KR101339118B1 - Apparatus for examining fluids - Google Patents

Abstract

The present invention relates to a fluid test apparatus, and more particularly, to the pre-treatment, reaction and detection of the blood required for the crossmatching test (Lab-on-a-chip) to implement in the channel By using structural features, it is possible to separate blood cells (particles) from blood (fluid), and to mix and react the fluid test apparatus, which allows rapid cross testing.

The fluid test apparatus according to the present invention includes two or more injection parts for injecting a fluid containing microparticles, a separation part for separating particles contained in the fluid injected into the injection part, and a fluid or microparticles separated from the separation part. It includes an intersection formed with a flow path so as to intersect with each other and a reaction part in which the crossed fluid or microparticles react with each other.

The fluid test apparatus according to the present invention having the above configuration implements a cross-compatibility test performed on one chip before blood transfusion, thereby reducing a complicated blood pretreatment process and an increase in test time, and simplifying and speeding up each step. Implemented to achieve, in particular for emergency transfusion patients in a short time, it is possible to quickly determine whether the blood of the donor is suitable blood for the patient.

Lab-on-a-Chip, Cross-Test, Blood Coagulation, ABO, Microfluidic, Channel, Pressure Gradient

Description

Fluid test device {APPARATUS FOR EXAMINING FLUIDS}

The present invention relates to a fluid test apparatus, and relates to a fluid test apparatus that can reduce the time required for cross-compatibility testing by using the structural characteristics of a channel of a lab-on-a-chip.

In general, a crossmatching test is a test to determine whether the blood to be transfused is appropriate for the patient to be transfused, and to determine whether the patient already has an antibody that can react with antigens in the blood. This is done to confirm the blood type and to prevent hemolytic transfusion side effects caused by unexpected antibodies.

Here, the test method (Saline Phase) of the test method is to put about 2 drops of patient serum in the test tube, and then mixed with 1 drop of 2-5% of the blood cell suspension of the donor, and centrifuged for 15 seconds at 3400rpm, Read for aggregation and hemolysis.

Then, in the albumin step, about 1 to 2 drops of 22% of bovine albumin (Bovine Albumin) are mixed in the test tube, incubated at 37 ° C. for 30 minutes, centrifuged at 3400 rpm for about 15 seconds, and the presence or absence of aggregation or hemolysis. Observe.

In addition, in the antiglobulin phase, the test tube is washed three times with saline.After the last washing, saline is completely removed, and after about 1 to 2 drops of AHG (Anti Human Globuline) is added and mixed at about 3400 rpm. After centrifugation for 15 seconds, the presence of aggregation is observed.

In other words, the crossmatching test, which must be performed before receiving a blood transfusion, separates blood from blood donors and blood recipients using a centrifugal separator, and separates blood cells from blood donors (Antigen) and blood donors' plasma ( Antibodies and reagents, including physiological saline, low ionic strength solution (LISS), bovine serum albumin (BSA) polyethylene glycol, and protease (bromelain), are tested.

In addition, a subtest is performed in which the donor's plasma (Antibody), the recipient's blood cell (Antigen), and the reagents according to the test are reacted with each other, and the blood aggregation reaction (Antigen-Antibody binding) in each of the main test and the subtest is performed. By confirming, it is determined whether the transfusion suitability.

In addition, since the aggregation of Antibody in the blood of the recipient and Antigen in the blood cells of the donor is the most important, only the main test may be performed.

However, this cross-compatibility test method is complicated to pretreatment of blood for the test, thereby increasing the test time, especially in emergency transfusion patients with a short time may cause a problem as the time increases.

SUMMARY OF THE INVENTION The present invention has been made to solve the above problems, and the lab-on-a-chip implements the pre-treatment, reaction and detection of blood necessary for the crossmatching test, It is an object of the present invention to provide a fluid test apparatus in which blood cells (particles) are separated from blood (fluid) by using structural features, mixed and reacted with each other, so that a rapid cross test can be performed.

The present invention utilizes at least two or more fluid injectors on one chip, structural features formed in a channel of the fluid injected into the fluid injector, and a separator for separating fluid and particles, and separating the rotor from the separator. A channel is formed on the chip in which the fluid, the intersection intersecting the fluid injected into the other fluid injecting part, and the reaction part in which the two intersecting fluids interact with each other.

As described above, the present invention having the configuration as described above may implement a cross-compatibility test performed before blood transfusion on a single chip, thereby reducing a complicated blood pretreatment process and an increase in test time accordingly. Implemented to simplify and speed up, particularly in the case of emergency transfusion patients, the time can be quickly determined whether the blood of the donor is suitable for the patient.

In order to achieve the above object, the present invention provides two or more injection parts for injecting a fluid containing microparticles into the microfluidic channel; Two or more separators separating the particles contained in the fluid injected into the injection unit; An intersection in which a channel is formed so that the fluid in which particles are separated in the separator intersects; A reaction unit which is a channel formed of a transparent material to mutually react the crossed fluids and to observe the reaction; A pressure unit forming a pressure gradient field such that the fluid inputted into the injection unit is moved to the reaction unit side; .

Preferably, the separator is characterized in that the inclined microstructure having a predetermined inclination with respect to the primary flow direction of the fluid in the upper layer and the lower layer in the microfluidic channel.

Preferably, the inclined fine structure is characterized in that it has a height of a predetermined magnification smaller than the total channel height of the fine channel height.

Preferably, the predetermined magnification height of the inclined microstructure is characterized in that it has a height of 100% to 500% of the size of the fine particles to be separated.

Preferably, the reaction unit is a mixed portion formed of an inclined fine structure having a constant inclination with respect to the primary flow direction of the fluid in the upper layer and the lower layer in the microfluidic channel; And further comprising:

Preferably, the reaction unit is a filter unit for filtering to pass through when the particles contained in the microfluidic aggregates; And further comprising:

Preferably, the filter unit is composed of fine pillars formed at regular intervals.

Preferably, the fine pillars are arranged at intervals of 10% to 30% larger than the size of the particles contained in the microfluid.

Hereinafter, embodiments according to the present invention will be described in detail with reference to the accompanying drawings.

1 is a plan view schematically showing a fluid test apparatus according to the present invention. As shown in the figure, the fluid test apparatus 1 according to the present invention describes only the main test, particularly the test by the physiological saline method and the bromelain method, in the crossmatching test, but is not limited thereto. no.

In addition, the fluid test apparatus 1 according to the present invention includes an injection part 10, a separation part 30, an intersection part 50, a reaction part 70, and a pressure part 90. .

Here, the injection unit 10 is provided to load the blood solution containing the blood and other reagents of the first injection unit 11 and the donor provided with a blood solution containing the blood of the recipient and other reagents The second injection unit 13 is included.

In addition, the separation unit 30 and the first separation unit 31 provided to separate blood cells (particles) in the blood solution (fluid) loaded in the first injection unit 11 of the injection unit 10, And a second separation part 33 provided to separate blood cells (particles) in the blood solution (fluid) loaded in the second injection part 13 of the injection part 10.

In addition, the intersection 50 crosses and reacts the plasma or blood cells separated by the first separator 31 of the separator 30 with the plasma and blood cells separated by the second separator 33. Channels are formed to meet.

In addition, the reaction part 70 is long so that the plasma or blood cells separated from the first separation part 31 and the plasma and blood cells separated from the second separation part 33 react at the intersection 50. It forms a channel, and this part is preferably made transparent so that the user can observe the reaction result.

In addition, the pressure unit 90 provides a pressure to move the blood loaded into the injection unit 10 to the reaction unit 70, the blood is the separation unit 30, the intersection 50 ), A pressure gradient field is formed so as not to reverse the order of the reaction unit 70.

At this time, in the embodiment of the present invention, the first separation unit 31 and the second separation unit 33 may be omitted. The reason is that in the transfusion process, most of the recipients receive only red blood cells in the blood of the donor. This is because the blood of the donor is blood which is separated only from the red blood cell components through a blood donation route or the like, so that blood separation may not be required.

However, since the blood of the recipient is in a state where plasma and blood cells are not separated, at least one separation unit 30 should be provided in the fluid test apparatus 1 according to the embodiment of the present invention.

FIG. 2 is an enlarged projection view of a separator formed in the fluid test apparatus of FIG. 1. FIG. As shown in the figure, the separator 30 of the fluid test apparatus 1 according to the present invention is an inclined microstructure having a height of 1/2 of the height of the channel for aligning and separating particles according to their size. It includes.

Here, the inclined microstructures are alternately formed at the lower and upper portions of the channel, and the lower inclined microstructures 30a and the upper inclined microstructures 30b are formed at an upper portion thereof and are perpendicular to the primary flow direction of the fluid. By forming a secondary pressure gradient in the direction to generate a hydrodynamic interaction between the particles and the structure, the particles are separated.

Accordingly, the blood solution injected from the first injection part 11 and the second injection part 13 passes through the first separation part 31 and the second separation part 33 while in the blood solution (fluid). Allow blood cells (particles) to separate.

In addition, the blood solution loaded in the first injection part 11 and the second injection part 13 applies a negative pressure of the pressure part 90 to obtain a blood solution loaded in the first injection part 10. The first solution 31 is moved to the first separator 31 and the blood solution loaded in the second injection part 13 is moved to the second separator 33.

In addition, in the blood solution of the recipients injected from the first separation unit 31, the plasma, the solution from which the blood cells are separated, is moved in the A direction, and separated from the blood solution of the donor injected from the second separation unit 33, Completed blood cells move in the B direction, and the moved blood cells of the donor and the blood plasma of the recipients pass through the intersection 50.

Then, while crossing the cross section 50 and cross (cross) and mixed with each other, which corresponds to the main test of the cross-compatibility test, the blood cells and plasma passing through the cross section 50 of the reaction section 70 They interact with each other over long channels.

Here, by adjusting the pressure of the pressure unit 90, the blood cells and plasma passing through the cross section 50 is prevented from entering the pressure unit 90 out of the reaction unit 70, the reaction unit 70 In the case where the donor's blood is not suitable for transfusion to the recipient, the antigen in the blood cell of the donor and the antibody in the plasma of the donor undergo an anti-antigen-antibody binding.

Accordingly, the reaction unit 70 starts to coagulate, and the reaction unit 70 is transparent so that an observer can identify the reaction from the outside so that the coagulated blood can be visually identified by the antigen-antibody binding. It is preferable to be manufactured using a material.

In the case where blood is coagulated in the reaction unit 70, the blood of the donor is unsuitable for the recipient, so that it is not transfused to the recipient.

On the other hand, in the cross-fit test, in the case of the physiological saline method, the blood loaded into the first injection section 11 and the second injection section 13 is the ratio of the plasma amount of blood of the recipient and the amount of blood blood cells of the donor It is adjusted to be about 2: 1, the blood cells of the donor blood is based on the saline blood cell suspension of about 3 to 5%.

In addition, in the bromelain method, which is a protein enzymatic method, the blood loaded in the first injection unit 11 and the second injection unit 13 includes blood plasma volume of blood of the recipient, blood cell volume of the blood of the donor, and bromelain solution ( 0.5%) is adjusted to about 2: 1: 1, but the blood cells of the donor blood are based on the physiological saline cell suspension of about 3 to 5%.

In addition, when the blood and bromelain solution is moved to the reaction unit 70, after reacting for about 15 minutes at about 37 ℃, to determine whether the agglomeration, bromelain solution is a proteolytic element, part of the blood cell membrane structure By peeling off, the antigen is exposed to the surface and sialic acid, a negatively charged cell structure, is removed.

Here, the blood cells are negatively charged by the carboxyl group of the sialic acid (Carboxylic Group) present in the surface membrane, by removing the sialic acid, thereby lowering the zeta potential of the blood cell membrane, thereby lowering the repulsive force between the blood cells, Access between blood cells is facilitated, allowing aggregation reactions to occur better.

3 is a plan view schematically showing an embodiment including a mixing unit in the fluid test apparatus of FIG. 1. As shown in the figure, the fluid test apparatus 1 according to the present invention further includes a mixing unit 71 in the fluid test apparatus 1 of FIG.

In addition, the structure of the mixing part 71 has an inclined structure having a constant inclination with respect to the primary flow direction of the fluid in only one of the upper layer and the lower layer of the channel, and thus is perpendicular to the primary flow direction of the fluid. A secondary gradient pressure field is formed in the direction, causing the particles to mix, thereby creating a hydrodynamic interaction between the particles in the fluid and the structure.

In addition, the plasma or blood cells separated in the first separation unit 31 and the blood cells or plasma separated in the second separation unit 33 pass through the cross section 50 to react the cross-crossed blood ( 70).

Therefore, while passing through the mixing portion 71, the antigen in the blood cells and the antibody in the serum are mixed with each other, thereby increasing the possibility of binding due to the collision.

4 is a plan view schematically showing an embodiment including a filter part including a micropillar in the fluid test apparatus of FIG. 1, and FIG. 5 is a cross-sectional view of the filter part of FIG. 1. As shown in the figure, the fluid test apparatus 1 according to the present invention

As shown in the figure, the fluid test apparatus 1 according to the present invention further includes a filter unit 73 in the reaction unit 70.

In addition, the structure of the filter unit 73 arranges the micropillars 73a at regular intervals. For example, considering about 7 μm of the size of red blood cells, about 10 μm, one red blood cell may pass. Aggregated and enlarged red blood cells are arranged at intervals that cannot pass.

In addition, the plasma or blood cells separated in the first separation unit 31 and the blood cells or plasma separated in the second separation unit 33 pass through the fluid cross-section 30 to react the cross-crossed blood. Moving to 70, the antigen in the blood cells and the antibody in the serum form an antigen-antibody bond with each other while passing through the filter unit 73.

Then, when coagulation between blood cells occurs, blood cells coagulated between the micropillars 73a do not pass through the filter portion 73 and are filtered, thereby preventing the progress of the blood passing therethrough.

Accordingly, when the blood passing through the filter unit 73 does not proceed anymore, since antigen-antibody binding occurs and the blood coagulates, it is determined that the blood of the donor is inadequate for the recipient and the blood of the donor. Do not give blood to the recipient.

FIG. 6 is a plan view schematically illustrating an embodiment further including a third injector in the fluid test apparatus of FIG. 1. As shown in the figure, the fluid test apparatus according to the present invention further includes a third injection unit 15.

And, the plasma or blood cells separated in the first separation unit 31, the blood cells or plasma separated in the second separation unit 33, and the specific reagent injected into the third separation unit 35 cross-section After 50, the blood crosses each other and moves to the reaction unit 70 for reacting the specific reagent, and the reaction is performed.

Here, the specific reagent is preferably used as a physiological saline solution, low ion strength solution (LISS), bovine serum albumin (BSA), polyethylene glycol, protease (bromelain) and the like.

In the above described exemplary embodiments of the present invention by way of example, the scope of the present invention is not limited only to this specific embodiment, and those skilled in the art within the scope described in the claims of the present invention Changes may be made as appropriate.

1 is a plan view schematically showing a fluid test apparatus according to the present invention;

FIG. 2 is an enlarged projection view of a separator formed in the fluid test apparatus of FIG. 1. FIG.

3 is a plan view schematically showing an embodiment including a mixing unit in the fluid test apparatus of FIG.

4 is a plan view schematically showing an embodiment including a filter unit including a micropillar in the fluid test apparatus of FIG. 1.

5 is a cross-sectional view of the filter unit of FIG. 1.

FIG. 6 is a plan view schematically showing an embodiment further including a third injector in the fluid test apparatus of FIG. 1; FIG.

      <Brief description of reference numerals for the main parts of the drawings>

1: fluid test device 10: injection unit

11: first injection part 13: second injection part

15: third injection unit 30: separation unit

31: first separator 33: second separator

50: intersection 70: reaction part

71: mixing section 73: filter section

73a: micropillar 90: pressure part

Claims (10)

Two or more injection parts for injecting a fluid containing microparticles; Separation unit for separating the particles contained in the fluid injected into the injection unit; An intersection where a flow path is formed such that the fluid or microparticles separated in the separation part cross each other; And It includes a reaction unit in which the intersected fluid or microparticles react with each other, And the reaction part includes a filter part to prevent a reactant produced by the reaction from passing through. The method according to claim 1, And a pressure unit providing pressure to move the fluid injected into the injection unit to the reaction unit. The method according to claim 1, The separation unit fluid test apparatus, characterized in that the inclined microstructure having a constant inclination with respect to the primary flow direction of the fluid in the upper or lower layer of the micro-channel. The method of claim 3, The height of the inclined fine structure is a fluid test device, characterized in that smaller than the height of the fine flow path by a certain magnification. The method according to claim 1, The reaction unit further comprises a mixing unit for mixing the fluid or fine particles separated in the separation unit. The method of claim 5, The mixing unit further comprises an inclined microstructure having a constant inclination with respect to the primary flow direction of the fluid in the upper or lower layer of the microchannel. delete The method according to claim 1, The filter unit further comprises a micropillar on the upper or lower portion of the micro channel. The method according to claim 1, The reaction unit further comprises a window of a transparent material that can determine whether the reaction of the separated fluid or microparticles fluid. The method according to claim 1, And the fluid is blood.

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