JP4844318B2 - Microchannel device - Google Patents

Description

  The present invention relates to a microchannel device having a blood filtration function, and more specifically, clinical tests in the fields of chemistry, biochemistry, medicine, etc., particularly blood separation for use in point-of-care testing or home use is necessary. The present invention relates to a microchannel device used for measurement.

  Biochemical tests that measure components in blood are widely used for various diagnoses and observations, and are important tests as clinical tests. Various biochemical testing devices have been developed, and many samples and many test items are analyzed. Conventional health examinations and diagnosis of disease states have been carried out by taking a large amount of blood of several cc from a patient and analyzing it from measurements obtained with a large-scale automatic blood analyzer. Usually, such an automatic analyzer is installed in a medical institution such as a hospital, has a large scale, and its operation is limited to those who have expertise.

  However, in recent years, by applying the microfabrication technology used for the production of semiconductor devices that have advanced extremely recently, analyzers such as various sensors are arranged on a substrate of several millimeters to several centimeters square, and the blood of the subject or the like is placed there. The development and practical application of new devices that can guide body fluids and instantly grasp the health status of subjects are increasing. With the advent of such low-cost devices, health insurance benefits can continue to be reduced by enabling daily health management of the elderly at home in an aging society that should come. Also, in the field of emergency medicine, various social effects are expected, such as being able to take appropriate measures if this device can be used to quickly determine the presence or absence of infection (such as hepatitis or acquired immune deficiency). This is a technical field that is attracting a great deal of attention. As described above, instead of the conventional automatic analyzer, a small and simple blood analysis method and a blood analyzer aiming to carry out blood analysis by one's own hands in each home have been developed. In such a test, it is desired to remove a blood cell component in a sample because a specific component in blood, particularly a blood cell component, causes a high background in the measured value or interferes with the performance of the measuring device. Further, in many cases, it is desirable that the inspection apparatus can measure with a small amount of sample.

  In conventional health examinations and diagnosis of disease states, a method using centrifugation has been used to remove blood components in the sample. However, several cc of blood is required, and the supernatant is removed after centrifugation. However, skillful work such as sucking out was necessary.

  The blood cell component separation structure disclosed in Patent Document 1 discloses a chip that separates blood by centrifugal force, but after introducing blood as a specimen and separating it by centrifugal force, the chip is rotated by 90 ° for separation. This is a complicated procedure that weighs and feeds the collected blood and rotates it 90 ° to mix it with the reagent. Removing the microchip takes time, and the microchip is dropped due to human error. There was a risk of hitting.

  In the blood cell component separation structure disclosed in Patent Document 2, blood is separated by centrifugal force, and separation and liquid feeding are possible by operating the center of rotation on the machine side, but the apparatus becomes complicated and expensive. There is a problem.

The present invention is as follows.
(1) A microchannel device having a mechanism for filtering a blood sample (A) containing a blood cell component, the opening (B) for introducing the blood sample (A), and the lower part of the opening (B) An inclined membrane filter (C) installed on the side, a support film (D) for holding the inclined membrane filter (C) installed on the lower side of the inclined membrane filter (C), and a support film (D ), A space (E) for storing filtered blood components, a flow path (F) connected to the space (E), and a flow path (F) disposed on the opposite side of the space (E). The inclined membrane filter (C) has a distribution in which the filter diameter gradually decreases from the opening (B) side to the space (E) side, The contact angle with water is 60 degrees or more And in the support film (D) is to have the following holes 500μm or more in diameter 1μm at a ratio of 60% or less aperture ratio of 5% or more, the contact angle with water of the flow path (F) is 60 degrees or less A microchannel device with a blood filtration function, characterized in that
(2) The filtration diameter on the opening (B) side of the inclined membrane filter (C) is 5 μm or more and 20 μm or less, and the filtration diameter on the space (E) side is 0.01 μm or more and 2 μm or less (1) Micro flow channel device with blood filtration function.
(3) The microchannel device with a blood filtration function according to (1) or (2), wherein the thickness of the inclined membrane filter (C) is 50 μm or more and 400 μm or less.
(4) The microchannel device with a blood filtration function according to any one of (1) to ( 3 ), wherein the channel width and the channel depth of the cross-sectional shape of the channel (F) are 1 μm or more and 1 mm or less.
( 5 ) The microchannel device with blood filtration function according to (1), wherein the channel (F) is subjected to at least one treatment selected from plasma treatment, corona discharge treatment, and surface coating treatment with a hydrophilic polymer. .
( 6 ) Surface coating treatment of at least one polymer selected from the group consisting of polyethylene glycol (PEG), eval (EVOH), poval (PVOH), and a polymer having a phosphorylcholine group as a component. ( 1 ) The microchannel device with a blood filtration function according to ( 1 ).
( 7 ) The microchannel with blood filtration function according to any one of (1) to ( 6 ), wherein a site for further reacting and detecting the liquid after separating the blood sample (A) is provided in the flow path (F). Channel device.
( 8 ) A method of using the microfluidic device with blood filtration function according to any one of (1) to ( 7 ), wherein the blood component (A) is introduced into the opening (B) at a pressure of 5.0 kPa to 200 kPa. A method of using a microchannel device that applies a pressure to an opening (B) to filter a blood sample (A) and send the filtered liquid to a space (E).

  In the blood cell component separation structure disclosed in Patent Document 4, a micro slit is provided with a small slit of about 0.1 μm to 2 μm, and a cavity of about 2 μm to 10 μm is provided so that the slit is not clogged with blood cell components. A blood cell component can be filtered from a very small amount of blood sample. However, the process of providing a minute slit with a gap of 0.1 μm to 2 μm in the microchannel is not easy and industrially difficult, and there is a problem that a chip having this structure becomes expensive.

  As described above, a microchannel device with a blood filtration function capable of separating and filtering blood cell components from a blood sample containing a very small amount of blood cell components without clogging of the filter due to the blood cell components by an inexpensive and simple operation, and the blood filtration There was no microchannel device with a function and biosensor chip using the device.

JP 2004-109099 A JP 2006-110491 A US Pat. No. 6,319,719 International Publication WO 2004-097393 Pamphlet

    An object of the present invention is to provide a microchannel device with a blood filtration function capable of separating and filtering blood cell components from a blood sample containing a trace amount of blood components without clogging the filter with blood cell components by an inexpensive and simple operation. That is.

The present invention is as follows.
(1) A microchannel device having a mechanism for filtering a blood sample (A) containing a blood cell component, the opening (B) for introducing the blood sample (A), and the lower part of the opening (B) An inclined membrane filter (C) installed on the side, a support film (D) for holding the inclined membrane filter (C) installed on the lower side of the inclined membrane filter (C), and a support film (D ), A space (E) for storing filtered blood components, a flow path (F) connected to the space (E), and a flow path (F) disposed on the opposite side of the space (E). The inclined membrane filter (C) has a distribution in which the filter diameter gradually decreases from the opening (B) side to the space (E) side, The support film (D) is straight Microchannel device with hemofiltration function characterized by having a 500μm below the through-hole over 1μm in a proportion of less than 60% aperture ratio of 5% or more.
(2) The filtration diameter on the opening (B) side of the inclined membrane filter (C) is 5 μm or more and 20 μm or less, and the filtration diameter on the space (E) side is 0.01 μm or more and 2 μm or less (1) Micro flow channel device with blood filtration function.
(3) The microchannel device with a blood filtration function according to (1) or (2), wherein the thickness of the inclined membrane filter (C) is 50 μm or more and 400 μm or less.
(4) The microchannel device with a blood filtration function according to any one of (1) to (3), wherein a contact angle of the inclined membrane filter (C) with respect to water is 60 degrees or less.
(5) The microchannel device with a blood filtration function according to any one of (1) to (4), wherein the channel width and the channel depth of the cross-sectional shape of the channel (F) are 1 μm or more and 1 mm or less.
(6) The microchannel device with a blood filtration function according to any one of (1) to (5), wherein a contact angle of the channel (F) with respect to water is 60 degrees or less.
(7) The microchannel device with blood filtration function according to (6), wherein the channel (F) is subjected to at least one treatment selected from plasma treatment, corona discharge treatment, and surface coating treatment with a hydrophilic polymer. .
(8) Surface coating treatment of at least one polymer in which the flow path (F) is selected from polyethylene glycol (PEG), eval (EVOH), poval (PVOH), and a hydrophilic polymer whose component is a polymer having a phosphorylcholine group (6) The microchannel device with a blood filtration function according to (6).
(9) The microchannel with blood filtration function according to any one of (1) to (8), wherein the flow path (F) is further provided with a site for reacting and detecting the liquid after separating the blood sample (A). Channel device.
(10) A method of using the microfluidic device with a blood filtration function according to any one of (1) to (9), wherein the blood component (A) is introduced into the opening (B) at a pressure of 5.0 kPa to 200 kPa A method of using a microchannel device that applies a pressure to an opening (B) to filter a blood sample (A) and send the filtered liquid to a space (E).

    According to the present invention, it is possible to obtain a microchannel device capable of separating and filtering blood cell components from a blood sample containing a small amount of blood components without clogging the filter with blood cell components by an inexpensive and simple operation. In addition, the present invention can be applied to various biosensor chips using the microchannel device with blood filtration function.

Hereinafter, an example of an embodiment for carrying out the present invention will be described with reference to the drawings.
FIG. 1 is a schematic plan view of an example of the blood separation chip according to the present invention, and FIG. 2 is a cross-sectional view in the I-II direction of FIG. The microfluidic device shown in FIGS. 1 and 2 has an opening (B) 2 for introducing a blood sample (A) containing a blood cell component formed on a base material 1, and an opening (B) for filtering the blood cell component from the blood sample. From 5 to 60 through holes with a diameter of 1 μm or more and 500 μm or less that support the inclined membrane filter and through which the blood component is filtered. % Of the support film (D) 6, the space (E) 7 in which the blood components are filtered by the membrane filter (C), and the flow path (F) connected to the space (E) 3. It is comprised from the exit (G) 4 arrange | positioned on the opposite side to space (E) and connected with a flow path (F).

  A blood sample (A) containing a blood cell component is a gradient membrane filter (C) 5 in which the filter diameter gradually decreases from the opening (B) 2 through which the blood sample A containing the blood cell component is introduced to the opening (B). Introduced above. After the introduction, with the opening (B) sealed, a constant pressure is applied to the inclined membrane filter (C) into which the blood sample (A) containing blood cell components has been introduced with compressed air or the like. The blood sample (A) containing the blood cell component subjected to pressure is filtered by the inclined membrane filter (C) and passes through. The blood component from which the passed blood cell component has been removed passes through the support film (D) 6 and is stored in a space (E) 7 for storing the blood component from which the blood cell component has been filtered by the membrane filter (C). Thereafter, the blood component obtained by filtering the blood cell component stored in the space (E) flows into the flow path (F) 3 by affinity with the flow path surface, capillary effect, or external power by a pump or the like, and the space (E ) And the blood sample (A) containing blood cell components can be filtered by reaching the outlet (G) 4 disposed on the opposite side to the flow path (F).

    FIG. 3 is a schematic plan view of an example of the blood separation chip of the present invention, and FIG. 4 is a cross-sectional view taken along the line II-II of FIG. The microfluidic device shown in FIGS. 3 and 4 has an opening (B) 2 for introducing a blood sample (A) containing a blood cell component formed in the base material 1, and an opening (B) for filtering the blood cell component from the blood sample. From 5 to 60 through holes with a diameter of 1 μm or more and 500 μm or less that support the inclined membrane filter and through which the blood component is filtered. % Of the support film (D) 6 having a ratio of not more than%, a flow path connected to the space (E) 7 and the space (E) from the outer periphery for collecting blood components filtered by the membrane filter (C) ( F) 3, an outlet (G) 4 disposed on the opposite side of the space (E) and communicated with the flow path (F), provided to facilitate the flow of blood components filtered from the space E into the flow path (F) Air vent flow path 8, And a air vent holes 9 to contact A vent channel and the outside.

  A blood sample (A) containing a blood cell component is a gradient membrane filter (C) 5 in which the filter diameter gradually decreases from the opening (B) 2 through which the blood sample A containing the blood cell component is introduced to the opening (B). Introduced above. After the introduction, with the opening (B) sealed, a constant pressure is applied to the inclined membrane filter into which the blood sample (A) containing blood cell components has been introduced with compressed air or the like. The blood sample (A) containing a blood cell component subjected to pressure is filtered through the blood cell component with an inclined membrane filter. The blood component from which the passed blood cell component has been removed passes through the support film (D) 6 and is stored in a space (E) 7 for storing the blood component from which the blood cell component has been filtered by the membrane filter (C). Thereafter, the blood component obtained by filtering the blood cell component stored in the space (E) flows into the flow path (F) 3 by affinity with the flow path surface, capillary effect, or external power by a pump or the like, and the space (E In order to facilitate the flow of the blood component filtered from the space (E) to the flow path (F), which reaches the outlet (G) 4 disposed on the opposite side to the flow path (F). By providing the air vent channel 8 provided and the air vent hole 9 connecting the air vent channel and the outside, the blood component smoothly filtered flows into the channel (F), and a blood sample containing a blood cell component (A) can be filtered.

FIG. 5 is a schematic plan view of an example of the blood separation chip of the present invention, and FIG. 6 is a cross-sectional view taken along the line II-II of FIG. The microfluidic device shown in FIGS. 5 and 6 has an opening (B) 2 for introducing a blood sample (A) containing a blood cell component formed in the base material 1, and an opening (B) for filtering the blood cell component from the blood sample. From 5 to 60 through holes with a diameter of 1 μm or more and 500 μm or less that support the inclined membrane filter and through which the blood component is filtered. % Of the support film (D) 6 having a ratio of not more than%, a flow path connected to the space (E) 7 and the space (E) from the outer periphery for collecting blood components filtered by the membrane filter (C) ( F) 3, a reaction chamber and a detection unit 10 arranged on the opposite side of the space (E) and communicated with the flow channel (F), a flow channel 31 connected to the reaction chamber and the detection unit 10, a reaction chamber and An outlet (G) 4 installed on the opposite side of the detection unit 10 and communicating with the flow path 31 ; an air vent flow path 8 provided to facilitate the flow of blood components filtered from the space E to the flow path (F); It consists of an air vent hole 9 that communicates the air vent channel with the outside.

A blood sample (A) containing a blood cell component is a gradient membrane filter (C) 5 in which the filter diameter gradually decreases from the opening (B) 2 through which the blood sample A containing the blood cell component is introduced to the opening (B). Introduced above. After the introduction, with the opening (B) sealed, a constant pressure is applied to the inclined membrane filter into which the blood sample (A) containing blood cell components has been introduced with compressed air or the like. The blood sample (A) containing a blood cell component subjected to pressure is filtered through the blood cell component with an inclined membrane filter. The blood component from which the passed blood cell component has been removed passes through the support film (D) 6 and is stored in a space (E) 7 for storing the blood component from which the blood cell component has been filtered by the membrane filter (C). Thereafter, the blood component in which the blood cell component stored in the space (E) is filtered flows into the flow path (F) 3 by affinity with the flow path surface, capillary effect, or external power by a pump or the like. E) the flow into the reaction chamber and the detection unit 10 is contacted with the opposite disposed side channel (F), the reaction chamber and the detection unit 10 is the flow path 31 connected with, on the opposite side of the reaction chamber and the detection unit 10 It flows to the outlet (G) 4 that is installed and communicates with the flow path 31 . At this time, an air vent channel 8 provided for facilitating the flow of blood components filtered from the space (E) to the channel (F), and an air vent hole 9 connecting the air vent channel and the outside are provided. Thus, the smoothly filtered blood component flows into the flow path (F), and the blood sample (A) containing the blood cell component can be filtered.
In the reaction chamber and the detection unit 10, the blood cell component that has flowed in by immobilizing a reagent that reacts with the filtered blood component, an antibody against the detected protein, etc. is reacted with the filtered blood component. The resulting output can be detected. The output obtained by the reaction is absorbance in the case of a color reaction, fluorescence in the case of using a fluorescent label, current in the case of an electrochemical reaction, and a generally used method can be used.

  The microchannel device of the present invention preferably processes a flat base material. Examples of the material of the base material include Si, glass, and plastic. Of these, plastic is particularly preferable. As the plastic material, including the processability and economy, the most used detection method is fluorescence detection, so there is little autofluorescence, and the bioadaptive material from FDA (no problem even if it comes into contact with blood). Cyclic polyolefin resin (COC) that is recognized as the most preferred, but it is possible to select various plastic materials, use of the device to be produced, treatment, solution used, physiologically active substance, characteristics of the detection method In consideration of moldability, heat resistance, chemical resistance, adsorptivity and the like, it is appropriately selected.

    Examples of plastic materials include polystyrene, polyethylene, polyvinyl chloride, polypropylene, polycarbonate, polyester, polymethyl methacrylate, polyvinyl acetate, vinyl-acetate copolymer, styrene-methyl methacrylate copolymer, and acrylonitrile-styrene copolymer. Examples include coalesce, acrylonitrile-butadiene-styrene copolymer, nylon, polymethylpentene, silicone resin, amino resin, polysulfone, polyethersulfone, polyetherimide, fluororesin, polyimide, and the like. In addition, additives such as pigments, dyes, antioxidants, and flame retardants may be appropriately mixed with these plastic materials.

    In the present invention, the processing method for processing the inlet, the flow path, and the like into the base material is arbitrary, and methods such as cutting, injection molding, solvent casting, photolithography, laser ablation, and hot embossing can be used.

  The shape, size, and depth of the opening (B) for introducing the blood sample (A) are the amount of the blood sample (A) containing the blood cell component to be filtered, the thickness of the filter, and the inclined membrane filter (C). Is appropriately determined depending on the filtration diameter.

  The membrane filter (C) should be an inclined type in which the filtration diameter of the filter gradually decreases from the opening (B) side to the space (E) side. A uniform membrane filter that does not gradually reduce the filtration diameter is filtered from blood samples. When the filter is used, clogging occurs, or a high pressure is required to pass the blood component through the filter, resulting in a problem (hemolysis) that destroys the blood cell component by the pressure for filtering the blood cell component.

  The filtration diameter on the opening side of the inclined membrane filter (C) is preferably 5 μm or more and 20 μm or less, more preferably 8 μm or more and 15 μm or less. If the filtration diameter on the opening side of the inclined membrane filter (C) is less than the lower limit value, the blood cell component is clogged with the filter, and the pressure required for filtering the blood sample increases or the blood cell component is increased by increasing the pressure. The problem of destruction (hemolysis) occurs. If the upper limit is exceeded, the function of sufficiently filtering blood cell components contained in the blood sample will not be exhibited.

  The filtration diameter on the space (E) side of the inclined membrane filter (C) is preferably 0.01 μm or more and 2 μm or less, and more preferably 0.05 μm or more and 1.5 μm or less. When the filtration diameter on the space (E) side of the inclined membrane filter (C) is less than the lower limit value, blood cell components are clogged with the filter, and the pressure required for filtering the blood sample increases, or by increasing the pressure The problem of destruction of blood cell components (hemolysis) occurs. When the filtration diameter exceeds the upper limit, the function of sufficiently filtering blood cell components contained in the blood sample is not exhibited.

  The thickness of the inclined membrane filter (C) is preferably 50 μm or more and 400 μm or less, more preferably 100 μm or more and 300 μm or less. If the thickness of the inclined membrane filter is less than the lower limit value, the filtration accuracy for filtering the blood sample is deteriorated, and it is difficult to adjust the pressure required for filtration without breaking the blood cell component while maintaining the filtration accuracy. If the thickness of the inclined membrane filter exceeds the upper limit value, many blood components remain in the inclined membrane filter (C), making it difficult to efficiently collect blood components.

  The contact angle of the inclined membrane filter (C) with respect to water is preferably 60 degrees or less, and more preferably 40 degrees or less. If the contact angle of the inclined membrane filter (C) with water exceeds the upper limit, the blood sample will not flow easily through the fine filtration diameter, and the pressure required for filtration will increase and it will take time to filter. Is likely to occur.

  The support film (D) for holding the filter needs to have through holes having a diameter of 1 μm or more and 500 μm or less in a ratio of an opening ratio of 5% or more and 60% or less, and preferably a through hole having a diameter of 10 μm or more and 300 μm or less. At an opening ratio of 10% to 50%.

  When the support film (D) having a diameter of the through-hole of less than the lower limit value or an opening rate of less than the lower limit value is used, the blood component filtered by the inclined membrane filter (C) is high to flow to the opening (E). Pressure is required, and when a high pressure is used, the blood sample (A) is filtered through the inclined membrane filter (C), causing a problem that the blood cell component is destroyed and functions as a real blood separation chip. No longer.

  When the support film (D) having a diameter of the through-hole exceeding the upper limit value or the opening ratio exceeding the upper limit value is used, the support film (D) cannot support the inclined membrane filter (C), and the blood sample (A ), When the pressure is applied to the inclined membrane filter (C), the inclined membrane filter (C) is broken and the function of filtering blood cell components cannot be exhibited.

  The constant pressure applied after introducing the blood component (A) into the opening (B) is preferably 5.0 kPa or more and 200 kPa or less, more preferably 10 kPa or more and 150 kPa or less. When pressure below the lower limit is applied, the blood component is difficult to be filtered and flow through the inclined membrane filter and the support film (D) for holding the filter. When pressure exceeding the upper limit is applied, blood cell components are destroyed and blood This is not good because it causes a problem that the sample cannot be separated into blood cell components and blood components.

  The channel width and the channel depth of the cross-sectional shape of the channel (F) are preferably 1 μm or more and 1 mm or less, more preferably 5 μm or more and 800 μm or less. If the flow path width and flow path depth of the flow path (F) are less than the lower limit values, a technique is required for producing the flow path, and it cannot be industrially produced and is not efficient. If the flow path width and the flow path depth of the flow path (F) exceed the upper limit values, the fluid movement effect due to the surface tension and the capillary effect becomes thin, and it takes time for the liquid to flow through the flow path.

  It is preferable that the contact angle with respect to the water of a flow path (F) is 60 degrees or less, More preferably, it is 40 degrees or less. If the contact angle of the flow path (F) with respect to water exceeds the upper limit value, the fluid movement effect due to the surface tension or the capillary effect becomes thin, and it takes time for the liquid to flow through the flow path.

  As a method of setting the contact angle with respect to the flow path (F) to 60 degrees or less, plasma treatment or corona discharge treatment, hydrophilic polymer surface coating treatment, polyethylene glycol (PEG), Eval (EVOH), poval (PVOH) ), It is preferable to surface coat a hydrophilic polymer containing a polymer having a phosphorylcholine group as a component, but there is no particular limitation as long as the method does not adversely affect the composition or composition ratio of blood components.

  The flow path (F) can be provided with a space in which reaction reagents and electrodes that react with the blood components after the blood sample (A) is separated are provided to analyze the blood components.

    The flow path design of these blood separation chips is appropriately designed in consideration of the detection object and convenience. Equipped with membranes, pumps, valves, sensors, motors, mixers, gears, clutches, microlenses, electrical circuits, etc. as blood separation chips, or composites by processing multiple microchannels on the same substrate It is also possible.

EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to these examples.
Example 1
A slide flat plate (1 mm thick × 30 mm × 70 mm) made of cyclic polyolefin resin (COC) is used as a base material, and the cutting method is performed in the shape shown in FIG. The upper member 11 was produced by carrying out processing using Similarly, the shape shown in FIG. 10, which is a cross-sectional view taken along the line I-II in FIGS. 9 and 9, was processed using a cutting method to produce an intermediate member 12. The diameter of the opening (B) 2 for introducing the blood sample (A) containing blood cell components and the space (E) 7 for storing the filtered liquid is processed as a through hole of 4 mmφ, and the diameter of the outlet (G) 4 is It was processed as a 1 mmφ through hole. The flow path (F) 3 connecting the space (E) for storing the filtered liquid and the outlet (G) has a rectangular cross section, a flow path width of 0.5 mm, a flow path depth of 0.1 mm, and a flow path length of 20 mm. It was. Hydrophilic treatment by oxygen plasma treatment was performed on the flow path of the processed COC resin substrate, and the contact angle between the obtained COC substrate and water was 40 degrees. The inclined membrane filter (C) 5 is a nylon membrane filter with a thickness of 200 μm and a diameter of 5 mmφ, with a filter diameter of 10 μm on the opening (B) side and a filter diameter of 0.2 μm on the space (E) side. It was used. The contact angle between this filter and water was measured and found to be 20 degrees. As the support film (D) 6 for holding the filter, a fluorine film having a thickness of 100 μm and a diameter of 5 mmφ in which through-holes having a diameter of 100 μm were opened at an opening ratio of 20% was used.

  These members are arranged as shown in FIG. 11, and at the bottom are slide flat plates (0.7 mm thickness × 30 mm × 70 mm) made of cyclic polyolefin resin (COC) that has not been processed such as flow paths. The lower member 13 was placed, and the substrate was attached by thermocompression bonding for 20 minutes while applying a load of 200 kg with a hot plate at 100 ° C. to produce a microchannel device 14 shown in FIG.

  5 μL of blood was introduced into the opening (B) of the microchannel device 14, and after introduction, a pressure of 20 kPa was applied with air in the shape shown in FIG. 13 that sealed the opening (B). A few seconds after the pressure was applied, the blood component from which the blood cell component had been filtered flowed into the flow path.

(Example 2)
A slide-like flat plate (1 mm thick × 30 mm × 70 mm) made of polystyrene resin (PS) is used as a base material, and a cutting method is used for the shape shown in FIG. 8 which is a cross-sectional view in the I-II direction of FIGS. Then, the upper member 11 was manufactured by processing. Similarly, the shape shown in FIG. 10, which is a cross-sectional view taken along the line I-II in FIGS. 9 and 9, was processed using a cutting method to produce an intermediate member 12. The diameter of the opening (B) 2 for introducing the blood sample (A) containing blood cell components and the space (E) 7 for storing the filtered liquid is processed as a through hole of 4 mmφ, and the diameter of the outlet (G) 4 is It was processed as a 1 mmφ through hole. The flow path (F) 3 connecting the space (E) for storing the filtered liquid and the outlet (G) has a rectangular cross section, a flow path width of 0.3 mm, a flow path depth of 0.1 mm, and a flow path length of 20 mm. It was. Hydrophilic treatment was performed by coating MPC polymer (manufactured by NOF Corporation) on the flow path of the processed PS resin substrate. The contact angle between the obtained PS substrate and water was 50 degrees. The inclined membrane filter (C) 5 is a nylon membrane filter having a thickness of 300 μm and a diameter of 5 mmφ. The filter diameter on the opening (B) side is 8 μm, and the filter diameter on the space (E) side is 0.05 μm. I used something. The contact angle between this filter and water was measured and found to be 20 degrees. As a support film (D) 6 for holding the filter, a polyethylene terephthalate having a diameter of 50 μm and a diameter of 5 mmφ in which about 50 μm holes knitted with polyethylene terephthalate (PET) having a diameter of 50 μm are opened at an opening ratio of 25%. PET) mesh was used.

  These members are arranged as shown in FIG. 11, and a lower member 13, which is a slide-like flat plate (0.7 mm thickness × 30 mm × 70 mm) made of PS that is not subjected to processing such as a flow path, is provided at the bottom. The microchannel device 14 shown in FIG. 12 was manufactured by placing and bonding the substrate by thermocompression bonding for 20 minutes while applying a load of 150 kg with a 95 ° C. hot plate.

  5 μL of blood was introduced into the opening (B) of the microchannel device 14, and after introduction, a pressure of 30 kPa was applied with air in the shape shown in FIG. 13 that hermetically seals the opening (B). A few seconds after the pressure was applied, the blood component from which the blood cell component had been filtered flowed into the flow path.

(Comparative Example 1)
Compared to Example 1, a microchannel device was prepared in the same manner as in Example 1 except that a 100 μm thick nylon membrane filter having a uniform distribution with a filter filtration diameter of 3 μm was used instead of the inclined filter (C). Was made.
Although 5 μL of blood was introduced into the opening (B) of the microchannel device and the opening (B) was sealed after introduction, a pressure of 1 to 100 kPa was applied with air in the shape shown in FIG. The filtered blood component did not flow into the flow path. Further, when a pressure of 100 kPa or more was applied with air, the blood cell component was destroyed and the red colored blood component flowed into the flow path, and blood separation was not successful.

(Comparative Example 2)
Compared to Example 1, a microchannel device was prepared in the same manner as in Example 1 except that a 150 μm thick nylon membrane filter having a uniform distribution with a filter filtration diameter of 6 μm was used instead of the inclined filter (C). Was made.
In the shape shown in FIG. 13, 5 μL of blood is introduced into the opening (B) for introducing the blood sample (A) containing the blood cell component of the microchannel device, and the opening (B) is sealed after the introduction. When a pressure of 20 kPa was applied with air, a red colored blood component flowed into the flow path. As a result of examining the blood component flowing out into the flow path with a microscope, it was found that a large amount of red blood cells were contained and the red blood cell components could not be separated well.

(Comparative Example 3)
Compared with Example 1, a microchannel device was produced in the same manner as in Example 1 except that the support film (D) holding the filter was not used.
When 5 μL of blood is introduced into the opening (B) of the microchannel device and the opening (B) is sealed after introduction, a pressure of 20 kPa is applied with air in the shape shown in FIG. The torn blood separation chip did not function.

(Comparative Example 4)
Compared to Example 1, as the support film (D) for holding the filter, a polyethylene terephthalate (PET) film having a thickness of 50 μm and a diameter of 5 mmφ in which through holes having a diameter of 10 μm are opened with an aperture ratio of 3% is used. Produced a microchannel device in the same manner as in Example 1.
Blood 5 μL was introduced into the opening (B) of the above microchannel device, and after the introduction, the opening (B) was sealed. In the shape shown in FIG. The blood component did not come out to the opening (E) where the water was accumulated, and did not function as a blood separation chip.

    By using the present invention, a microchannel device with a blood filtration function capable of separating and filtering blood cell components from a blood sample containing a small amount of blood cell components without clogging of the filter due to blood cell components by an inexpensive and simple operation, and this Biosensor chip using micro-channel device with blood filtration function can be provided, and clinical tests in the fields of chemistry, biochemistry, medicine, etc., especially point-of-care testing, or blood separation for home use etc. is required It can be used for biochips used for measurement.

Outline plan view of an example of the microchannel device of the present invention I-II sectional view of FIG. Outline plan view of an example of the microchannel device of the present invention I-II direction sectional view of FIG. Outline plan view of an example of the microchannel device of the present invention I-II sectional view of FIG. Processing plan view of the upper member of the microchannel device used in the examples I-II direction sectional view of FIG. Processing plan view of the middle member of the microchannel device used in the examples I-II sectional view of FIG. Example of bonding layout of microchannel device used in examples Sectional drawing of the microchannel device after bonding of FIG. Sectional drawing which shows an example of the usage method of the microchannel device of this invention

Explanation of symbols

1 Base material of microchannel device 2 Opening (B) for introducing blood sample (A) containing blood cell components
3 Flow path (F) communicating from the opening (B)
4 Exit (G)
5 Inclined membrane filter (C)
6 Support film for holding the filter (D)
7 Opening for storing filtered blood components (E)
8 Air vent channel connecting space (E) and air vent hole 9 Air vent channel connecting air vent channel and outside 10 Chamber 11 in which filtered blood component reacts and is detected Upper member 12 Middle member 13 Lower Member 14 Micro-channel device 21 Pipe 22 for sending air to apply pressure Jig 23 for applying pressure to filter blood sample (A) Packing 24 for maintaining airtightness Blood sample (A)
31 Flow path connected to reaction chamber and detector

Claims (8)

A microchannel device having a mechanism for filtering a blood sample (A) containing a blood cell component, which is installed on the lower side of the opening (B) and the opening (B) for introducing the blood sample (A) Tilted membrane filter (C), support film (D) installed on the lower side of the tilted membrane filter (C) to hold the tilted membrane filter (C), lower part of the support film (D) A space (E) for storing filtered blood components installed on the side, a flow path (F) connected to the space (E), and a flow path (F) disposed on the opposite side of the space (E) The inclined membrane filter (C) has a distribution in which the filter diameter gradually decreases from the opening (B) side to the space (E) side, and is in contact with water. The angle is less than 60 degrees , The support film (D) is to have the following holes 500μm or more in diameter 1μm at a ratio of 60% or less aperture ratio of 5% or more, the contact angle with water of the flow path (F) is less than 60 degrees A microchannel device with a blood filtration function. The blood filtration according to claim 1, wherein the filtration diameter on the opening (B) side of the inclined membrane filter (C) is 5 µm or more and 20 µm or less, and the filtration diameter on the space (E) side is 0.01 µm or more and 2 µm or less. Functional microchannel device. The microchannel device with a blood filtration function according to claim 1 or 2, wherein the inclined membrane filter (C) has a thickness of 50 µm or more and 400 µm or less. The microchannel device with a blood filtration function according to any one of claims 1 to 3 , wherein the channel width and channel depth of the cross-sectional shape of the channel (F) are 1 µm or more and 1 mm or less. The microchannel device with a blood filtration function according to claim 1, wherein the channel (F) is subjected to at least one treatment selected from plasma treatment, corona discharge treatment, and surface coating treatment with a hydrophilic polymer. The flow path (F) is surface-coated with at least one polymer selected from polyethylene glycol (PEG), eval (EVOH), poval (PVOH), and a hydrophilic polymer whose component is a polymer having a phosphorylcholine group. The microchannel device with a blood filtration function according to claim 5 . The microchannel device with blood filtration function according to any one of claims 1 to 6, wherein a site for reacting and detecting the liquid after separating the blood sample (A) is further provided in the channel (F). A claim 1-7 Using the blood filtration function microfluidic device according to any one, opening the 200kPa pressure below than 5.0kPa after introduction into the blood component (A) opening (B) ( A method of using a micro-channel device for filtering a blood sample (A) by applying to B) and sending the filtered liquid to space (E).

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