wherein, first micro-fluidic chip includes the sub- micro-pillar array of tumour cell retention, second micro-fluidic chip includes monokaryon
The sub- micro-pillar array of cell retention, the 3rd micro-fluidic chip include leucocyte and retain sub- micro-pillar array, the 4th micro-fluidic chip
Sub- micro-pillar array is retained including red blood cell.
The present embodiment is retained successively by the micro-pillar array including more sub- micro-pillar arrays to the haemocyte in blood
And filtering, it can realize and haemocyte in blood is efficiently separated, obtain the higher blood plasma of purity.
As shown in Figure 3 or Figure 4, in one embodiment of the invention, fluid channel 24 includes the multiple micro- of periodic arrangement
Flow passage unit 241, head and the tail connect multiple micro-channel units 241 successively, and micro-channel unit 241 includes the first semi-circular fluid channel
2411 and the second semi-circular fluid channel 2412, the outlet of the first semi-circular fluid channel 2411 and the second semi-circular fluid channel 2412
Entrance slitless connection.
In a particular application, the quantity of micro-channel unit can be set according to being actually needed, the number of micro-channel unit
Amount is more, better to the separating effect of the blood platelet in blood plasma and the red blood cell of residual, but quantity can excessively reduce separation speed
Degree, so as to reduce blood testing efficiency.Therefore, should consider according to being actually needed to blood platelet and the red blood cell of residual
Separating effect and separating rate requirement, set appropriate number of micro-channel unit.
In one embodiment, the quantitative range of micro-channel unit is 1~50.
In one embodiment, the quantitative range of micro-channel unit is 18.
As shown in figure 3, illustrating for convenience in the present embodiment, exemplary shows that microchannel 24 includes five microchannel lists
Member 241.
In the present embodiment, the external diameter R1 and internal diameter R2 of the first semi-circular fluid channel difference are equal to the first semi-circular fluid channel
Ring cutting the diameter L1, i.e. R1-R2=L1 at upper any place；The external diameter R3 and internal diameter R4 of second semi-circular fluid channel difference are less than second
The maximum ring cutting diameter L2 of semi-circular fluid channel, i.e. R3-R4 ＜ L2；The external diameter R1 of first semi-circular fluid channel is less than the second semi-ring
The internal diameter R4 of shape fluid channel, i.e. R1 ＜ R4；The ring cutting diameter L1 at any place is less than the second semi-circular in first semi-circular fluid channel
The maximum ring cutting diameter L2 of fluid channel, i.e. L1 ＜ L2.
In one embodiment, the ring cutting diameter range of the first semi-circular fluid channel and the second semi-circular fluid channel is 1
Micron~200 microns.
In one embodiment, the maximum ring cutting diameter L2 of the second semi-circular fluid channel is 30 microns.
In one embodiment, the width M1 of the first semi-circular fluid channel is 280 microns, outside the first semi-circular fluid channel
Footpath R1 is 300 microns.
In one embodiment, the width M2 of the second semi-circular fluid channel is 550 microns, outside the second semi-circular fluid channel
Footpath R3 is 980 microns.
The present embodiment is separated by the fluid channel being made up of multiple semi-circular fluid channels to blood plasma, can be efficiently separated
The blood platelet and haemocyte remained in blood plasma, obtains the blood plasma of high-purity, so as to improve the accuracy of blood testing.
As shown in figure 5, in one embodiment of the invention, whole blood blood plasma piece-rate system 100 also includes minitype pneumatic valve
40, the air inlet of minitype pneumatic valve 40 connects constant pressure source of the gas, and the gas outlet of minitype pneumatic valve 40 connects the entrance of blood collection unit 11；
When minitype pneumatic valve 40 is opened, output constant pressure source of the gas promotes blood collection unit to blood collection unit 11 by gas pressure
Blood in 11 flows into the first micro-fluidic chip 12.
In a particular application, minitype pneumatic valve specifically refers to suitable with the volume of the first micro-fluidic chip, for accessing
Opened during gas with constant pressure, and by gas output by gas pressure come the micro valve that drives.The present embodiment
In, constant pressure origin specifically refers to require that the pressure of gas keeps constant in a blood separation process, so that blood can
With by being pushed in the first micro-fluidic chip at the uniform velocity.In different blood separation processes, according to haemoconcentration it is different with
And the requirement to velocity of blood flow, can be according to the pressure size for being actually needed adjustment gas.
The present embodiment promotes blood to enter the first micro-fluidic chip by using minitype pneumatic valve, can both ensure blood
It is not contaminated, can controls the separating rate of blood by controlling pressure and the flow velocity of gas again.
As shown in fig. 6, in one embodiment of the invention, whole blood blood plasma piece-rate system 100, in addition to blood plasma buffer pool
50, the entrance of blood plasma buffer pool 50 is connected by the outlet of fluid line 30 and the first micro-fluidic chip 12 for being arranged in least significant end
Connect, the outlet in blood buffer pond 50 is connected by fluid line 30 with the entrance of the second micro-fluidic chip 21；It is arranged in least significant end
The blood plasma of the first micro-fluidic chip 12 output flow into blood plasma buffer pool 50, blood plasma buffer pool 50 stores to blood plasma, blood plasma
More than blood plasma buffer pool 50 memory capacity when, flow into the second micro-fluidic chip 21.
In the present embodiment, blood plasma buffer pool is specifically referred to for collecting and storing containing for the first blood separating mechanism output
Have blood platelet and less than predetermined amount red blood cell blood plasma container, the amount of capacity of container is specifically by needing to carry out blood testing
The amount of blood sample determine, can be selected according to being actually needed.
The present embodiment is temporarily stored by using blood plasma buffer pool to blood plasma, can in order to control the flow velocity of blood plasma and
The disengaging time of blood plasma.
Filled arrows (→) direction in all embodiments of the invention accompanying drawing represents that blood flows to.
As shown in fig. 7, one embodiment of the present of invention provides a kind of whole blood plasma separation method, it includes：
Step S101：Blood is gathered, the blood includes whole blood blood plasma.
In a particular application, can be adopted by any feasible blood collection mode to gather blood, such as by vacuum
Blood vessel and supporting flexible pipe and syringe needle gather blood, and blood typically refers to the artery or venous blood of human body, naturally it is also possible to
It is animal blood, the mode for gathering blood is not particularly limited in the present embodiment.
In one embodiment, step S101 is performed by the blood collection unit in any of the above-described embodiment.
Step S102：It is more than or equal to the micro-pillar array of the diameter of red blood cell by minimum intercolumniation in the blood
Haemocyte is retained at least once, obtains the blood plasma of the red blood cell containing blood platelet and less than predetermined amount.
In a particular application, haemocyte is retained primarily to filtering out the spectrum point that blood plasma can be disturbed in blood
Analyse the haemocytes such as the leucocyte, monocyte, red blood cell of result.
In one embodiment, step S102 is performed by the first micro-fluidic chip in any of the above-described embodiment, is passed through
Other haemocytes that micro-pillar array is more than red blood cell to the red blood cell in blood and size retain, to realize in blood
The filtering of haemocyte.
In a particular application, the step of being filtered to blood, blood can be entered by same first micro-fluidic chip
Row filters realize at least once, when filtering times more than once when, can also pass through multiple first micro-fluidic chips successively mistake
Hemofiltration liquid, to realize the multiple times of filtration to blood.
Step S103：Inertia focusing is carried out to the blood plasma by the micro-fluidic chip for being provided with fluid channel, enters one
Step filters the blood platelet and the red blood cell less than predetermined amount, obtains the blood plasma of high-purity.
In one embodiment, step S103 is performed by the second micro-fluidic chip in any of the above-described embodiment.
Step S104：Individually reclaim the blood plasma of the high-purity；
Step S105：Reclaim the blood platelet and the red blood cell less than predetermined amount simultaneously.
In a particular application, the blood plasma of high-purity can be separately recovered according to being actually needed using different returnable
And blood platelet and the red blood cell less than predetermined amount, after being acted on due to blood platelet and less than the red blood cell of predetermined amount by inertia force
Mix, therefore, blood platelet and the red blood cell less than predetermined amount are reclaimed simultaneously, the blood plasma of high-purity then by
Individually reclaim for follow-up blood testing.
In one embodiment, step S104 is performed by the blood plasma recovery unit in any of the above-described embodiment, step
S105 is performed by the devil liquor recovery unit in any of the above-described embodiment.
In one embodiment, between the step S102 and S103 in the above method, in addition to step：
The blood plasma is cached.
In the present embodiment, caching specifically refers to enter row buffering and storage to blood plasma, so that blood is passing through of short duration storage
The step of inertia force focuses on is entered back into afterwards.
In a particular application, blood plasma can be cached by arbitrary blood plasma storage container.
In one embodiment, above-mentioned the step of being cached to blood plasma, can specifically be delayed by the blood plasma in above-described embodiment
Pond is rushed to perform.
In a particular application, the whole blood plasma treatment process that embodiment of the method for the invention is provided can be by above-mentioned
Whole blood blood plasma piece-rate system that one system embodiment is provided is realized.
The present embodiment retains the haemocyte in blood by micro-pillar array, and the haemocyte in blood tentatively can be divided
From by carrying out inertia focusing, the further blood platelet and residual red blood cells separated in blood to blood, it is possible to achieve
To haemocyte in minimal amount of blood sample and hematoblastic quick separating, the detection efficiency of blood sample is improved.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement made within refreshing and principle etc., should be included in the scope of the protection.
Claims (10)
- A kind of 1. whole blood blood plasma piece-rate system, it is characterised in that including the first blood separating mechanism and the second blood separating mechanism, First blood separating mechanism includes blood collection unit and at least one first micro-fluidic chip, the first micro-fluidic core Micro-pillar array is provided with piece, the minimum intercolumniation of the micro-pillar array is more than or equal to the diameter of red blood cell, second blood Liquid separating apparatus includes the second micro-fluidic chip, blood plasma recovery unit and devil liquor recovery unit, on second micro-fluidic chip It is provided with fluid channel；The blood collection unit, at least one first micro-fluidic chip and two micro-fluidic chip along blood flow to according to Secondary to arrange and connected from beginning to end successively by fluid line, the blood plasma recovery unit passes through fluid line and the blood of the fluid channel Slurry outlet connection, the devil liquor recovery unit are connected by fluid line with the waste liquid outlet of the fluid channel；The blood of the blood collection unit collection flows at least one first micro-fluidic chip successively, described at least one First micro-fluidic chip retains to the haemocyte in the blood, obtains the red blood cell containing blood platelet and less than predetermined amount Blood plasma；Second micro-fluidic chip blood plasma is carried out inertia focusing obtain the blood plasma of high-purity, blood platelet and Less than the red blood cell of predetermined amount, the blood plasma of the high-purity flows into the blood plasma recovery unit, the blood platelet and less than default The red blood cell of amount is through flowing into the devil liquor recovery unit.
- 2. whole blood blood plasma piece-rate system as claimed in claim 1, it is characterised in that the micro-pillar array includes flowing to along blood At least one sub- micro-pillar array being arranged in order, the intercolumniation of each sub- micro-pillar array differ, each sub- micro-pillar array Intercolumniation be sequentially reduced according at least one putting in order for sub- micro-pillar array, the micro-pillar array comprise at least arrangement Sub- micro-pillar array is retained in least significant end red blood cell, the intercolumniation that the red blood cell retains sub- micro-pillar array is more than or equal to red blood cell Diameter；The red blood cell retains sub- micro-pillar array and the haemocyte in the blood is retained, to filter the blood in the blood Cell.
- 3. the whole blood blood plasma piece-rate system as described in any one of claim 1 or 2, it is characterised in that the micro-pillar array is circle Any of post array, cylindroid array or polygon pillar array, the diameter range of the microtrabeculae in the micro-pillar array are 10 microns~200 microns, 10 microns~200 microns of the altitude range of the microtrabeculae.
- 4. whole blood blood plasma piece-rate system as claimed in claim 1, it is characterised in that the whole blood blood plasma piece-rate system also includes Minitype pneumatic valve, the air inlet of the minitype pneumatic valve connect constant pressure source of the gas, and the gas outlet of the minitype pneumatic valve connects the blood The entrance of collecting unit；When the minitype pneumatic valve is opened, output constant pressure source of the gas to the blood collection unit, promoted by gas pressure described in Blood in blood collection unit flows into first micro-fluidic chip.
- 5. whole blood blood plasma piece-rate system as claimed in claim 1, it is characterised in that the whole blood blood plasma piece-rate system also includes Blood plasma buffer pool, the entrance of the blood plasma buffer pool pass through fluid line and first micro-fluidic chip for being arranged in least significant end Outlet connection, the outlet in the blood buffer pond is connected by fluid line with the entrance of second micro-fluidic chip；The blood plasma for being arranged in first micro-fluidic chip output of least significant end flows into the blood plasma buffer pool, the blood plasma buffering Pond stores to the blood plasma, when the blood plasma exceedes the memory capacity of the blood plasma buffer pool, flows into second miniflow Control chip.
- 6. whole blood blood plasma piece-rate system as claimed in claim 1, it is characterised in that the fluid channel includes periodic arrangement Multiple micro-channel units, the multiple micro-channel unit connect from beginning to end successively, and it is micro- that the micro-channel unit includes the first semi-circular Runner and the second semi-circular fluid channel, the outlet of the first semi-circular fluid channel and the entrance of the second semi-circular fluid channel Slitless connection.
- 7. whole blood blood plasma piece-rate system as claimed in claim 6, it is characterised in that the external diameter of the first semi-circular fluid channel And the difference of internal diameter is equal to the ring cutting diameter at any place in the first semi-circular fluid channel, the second semi-circular fluid channel it is outer The difference of footpath and internal diameter is less than the maximum ring cutting diameter of the second semi-circular fluid channel, the external diameter of the first semi-circular fluid channel Less than the internal diameter of the second semi-circular fluid channel.
- 8. the whole blood blood plasma piece-rate system as described in any one of claim 6 or 7, it is characterised in that the micro-channel unit Quantitative range is 1~50, and the ring cutting diameter range of the first semi-circular fluid channel and the second semi-circular fluid channel is 1 micro- Rice~200 microns.
- 9. whole blood blood plasma piece-rate system as claimed in claim 1, it is characterised in that the plasma outlet port is arranged on the waste liquid Outlet top.
- A kind of 10. whole blood plasma separation method, it is characterised in that including：Blood is gathered, the blood includes whole blood blood plasma；By minimum intercolumniation be more than or equal to red blood cell diameter micro-pillar array the haemocyte in the blood is carried out to It is few once to retain, obtain the blood plasma of the red blood cell containing blood platelet and less than predetermined amount；Inertia focusing is carried out to the blood plasma by the micro-fluidic chip for being provided with fluid channel, it is small further to filter the blood Plate and the red blood cell less than predetermined amount, obtain the blood plasma of high-purity；Individually reclaim the blood plasma of the high-purity；Reclaim the blood platelet and the red blood cell less than predetermined amount simultaneously.
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| CN201710804368.7A CN107702973A (en) | 2017-09-08 | 2017-09-08 | A kind of whole blood blood plasma piece-rate system and method |
| PCT/CN2018/079015 WO2019047498A1 (en) | 2017-09-08 | 2018-03-14 | Whole blood plasma separation system and method |
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