KR20100048507A - Plasma separation device using microfluidic channel and plasma separation method using microfluidic channel - Google Patents

Abstract

PURPOSE: A plasma separation device and a method for manufacturing a plasma separation device are provided, which remove restriction of the shape of the micro channel according to the size of the red blood cell. CONSTITUTION: A method for manufacturing a plasma separation device comprises: a step of cleaning a numbered wafer(200) through the Piranha etch/clean process; a step of applying the negative photo-resist(210) on the upper part of the wafer; a step of soft-baking the coated wafer; a step of exposing using the ultraviolet ray; a step of applying the negative photo-resist to the upper part of the wafer; a step of exposing the ultraviolet ray; a step of developing; a step of injecting the PDMS(230) to the upper part of the wafer; and a step of joining the PDMS and GLASS(240).

Description

Plasma separation device using microfluidic channel and plasma separation method using microfluidic channel

The present invention relates to a plasma separation apparatus using a microchannel and a plasma separation apparatus using a microchannel. The present invention relates to a plasma separation device manufacturing method using microchannels.

In general, the development of micro-electromechanical systems (MEMS) technology has made it possible to fabricate micro scale structures, increasing the focus on microfluidic devices. It is applied to on-a chip and μ-TAS (Micro Total Analysis System). Advantages of such micro-unit devices include rapid response time, reduced reactive volume, reduced use reagent, and reduced energy used by the system, the cost savings, and the ease of mass production.

The system using microfluidic devices is mainly used in bio applications such as biochemical analysis system research, physiological characterization research, micro drug delivery system, etc. among various applications. Current methods in these fields, namely experimental methods on the macro scale, present many inefficient problems such as large sample consumption and long reaction time. And these efforts are underpinning the development of BioMEMS.

In the field of fluid mechanics, blood research is one of the areas of great interest in the field of biotechnology. There are many studies such as the analysis of blood flow in blood vessels in the human body, the field of measurement, and the development of test methods through blood separation and blood analysis.

Blood is largely composed of red blood cells (white blood cells), white blood cells (white blood cells), platelets (blood platelet), blood plasma (blood plasma), which is a liquid component of blood containing red blood cells in it. The composition of plasma is about 90% of water, 7 ~ 8% of plasma protein, and other components include urea, amino acid, uric acid, and other lipids, sugars, inorganic salts, and nonproteinaceous nitrogen compounds. Plasma proteins also contain albumin, globulin, and fibrinogen, which are involved in blood coagulation. Lipids are cholesterol, lecithin and the like. Inorganic salts are sodium, chlorine, potassium, calcium, magnesium and the like, the composition is similar to sea water and plays an important role in maintaining the osmotic pressure in the body to normal. In addition, since the total amount and composition of the plasma change significantly according to the disease, it is used to diagnose the disease and to know the condition of the disease.

Many medical tests are performed using plasma or serum that has had red blood cells removed. Erythrocytes and erythrocyte components have a matrix effect and reduce reproducibility and precision, making standardized testing difficult. This requires a separate method of red blood cells and plasma for more accurate testing. Common methods of separating erythrocytes and plasma include centrifugal separation and a membrane filter. These methods have the disadvantage of requiring a certain amount of blood and time consuming. Plasma separation methods in micro-units include mesh-type filters, comb-shape filters, and diffusion filters. These separation methods also have high flow resistance, low flow rate, There are disadvantages such as large diffusion time. In addition, conventional methods are to separate the plasma and erythrocytes after the blood is collected and to test the separated plasma, which is not suitable for obtaining the test results in real time to determine the patient's condition.

Plasma separation methods require blood collection and testing from patients separately, and require a lot of time for separation and testing after collection. This is a problem that is difficult to use as a test method for detecting the patient's condition in a situation where the patient's condition changes in real time. The patient's condition can be checked in real time even under severe changes. The T-type channel system using the Zweifach-Fung effect has been studied as a real-time plasma separation method using microchannels.

The Zweifach-Fung effect is when the red blood cells in a narrow channel meet branching tubes with different flow rates. Plasma separation method using Zweifach-Fung effect has a disadvantage that must be given a certain flow rate or more with the constraint of the shape of the channel.

In 1968, K.Svanes and B.Zweifach discovered that when branching tubes formed in narrow tubes, red blood cells flowed into branching tubes with low flow resistance and high flow rates. Also, in 1973, Y.C. Fung confirmed that red blood cells flowed in a larger flow direction when the flow ratio differed by more than 2.5: 1 in the same sized branch pipe. This effect is called the Zweifach-Fung effect.

When the diameter of the microtubules is reduced to a size similar to the size of the red blood cells, the red blood cells are aligned at the center of the microtubules. When the microtubules meet branching tubes with different flow ratios, they are flowed into the branching tube with a large flow rate and a low pressure by the pressure difference between both red blood cells. Will enter.

The Zweifach-Fung effect is used as a real-time plasma separation method using microchannels, and a method of separating red blood cells and plasma using a T-type microchannel system has been studied. However, the use of the Zweifach-Fung effect requires the use of narrow microtubules close to the size of red blood cells and a flow rate of more than a certain size. This leads to the limitation of the shape and separation effect of the microtubules.

The present invention has been made to solve the above problems, the shape of the microchannel is not limited to the size of red blood cells, plasma separation apparatus and microchannel using a microchannel that can separate the plasma even if a certain amount of blood does not flow It is to provide a method for manufacturing a plasma separation device using.

The present invention for achieving the object as described above in the plasma separation device for separating blood plasma and red blood cells using a microchannel, the microchannel is a red blood cell having a higher specific gravity than the plasma in the blood flow by gravity It is settled to the lower side, the lower side and the inlet channel is separated into the erythrocyte layer in which the red blood cells are gathered and the plasma layer in which the plasma is collected above the erythrocyte layer; A main channel portion communicating with the inlet channel and into which blood divided into the erythrocyte layer and the plasma layer passes, and a side channel portion communicating with the side surface of the main channel portion and having an inlet formed in the plasma layer to collect the plasma channel; And an outlet channel for adjusting the flow rate ratio of the separation channel by using the flow resistance of the blood passing through the main channel portion, characterized in that the plasma is collected in the subchannel portion.

And, the inlet channel is characterized in that the length of the inlet channel satisfies the formula L = V * h / V _S so that the red blood cells settle and the plasma is collected to the sub-channel part. Where V = blood flow rate, h = distance from the top of the main channel portion to the bottom of the sub channel portion, and V _S = sedimentation rate of red blood cells.

In addition, the inlet channel is characterized in that the length of 5mm or more.

And, the inlet channel is characterized in that the height is 10㎛ or more.

The subchannel unit may be formed in plural.

The main channel portion is wider than the inlet channel.

And, the outlet channel is characterized in that for controlling the flow resistance of the blood in accordance with the length of the outlet channel.

In addition, the first step of cleaning the numbered wafers through a Piranha etch / clean process; Coating a nagative photo-resist 210 onto the wafer; Performing a soft bake on the coated wafer using a direct hot plate; Aligning the photomas on the coated wafer at an aligner and exposing with ultraviolet light; Step 5 of coating the nagative photo-resist (210) to the upper side of the wafer passed through the fourth step; Aligning the photomask on the wafer passed through the five steps and exposing ultraviolet rays; 7 steps of developing by soaking with a developer in the wafer passed through the 6 steps; Injecting PDMS to which the solid agent is added to the upper side of the wafer after the seven steps; Separating the PDMS passed through the eight steps with the wafer, nine steps for contacting the PDMS and GLASS; characterized in that consisting of.

And, the second step and the fifth step is characterized in that the coating height is adjusted using the number of revolutions of the spin coater (spin coater).

In addition, the wafer that passed through the five steps is naturally cooled after the PEB (Post Exposure Bake) in the DHP, characterized in that the six steps are carried out.

Then, the wafer passed through the six steps is characterized in that the seven steps after the natural cooling.

And, the PDMS and GLASS is characterized in that the contact with the plasma asher.

As described above, the red blood cells are settled by gravity and are divided into microchannels divided into an erythrocyte layer and a plasma layer.

First, we established a new technique for real-time plasma separation using the erythrocyte sedimentation effect.

Second, it is applied to real-time plasma separation, lab-on-chip and u-TAS, which is effective for the development of microelements for real-time plasma separation and testing.

Hereinafter, with reference to the accompanying drawings will be described in detail a plasma separation apparatus using a microchannel and a plasma separation apparatus using a microchannel according to the present invention in detail.

1 is a view showing a plasma separation apparatus using a microchannel according to the present invention, Figure 2 is a front view showing a plasma separation apparatus using a microchannel according to the present invention.

As shown in Figure 1 and 2, the microchannel 100 of the present invention, the red blood cells 20 having a higher specific gravity than the plasma 30 during the flow of blood 10 is settled downward by gravity to the red blood cells at the lower side An inlet channel 110 separated from the erythrocyte layer 40 in which the 20 is collected and the plasma layer 50 in which the plasma 30 is collected above the erythrocyte layer 40; The main channel portion 122 is in communication with the inlet channel 110 and the blood 10 divided into the erythrocyte layer 40 and the plasma layer 50 passes, and communicates with the side surface of the main channel portion 122. An inlet is formed in the plasma layer 50, the separation channel 120 consisting of a sub-channel portion 124 where the plasma 30 is collected; The outlet channel 130 for adjusting the flow rate ratio of the separation channel 120 using the flow resistance of the blood 10 passing through the main channel portion 122.

The microchannel 100 is a PDMS channel manufactured using a MEMS process. In addition, the microchannel 100 in the experiment is a mold for replica casting is produced through the SU-8 photoresist (photoresist 210, PR) processing using a photolithography method. Here, SU-8 is a negative photoresist (negative PR), a photosensitive film suitable for manufacturing a high aspect ratio (high aspect ratio) structure.

That is, the microchannel 100 was produced by a replica casting technique using PDMS. PDMS is simpler to process and mass-produced than conventional glass or silicon microchannels. In addition, it has been widely applied to microfluidic devices and lab-on-a-chip for better optical properties due to its transmittance in the 230nm-700nm wavelength band.

Inlet channel 110 according to the present invention is the place where the blood 10 is first flowed by the syringe pump from which the blood 10 is collected, the flow rate of the blood 10 and the length of the inlet channel 110 Accordingly, the red blood cells 20 having a higher specific gravity than the plasma 30 are settled by gravity to form an red blood cell layer 40, and the upper portion of the red blood cell layer 40 has a lower specific gravity than the red blood cells 20. The gathered plasma layer 50 is formed.

Here, the method for calculating the minimum length of the inlet channel 110 for separating the plasma of the present invention is as follows.

The speed at which the red blood cells 20 settle through a fixed space is affected by buoyancy and friction. In other words, when the red blood cells 20 are in the fluid, the gravity acting on the particles due to buoyancy is partially reduced. As the particles move in the fluid, friction forces act against the movement.

3 is a conceptual diagram showing a formula for obtaining the length of the inlet channel according to the present invention, and FIG. 4 is a conceptual diagram showing a table showing the settling depth with respect to the length of the inlet channel.

As shown in Figure 3, the equations shown represent all the forces acting on the particles in the gravitational field, where F _G is the gravity that red blood cells receive, F _B is the buoyancy force that red blood cells receive, and F _D is the frictional force. Indicates. The volume of red blood cells was assumed to be a disc of diameter (D _RC ) of 7.6 μm and the height of red blood cells (h) of 3.5 μm. The density of red blood cells (ρ _C ) was 1100 kg / m ³ , The density (ρ _L ) of plasma was 1030 kg / m ³ . And the viscosity coefficient of plasma was given as 0.0012kg / ms. The settling velocity V _S of erythrocytes calculated based on equations (1), (1-1), (1-2), and (1-3) is 2.54 μm / s.

Here, the length L of the inlet channel 110 is L = V * h / V _S , V = blood flow rate, h = distance from the top of the main channel portion 122 to the bottom of the sub-channel portion , V _S = can be obtained through the settling velocity of red blood cells.

Therefore, in order to collect plasma in the sub-channel part described later, the length L of the inlet channel is preferably 5 mm or more.

As shown in FIG. 4, the settling depth of red blood cells varies according to the flow velocity of various blood. When the table is analyzed, the flow rate is 0.5 μm / min, which is the maximum speed of the flow rate V of blood, and the length of the inlet channel (L ) Should be at least 5 mm so that red blood cells do not interfere with the course of plasma collected into the subchannel. However, when the flow rate V is more than 0.5 μm / min, plasma passes through the subchannels without being collected.

5 is a conceptual diagram illustrating a cell separation apparatus using a microchannel and a cell-free-layer of a microchannel according to the present invention.

As shown in FIG. 5, when the red blood cells move axially in the tube, an outer region of the tube is formed where only the plasma is left. This area becomes wider as the shear rate increases. This area is called the wall effect as it gets closer to the wall. In addition, when the concentration of the red blood cells is high, a crowding effect is applied to hinder the movement of red blood cells from the wall to the center. Based on the above results, there is a region where red blood cells do not exist near the wall in the blood flow, which is called a cell-free-layer (60). Since the cell-free-layer 60 is a region of the wall surface of the channel, the plasma layer may be secured to the upper side of the inlet channel 110 together with the effect of sedimentation of red blood cells.

The reason why the erythrocytes and the plasma are separated from the inlet channel is because of the sedimentation effect of the erythrocytes by gravity as described above, and the cell-free-layer 60 inside the inlet channel. 60) plasma can be separated.

Separation channel 120 according to the present invention is formed in communication with the inlet channel 110, the blood layer of the blood layer 10 divided into the erythrocyte layer 40 and the plasma layer 50 of the plasma layer 50 Allow plasma 30 to be collected separately. The separation channel 120 is in communication with the inlet channel 110 and the blood passing through the main channel portion 122 and the main channel portion 122 through which the blood 10 separated from the inlet channel 110 passes. It consists of the sub-channel part 124 which separates and collects the plasma 30 gathered in the plasma layer 50 among (10). Therefore, the subchannel part 124 is formed to communicate with the side surface of the main channel part 122, so that the plasma 30 of the plasma layer 50 naturally flows in accordance with the flow of the blood 10. To 124.

Preferably, the main channel portion 122 may be wider than the inlet channel 110. This is because the main channel part 122 is wider than the inlet channel 110 so that the red blood cell 20 is forced in the direction opposite to the main channel part 122 by using the rapid dilation tube effect. to be.

Preferably, the subchannel part 124 communicates with the side surface of the main channel part 122, and may be formed above the virtual center line of the main channel part 122. In particular, the subchannel part 124 communicates with the side surface of the main channel part 122, and the distance from the top of the side surface of the main channel part 122 to the bottom end of the subchannel part 124 is the red blood cell 20. When the flow to the main channel portion 122 should be formed so that the red blood cells 20 gathered on the uppermost side does not contact.

Preferably, a plurality of the subchannel portions 124 may be formed by the flow rate ratio of the blood 10. This is because more plasma 30 can be collected as many as the number of the subchannel portions 124 is formed.

In the outlet channel 130 according to the present invention, the blood 10 passing through the main channel part 122 adjusts the flow rate of the separation channel 120 by using the flow resistance. This is because the blood 10 passing through the separation channel 120 is partially plasma 30 is collected in the sub-channel portion 124, the volume ratio of the red blood cells 20 is increased, so that the outlet channel 130 Flow rate ratio of the separation channel 120 can be adjusted using the flow resistance according to the length.

6 is a conceptual diagram showing an equation used for flow rate calculation in a microchannel according to the present invention.

As shown in Figure 6, Q represents the flow rate of each channel, P represents the pressure, V represents the speed. h is the head loss f is the coefficient of friction and D _h is the hydraulic diameter can be calculated using the hydraulic diameter according to the shape of the square channel. The coefficient of friction follows the formula where the Reynolds number is small enough. The assumptions applied in the calculation of the flow rate were calculated assuming that the flow in the channel was a fully developed laminar flow and that the blood was a Newtonian fluid. Where l _eq = length of microchannel, ρ = density of red blood cells.

Based on the above formulas, the flow rate ratio in the main channel portion 122 is 240: 1, and if 20 subchannel portions 124 are made, the total flow rate ratio can be 11: 1, so that more plasma 30 will be collected. Here, of course, the number of the sub-channel unit 124 may be variously performed by those skilled in the art.

Preferably, the outlet channel 130 may adjust the flow resistance according to the length. When the length of the outlet channel 130 increases, the concentrated red blood cells 20 receive a lot of flow resistance, so that the speed of the blood 10 decreases, thereby increasing the ratio of plasma collected to the subchannel part 124.

Next, an embodiment of a plasma separation apparatus using a microchannel according to the present invention will be described.

First, prepare human blood. The blood used herein is heparin-treated human blood and stored at 5 ° C. after blood collection. Heparin is a complex carbohydrate and is widely used for anticoagulant purposes.

Then, the microchannel 100 used in the present invention is prepared. Here, the flow rate is controlled using a syringe pump to form the internal flow of the microchannel 100, and the separation process is optically confirmed using a fluorescence microscope (BX51, Olympus) and a 10bit high speed camera (1200hs, PCO). . The sedimentation effect of the red blood cells 20 is confirmed by measuring from the side of the microchannel 100, and the flow rate in the separation channel 120 is measured using the photographing speed of the high-speed camera in the plane, and the measured flow rate The total amount of plasma separated was measured.

Then, a flow in the microchannel 100 was formed using a syringe pump in the microchannel 100. In this case, the microchannel 100 is injected with the blood 10 in a state where 0.9% physiology saline solution is put. When the blood 10 is injected into the microchannel 100 while the air is filled, air bubbles remain in the separation channel 120 to prevent the flow toward the separation channel 120 having a low flow rate. In order to prevent this, the experiment is performed after removing the air bubbles in the microchannel 100 by injecting physiological saline from all directions of the microchannel 100. At this time, the reason for using 3% physiological saline is to prevent the red blood cells 20 from being deformed by the osmotic effect. The blood 10 has a certain concentration of ions. If the ion concentration is too high or too low, the red blood cells 20 in the blood 10 are osmotic membranes, causing excessive water flow, causing cells to contract (when the external ion concentration is high) or expand (low external ion concentration). When) 0.9% saline was used to prevent cell destruction.

In addition, the speed of the blood 10 is 0.1 μm / min, 0.2 μm / min, 0.3 μm / min, 0.4 μm / min, 0.5 μm / min, and the experiment is repeated.

Next, a method for manufacturing a plasma separation device using a microchannel will be described.

7 is a block diagram showing a method for manufacturing a plasma separation device using a microchannel according to the present invention.

As shown in FIG. 7, the present invention provides a method for cleaning a numbered wafer 200 through a Piranha etch / clean process; Coating a nagative photo-resist (210) (210) onto the wafer (200); Performing a soft bake on the coated wafer 200 using a direct hot plate (DHP); 4 steps of aligning the photomask () above the coated () wafer in the aligner and exposing with ultraviolet light; A five step of coating a nagative photo-resist (210) () on the wafer 200 which has undergone the four steps; Six steps of aligning the photomask 220 to the upper side of the wafer 200 which has undergone the five steps and exposing ultraviolet rays; 7 steps of developing by soaking with a developer in the wafer 200 subjected to the 6 steps; An eight step of injecting the PDMS 230 to which the solid agent is added onto the wafer 200 passed through the seven steps; The PDMS 230, which has passed through the eight steps, is separated from the wafer 200, and the nine steps of contacting the PDMS 230 and the GLASS 240 are performed.

In steps 1 and 2, the numbered wafers 200 are cleaned by Piranha etch / clean (H ₂ SO ₄ & H ₂ O ₂ ) process. Then, the dehydrate bake is dehydrated on the surface of the wafer 200 in a convection oven at 200 ° C. for 30 minutes and then naturally cooled. The negative photo-resist (210) () SU-8 2015, which is prepared in the above process, is coated with an appropriate coating height (10 μm) by controlling the rotation speed of a spin coater. After coating, the relaxation time of the photoresist film is allowed.

In the third step, the wafer 200 coated with the SU-8 photoresist film is soft baked at 65 ° C. for 1 minute and at 95 ° C. for 2 minutes using a direct hot plate (DHP). The purpose of the soft bake is to evaporate the solvent in the photoresist and to improve the bond between the photoresist and the substrate. In addition, the soft bake is divided into two stages because the solvent is evaporated in the first bake, and the second bake is added with a heat treatment concept to improve the aspect ratio. And such a step bake can be expected to effect the thermal stability of the structure, thermal stress reduction. After the baking process, the wafer is cooled at room temperature without rapid thermal changes. This process also involves absorbing moisture into the thin film. Moisture is required for the photochemical reaction in typical DNQ / novolac type photoresist.

In the fourth step, the photoresist-coated wafer 200 and the first mask 220 are aligned in an aligner and exposed using ultraviolet (UV) light. In the case of UV, its wavelength must match the absorption spectrum of the photoresist and the light rays must be parallel. In the experiment, 1000W high-pressure mercury lamp was used, and the intensity of UV was 30mW / cm ² for 365nm. Light uniformity was within ± 5% for the 4 inch region. The exposure energy dose gives 120 mJ / cm ² . At this time, the wafer and the photo mask 220 are in soft contact.

In the fifth step, the SU-8 2050 negative photo-resist 210 is applied evenly on the wafer 200 on which PE is completed, and the spin coater is adjusted to coat the target coating height (80 μm). In this process, air bubbles are easily generated in the photoresist film. This drop may be a problem without disappearing during the coating process and the soft bake, so care must be taken to remove the air bubbles in the photoresist film during spin coating. After that, the wafer is placed on a horizontal position to have a stabilization time. The stabilization time improves the uniformity of the coated photoresist and ensures that it is resistant to the effects of airflow in the convection oven during the soft bake process, so that the appropriate time is allowed. In this experiment, 20 minutes were given. Soft bake in an oven at 65 ° C. for 5 minutes and at 95 ° C. for 15 minutes. After the baking process, cool naturally in a horizontal place.

In step 6, the aligner aligns the photomask 220 according to the conventionally developed photosensitive film, and gives a stabilization time after exposure using ultraviolet rays. The exposure energy amount gives 400 mJ / cm ² based on the 365 nm wavelength band. At this time, the wafer 200 and the photomask 220 are in close contact with each other. The exposure interval at this time is 10 µm.

In the seventh, eighth and ninth steps, the exposed wafer is immersed in a developer and developed. The development is carried out for 4 minutes after developing for 4 minutes, the developer is exchanged for 4 minutes, and then rinsed with Iso Propyl Alcohol (IPA). Then, a solid agent (elastomer) is added to the PDMS 230 in a ratio of 10: 1 and then stirred well. Thereafter, the liquid is poured into the vacuum chamber PDMS 230 to remove the bubble, poured it on a mold (made with SU-8), and then secondary bubble jagger in the vacuum chamber. After that, the oven is solidified at 100 ° C. for 1 hour. Then, the solidified PDMS 230 is carefully removed from the wafer 200, soaked in methanol for a certain time, sonicated, washed again with methanol and dried. The PDMS 230 having the structure formed thereon is cut in units of devices and bonded to the GLASS 240.

Preferably, the wafer 200 that has undergone the five steps is subjected to natural cooling after PEB (Post Exposure Bake) at 65 ° C. for 1 minute and 95 ° C. for 1 minute in DHP. The PEB process is an important process to make the wall of the structure uniform and to withstand the developer.

Preferably, the wafer 200, which has passed through the six steps, is naturally cooled after performing PEB for 2 minutes at 65 ° C and 7 minutes at 95 ° C in an oven before developing.

Preferably, the GLASS 240 and PDMS 230 may be in contact with the plasma asher. This is because this process is an operation requiring sophistication.

As described above, the present invention is a basic technical idea that segregation of plasma by sedimentation of red blood cells in the channel into the Michael channel, which is merely exemplary, and various modifications and equivalents from those skilled in the art will be appreciated. It will be understood that one embodiment is possible, and the true technical protection scope of the present invention should be defined only by the appended claims.

1 is a perspective view showing a plasma separation device using a microchannel according to the present invention.

Figure 2 is a front view showing a plasma separation apparatus using a microchannel according to the present invention.

3 is a conceptual diagram showing a formula for obtaining a length of an inlet channel according to the present invention.

4 is a conceptual diagram showing a table showing the settling depth with respect to the length of the inlet channel.

5 is a conceptual diagram illustrating a cell separation apparatus using a microchannel and a cell-free-layer of a microchannel according to the present invention.

6 is a conceptual diagram showing the equation used for flow rate calculation in a microchannel according to the present invention;

Figure 7 is a block diagram showing a method for manufacturing a plasma separation device using a microchannel according to the present invention.

<Description of the code | symbol about each part of drawing>

10: blood 20: red blood cells

30: plasma 40: erythrocyte layer

50: plasma layer 60: cell-free-layer

100: microchannel 110: inlet channel

120: separate channel 122: main channel portion

124: sub-channel section 130: separate channel

200: wafer 210: resist

220: mask 230: PDMS

240: GLASS

Claims (12)

In the plasma separation device for separating the plasma 30 of the blood 10 and the red blood cells 20 using the microchannel 100, The microchannel 100, The red blood cells 20 having a higher specific gravity than the plasma 30 in the flow of the blood 10 are settled downward by gravity, and the red blood cell layer 40 and the red blood cell layer 40 in which the red blood cells 20 are collected at the lower side thereof. An inlet channel 110 separated into a plasma layer 50 in which plasma 30 is gathered upward; The main channel portion 122 is in communication with the inlet channel 110 and the blood 10 divided into the erythrocyte layer 40 and the plasma layer 50 passes, and communicates with the side surface of the main channel portion 122. An inlet is formed in the plasma layer 50, the separation channel 120 consisting of a sub-channel portion 124 where the plasma 30 is collected; And an outlet channel 130 for adjusting the flow rate ratio of the separation channel 120 by using the flow resistance of the blood 10 passing through the main channel portion 122. Plasma separation device using a micro-channel, characterized in that the plasma channel 30 is collected to the sub-channel portion (124). The method of claim 1, The inlet channel 110, The length of the inlet channel 110 to satisfy the formula L = V * h / V _S so that the red blood cells 20 is settled so that the plasma 30 is collected into the sub-channel unit 124. Plasma separation device using a microchannel. Where V = blood flow rate, h = distance from the top of the main channel portion to the bottom of the sub channel portion, and V _S = sedimentation rate of red blood cells. The method of claim 1, The inlet channel 110, Plasma separation device using a micro-channel, characterized in that the length is more than 5mm. The method of claim 1, The inlet channel 110, Plasma separation device using a micro-channel, characterized in that the height is 10㎛ or more. The method of claim 1, The subchannel unit 124, Plasma separation device using a micro-channel, characterized in that a plurality is formed. The method of claim 1, The main channel unit 122, Plasma separation device using a micro-channel, characterized in that wider than the inlet channel (110). The method of claim 1, The outlet channel 130, Plasma separation device using a micro-channel, characterized in that for controlling the flow resistance of the blood in accordance with the length of the outlet channel. In the plasma separation device manufacturing method using a micro-channel, Cleaning the numbered wafers 200 through a Piranha etch / clean process; Coating a nagative photo-resist 210 onto the wafer 200; Performing a soft bake on the coated wafer 200 using a direct hot plate (DHP); 4 steps of aligning the photomask 220 over the wafer 200 coated by an aligner and exposing using ultraviolet light; A five step of coating a nagative photo-resist 210 onto the wafer 200 which has undergone the four steps;  Six steps of aligning the photomask 220 to the upper side of the wafer 200 which has undergone the five steps and exposing ultraviolet rays; 7 steps of developing by soaking with a developer in the wafer 200 subjected to the 6 steps; An eight step of injecting the PDMS 230 to which the solid agent is added onto the wafer 200 passed through the seven steps; Separating the PDMS (230) after the eight steps with the wafer 200, nine steps to contact the PDMS (230) and GLASS 240; plasma manufacturing apparatus using a micro-channel characterized in that consisting of . The method of claim 8, The second step and the five step, Method of manufacturing a plasma separation device using a micro-channel, characterized in that the coating height is adjusted using the number of revolutions of the spin coater (spin coater). The method of claim 8, The wafer 200 that has passed through the five steps is Method for producing a plasma separation device using a micro-channel characterized in that the six steps after the natural cooling after the PEB (Post Exposure Bake) in DHP. The method of claim 8, The wafer 200 passed through the six steps is Method of producing a plasma separation device using a micro-channel, characterized in that the seven steps after the natural cooling. The method of claim 8, The PDMS 230 and GLASS 240, Method for producing a plasma separation device using a micro-channel, characterized in that the contact with the plasma asher.

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