CN114509323A - Centrifugal micro-fluidic whole blood separation plasma structure - Google Patents
Centrifugal micro-fluidic whole blood separation plasma structure Download PDFInfo
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- CN114509323A CN114509323A CN202210177033.8A CN202210177033A CN114509323A CN 114509323 A CN114509323 A CN 114509323A CN 202210177033 A CN202210177033 A CN 202210177033A CN 114509323 A CN114509323 A CN 114509323A
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- 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
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
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- 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
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
- G01N2035/1027—General features of the devices
- G01N2035/1034—Transferring microquantities of liquid
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Abstract
The invention discloses a centrifugal microfluidic whole blood plasma separation structure. This centrifugal micro-fluidic whole blood separation plasma structure includes: the whole blood plasma separation device comprises a chip substrate, wherein a plurality of whole blood plasma separation modules are arranged on the chip substrate around a central point; the whole blood separation plasma module includes: the device comprises a sample adding groove, a plasma quantifying groove, a red blood cell separating hole, a waste liquid groove and a plasma extracting groove; the sample adding groove is communicated with the plasma quantifying groove through the liquid inlet flow channel, the plasma quantifying groove is communicated with the red blood cell separation hole, the plasma quantifying groove is communicated with the waste liquid groove through the overflow flow channel, and the plasma quantifying groove is connected with the plasma extraction groove through the siphon flow channel. And (3) centrifuging for the first time, so that the whole blood sample in the sample adding groove enters the plasma quantifying groove and the red blood cell separating hole and is filled with the whole blood sample, and the redundant whole blood sample enters the waste liquid groove. After standing, red blood cells and plasma are separated. After the secondary centrifugation, the plasma in the plasma quantifying groove completely enters the plasma extracting groove, and a plurality of whole blood samples can be separated simultaneously, so that the efficiency of whole blood separation is greatly improved.
Description
Technical Field
The invention relates to the field of microfluidic chips, in particular to a centrifugal microfluidic whole blood plasma separation structure.
Background
Blood testing is the most common testing method in biomedicine, clinical diagnosis and health quarantine diagnosis, and in blood testing, plasma is separated from whole blood drawn from a patient, which is also an essential part of the blood testing process. At present, the whole blood separation method adopted by most hospitals and laboratories is realized by a separation gel procoagulant blood collection tube, namely low-speed centrifugation is used, so that red blood cells and serum are layered to two sides of the separation gel due to density difference; the blood collecting tube is provided with coagulation promoting components and separating gel, and the coagulation promoting components can promote the coagulation process, so that erythrocytes and blood coagulation proteins are aggregated and are easy to centrifuge down. However, such a whole blood separation method generally has the technical problems of slow separation speed and low efficiency.
The large-scale centrifuge is adopted to separate the serum and the blood cells, so that a large amount of waste of samples is easily caused; and because the centrifugal device is bulky, the mechanical structure is complicated, and the serum separation process and the detection are separately and independently carried out, the loss and the pollution of blood samples are easily caused, and the detection steps are more complicated.
Therefore, the whole blood separation by the prior art has the technical problems of complicated separation steps and low separation efficiency.
Disclosure of Invention
In view of the above, the main object of the present invention is to provide a centrifugal microfluidic whole blood plasma separation structure with few whole blood separation steps and high separation efficiency.
In order to achieve the purpose, the technical scheme of the invention is realized as follows:
this centrifugal micro-fluidic whole blood separation plasma structure includes: the device comprises a chip base body and a chip cover plate, and is characterized in that a plurality of whole blood plasma separation modules are arranged on the chip base body around a central point; the whole blood plasma separation module comprises: the device comprises a sample adding groove, a plasma quantifying groove, a red blood cell separating hole, a waste liquid groove and a plasma extracting groove;
the sample adding groove is communicated with the plasma quantifying groove through a liquid inlet flow channel, the plasma quantifying groove is communicated with the red blood cell separation hole, the plasma quantifying groove is communicated with the waste liquid groove through an overflow flow channel, and the plasma quantifying groove is connected with the plasma extracting groove through a siphon flow channel.
In one embodiment, the chip substrate is circular, and a fixing device connected with a rotating mechanism for driving the chip substrate to rotate is arranged at the center of the chip substrate.
In one embodiment, a sample adding hole is arranged beside the sample adding groove, and the sample adding hole is communicated with the sample adding groove.
In one embodiment, the waste liquid tank and the plasma quantifying tank are respectively provided with an air hole, and the waste liquid tank and the plasma quantifying tank are respectively communicated with the air hole through a flow passage.
In one embodiment, the width of the flow channel is 0.1-0.5mm, the depth of the flow channel is 0.1-0.5mm, and the surface of the siphon flow channel is subjected to hydrophilic modification.
In one embodiment, the volume ratio of the plasma quantification tank to the red blood cell separation hole is 1: 2.
The centrifugal microfluidic whole blood separation plasma structure has the following beneficial effects:
this centrifugal micro-fluidic whole blood separation plasma structure includes: the chip comprises a chip base body and a chip cover plate, and is characterized in that a plurality of whole blood plasma separation modules are arranged on the chip base body around a central point; the whole blood separation plasma module includes: the device comprises a sample adding groove, a plasma quantifying groove, a red blood cell separating hole, a waste liquid groove and a plasma extracting groove; the sample adding groove is communicated with the plasma quantifying groove through the liquid inlet flow channel, the plasma quantifying groove is communicated with the red blood cell separation hole, the plasma quantifying groove is communicated with the waste liquid groove through the overflow flow channel, and the plasma quantifying groove is connected with the plasma extraction groove through the siphon flow channel. Injecting a whole blood sample into the sample adding groove, and starting a first centrifugal operation; the whole blood sample in the sample adding groove enters the plasma quantifying groove and the red blood cell separation hole through the liquid inlet flow channel, and after the plasma quantifying groove and the red blood cell separation hole are filled, redundant samples enter the waste liquid groove through the overflow channel. After the first centrifugation operation is completed, red blood cells and plasma in the whole blood sample start to be separated, and the plasma separated from the whole blood sample is quantified by the volume in the plasma quantifying tank.
After the siphon flow channel is filled with plasma, a second centrifugal operation is carried out, the plasma in the plasma quantifying groove completely enters the plasma extracting groove, red blood cells in the red blood cell separating hole are completely remained in the hole, and the plasma in the plasma extracting groove can be used for a subsequent analysis and detection test. The centrifugal microfluidic whole blood separation structure has the advantages of few operation steps and simple structure. And because a plurality of whole blood separation plasma modules are arranged on the chip substrate around the central point, the separation operation of a plurality of plasma samples can be simultaneously carried out, the whole blood separation efficiency is greatly improved, and the time required by blood detection is reduced.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the prior art descriptions will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic diagram of a centrifugal microfluidic whole blood plasma separation configuration according to an embodiment of the present disclosure;
FIG. 2 is a schematic diagram of a whole blood plasma separation module after a first centrifugation according to one embodiment of the present disclosure;
FIG. 3 is a schematic diagram of a whole blood plasma separation module after a second centrifugation according to one embodiment of the present disclosure.
[ description of main reference symbols ]
1. A chip substrate; 2. a whole blood plasma separation module; 21. a sample adding slot; 22. a plasma dosing device; 23. a red blood cell separation well; 24. a waste liquid tank; 25. a plasma extraction tank; 3. a siphon runner; 4. a fixing device; 5. and (4) sample adding holes.
Detailed Description
The centrifugal microfluidic whole blood plasma separation structure of the invention is further described in detail with reference to the accompanying drawings and embodiments of the invention.
It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict. The present invention will be described in detail below with reference to the embodiments with reference to the attached drawings.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
It should be noted that the terms "first," "second," and the like in the description and claims of this application and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the application described herein are, for example, capable of operation in sequences other than those illustrated or otherwise described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
Spatially relative terms, such as "above … …," "above … …," "above … … surface," "above," and the like, may be used herein for ease of description to describe one device or feature's spatial relationship to another device or feature as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if a device in the figures is turned over, devices described as "above" or "on" other devices or configurations would then be oriented "below" or "under" the other devices or configurations. Thus, the exemplary term "above … …" can include both an orientation of "above … …" and "below … …". The device may be otherwise variously oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly.
As shown in fig. 1 to 3, the centrifugal microfluidic whole blood separation plasma structure includes: the whole blood plasma separating device comprises a chip substrate 1, wherein a plurality of whole blood plasma separating modules 2 are arranged around a central point on the chip substrate 1; the whole blood separation plasma module 2 includes: a sample adding groove 21, a plasma quantifying groove 22, a red blood cell separating hole 23, a waste liquid groove 24 and a plasma extracting groove 25;
in one embodiment, the depth of the plasma quantifying groove 22 is set to be equal to or greater than 0.5mm, so that the inaccurate quantifying result caused by the generation of bubbles in the plasma quantifying groove 22 due to too shallow liquid layer when the depth of the plasma quantifying groove 22 is less than 0.5mm is avoided. In one embodiment, there is a step in the waste liquid tank 24 with a height greater than 1mm, the presence of which prevents the waste liquid from flowing back to the plasma quantification tank 22.
The sample adding groove 21 is communicated with the plasma quantifying groove 22 through a liquid inlet flow channel (preferably, the width and the depth of the liquid inlet flow channel are less than or equal to 0.5mm, and the width and the depth of the liquid inlet flow channel are prevented from being too large, when a plasma sample enters the plasma quantifying groove 22 and the red blood cell separation hole 23, the flow rate of the plasma sample is too high, so that bubbles are formed in the plasma quantifying groove 22 and the red blood cell separation groove 23), the plasma quantifying groove 22 is communicated with the red blood cell separation hole 23, the plasma quantifying groove 22 is communicated with the waste liquid groove 24 through an overflow flow channel (preferably, the depth of the overflow flow channel is greater than or equal to 0.5mm, the depth of the overflow flow channel is less than that of the plasma quantifying groove 22, and the depth difference between the depth of the overflow flow channel and the depth of the plasma quantifying groove 22 is not greater than 1 mm), and the plasma quantifying groove 22 is connected with the plasma extracting groove 25 through the siphon flow channel 3.
The centrifugal microfluidic whole blood separation plasma structure is externally connected with a centrifugal driving device, a whole blood sample in a sample adding groove 21 enters a plasma quantifying groove 22 and a red blood cell separation hole 23 in a rotating centrifugal mode and is filled with the whole blood sample, and an excess whole blood sample enters a waste liquid groove 24. After standing for a certain period of time, the red blood cells in the whole blood sample enter the red blood cell separation hole 23, and the plasma is in the plasma quantifying groove 22. After the centrifugation, the whole plasma in the plasma quantifying tank 22 enters the plasma extracting tank 25 for the subsequent plasma detection.
The centrifugal microfluidic whole blood separation structure has the advantages of few operation steps and simple structure. And because a plurality of whole blood separation plasma modules 2 are arranged around the central point on the chip substrate 1, the separation operation of a plurality of whole blood samples can be simultaneously carried out, the efficiency of whole blood separation is greatly improved, and the time required by blood detection is reduced.
In order to facilitate the rotation of the chip base 1, the chip base 1 is made circular, and a fixing device 4 for connecting with a rotation mechanism for driving the chip base 1 to rotate is arranged at the center of the chip base 1. The fixing device 4 is externally connected with centrifugal driving equipment, so that the chip matrix 1 can rotate around the center of the chip matrix to complete centrifugal operation.
In order to facilitate the injection of the whole blood sample into the sample addition well 21, a sample addition hole 5 is formed beside the sample addition well 21, and the gas addition hole 5 is communicated with the sample addition well 21.
In order to extract the redundant whole blood sample and plasma, air holes are respectively formed beside the waste liquid tank 24 and the plasma quantifying tank 25, so that the waste liquid tank 24 and the plasma quantifying tank 25 are respectively communicated with the air holes through flow passages. In one embodiment, the diameter of the air hole is larger than 1mm, and the cross-sectional area of the air hole is larger than the cross-sectional areas of the siphon flow channel 3 and the overflow flow channel, so that the air pressure inside the chip is kept stable when the centrifugal microfluidic whole blood separation plasma structure is centrifuged.
In one embodiment, the siphon flow channel 3 has a width of 0.1-0.5mm and a depth of 0.1-0.5mm, and the surface of the siphon flow channel 3 is modified by hydrophilicity, so that the whole blood sample after the first centrifugation operation can not enter the plasma extraction tank 25 in advance, thereby avoiding affecting the accuracy of blood detection. After the first centrifugation is finished, standing for a period of time, and filling the siphon flow channel 3 with plasma due to capillary action; the plasma in the plasma quantifying tank 22 is again centrifuged and enters the plasma extraction tank 25 through the siphon flow path 3, thereby completing the extraction of the plasma.
In order to facilitate the separation of plasma, the volume ratio of the plasma quantifying tank 22 to the red blood cell separating hole 23 is set to 1: 2. In order to ensure that all of the plasma is present in the plasma quantifying tank 22, the volume of the plasma quantifying tank 22 is made slightly smaller than the volume of the plasma in the whole blood sample in the plasma quantifying tank 22 and the red blood cell separation hole 23.
The centrifugal microfluidic whole blood separation plasma structure can be used for separating plasma from whole blood and can also meet the requirements of separating precipitates and extracting supernatant. In a specific embodiment, the invention, when in use, has the following working flow:
s1, injecting the whole blood sample into the sample injection slot 21 through the sample injection hole 5, operating the driving device to make the chip matrix 1 perform the first centrifugation, setting the rotating speed at 3000 and 5000rpm, and the centrifugation time at 90-300 seconds.
The whole blood sample in the sample adding groove 21 enters the plasma quantifying groove 22 and the red blood cell separation hole 23 through the liquid inlet flow channel, and after the plasma quantifying groove 22 and the red blood cell separation hole 23 are filled with the plasma sample, the redundant plasma sample enters the waste liquid groove 24 through the overflow channel.
S2, after the whole blood sample in the plasma quantifying tank 22 and the red blood cell separation hole 23 is centrifuged for a certain period of time, red blood cells and plasma in the whole blood sample start to be separated, red blood cells are precipitated in the red blood cell separation hole 23, and plasma is in the plasma quantifying tank 22.
S3, the plasma separated from the whole blood sample is quantified by the volume of the plasma quantifying tank 22.
S4, stopping the centrifugation for 10-30 seconds after the chip substrate 1 is centrifuged for the first time, and waiting for the siphon flow channel 3 to be filled with the plasma.
S5, operating the driving device to make the chip matrix 1 to perform the second centrifugation, wherein the centrifugation speed is 3000-.
The plasma in the plasma quantifying groove 22 completely enters the plasma extracting groove 25, the red blood cells in the red blood cell separating hole 23 are completely remained in the hole, and the plasma in the plasma extracting groove 25 can be used for a subsequent analysis and detection test to finish the second centrifugal operation.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention.
Claims (6)
1. A centrifugal microfluidic whole blood separation plasma structure comprising: the chip substrate (1) is characterized in that a plurality of whole blood separation plasma modules (2) are arranged on the chip substrate (1) around a central point; the whole blood-plasma separation module (2) comprises: a sample adding groove (21), a plasma quantifying groove (22), a red blood cell separating hole (23), a waste liquid groove (24) and a plasma extracting groove (25);
the sample adding groove (21) is communicated with the plasma quantifying groove (22) through a liquid inlet flow channel, the plasma quantifying groove (22) is communicated with the red blood cell separation hole (23), the plasma quantifying groove (22) is communicated with the waste liquid groove (24) through an overflow flow channel, and the plasma quantifying groove (22) is connected with the plasma extracting groove (25) through a siphon flow channel (3).
2. A centrifugal microfluidic whole blood plasma separation structure according to claim 1, wherein the chip base body (1) is circular, and a fixing device (4) for connecting with a rotating mechanism for driving the chip base body (1) to rotate is arranged at the center of the chip base body (1).
3. A centrifugal microfluidic whole blood plasma separation structure according to claim 1, wherein the sample adding groove (21) is provided with sample adding holes (5) beside, and the sample adding holes (5) are communicated with the sample adding groove (21).
4. The centrifugal microfluidic whole blood plasma separation structure according to claim 1, wherein air holes are formed beside the waste liquid tank (24) and the plasma quantification tank (25), and the waste liquid tank (24) and the plasma quantification tank (25) are respectively communicated with the air holes through flow channels.
5. The centrifugal microfluidic whole blood plasma separation structure according to claim 1, wherein the width of the siphon flow channel (3) is 0.1-0.5mm, the depth is 0.1-0.5mm, and the surface of the siphon flow channel (3) is hydrophilic modified.
6. A centrifugal microfluidic whole blood separation plasma structure according to claim 1, characterized in that the volume ratio of the plasma quantification groove (22) and the red blood cell separation hole (23) is 1: 2.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210177033.8A CN114509323A (en) | 2022-02-24 | 2022-02-24 | Centrifugal micro-fluidic whole blood separation plasma structure |
| PCT/CN2022/108945 WO2023159871A1 (en) | 2022-02-24 | 2022-07-29 | Centrifugal microfluidic structure for separating plasma from whole blood |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210177033.8A CN114509323A (en) | 2022-02-24 | 2022-02-24 | Centrifugal micro-fluidic whole blood separation plasma structure |
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| CN114509323A true CN114509323A (en) | 2022-05-17 |
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| CN202210177033.8A Pending CN114509323A (en) | 2022-02-24 | 2022-02-24 | Centrifugal micro-fluidic whole blood separation plasma structure |
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| CN (1) | CN114509323A (en) |
| WO (1) | WO2023159871A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2023159871A1 (en) * | 2022-02-24 | 2023-08-31 | 含光微纳科技(太仓)有限公司 | Centrifugal microfluidic structure for separating plasma from whole blood |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114509323A (en) * | 2022-02-24 | 2022-05-17 | 含光微纳科技(太仓)有限公司 | Centrifugal micro-fluidic whole blood separation plasma structure |
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2022
- 2022-02-24 CN CN202210177033.8A patent/CN114509323A/en active Pending
- 2022-07-29 WO PCT/CN2022/108945 patent/WO2023159871A1/en unknown
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| JP2009150733A (en) * | 2007-12-20 | 2009-07-09 | Panasonic Corp | Living body analyzing device, and blood separation method using it |
| CN204544220U (en) * | 2015-03-30 | 2015-08-12 | 博奥生物集团有限公司 | A kind of rotating centrifugal separation of whole blood chip |
| JP2018163169A (en) * | 2018-06-26 | 2018-10-18 | シスメックス株式会社 | Cartridge for measurement and liquid feeding method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2023159871A1 (en) * | 2022-02-24 | 2023-08-31 | 含光微纳科技(太仓)有限公司 | Centrifugal microfluidic structure for separating plasma from whole blood |
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| WO2023159871A1 (en) | 2023-08-31 |
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