JP2007071655A - Cartridge - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cartridge suitable for use as the so-called a disposable type and enabling easy and accurate dilution. <P>SOLUTION: The cartridge A is equipped with: a liquid introducing port 3 into which a sample liquid is introduced; and a dilution means 4 which includes a dilution liquid tank 41 storing a diluting liquid 40 for diluting the sample liquid, a sample liquid metering means 43 for separating a definite amount of the sample liquid introduced from the liquid introducing port 3, and a dilution tank 42A for mixing the sample liquid with at least a part of the dilution liquid, and the cartridge is mounted on a separate apparatus. A sample liquid metering means 4 includes: the introducing flow channel 43a extended from a liquid introducing port 3; the metering flow channel 43c connected to the introducing flow channel 43a through a branch part 43b; and an overflow flow channel 43d, and a metering flow channel 43c goes toward the dilution tank 42A. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a cartridge such as a cartridge for an analyzer loaded in an analyzer for analyzing a specific component in blood, for example, and particularly relates to a disposable type cartridge.

  Analyzing specific components in the blood is effective in understanding the health condition of the human body or treating specific diseases. A blood analyzer used for such an application includes a so-called blood cell counter that counts blood cells such as red blood cells or white blood cells in blood.

  FIG. 26 shows an example of a cartridge loaded in a conventional blood cell counter. The cartridge X shown in the figure includes a main body 91, a diluent tank 92, an introduction part 93, a dilution tank 94, a recovery tank 95, a measuring tank 96, and a suction port 98. When blood 99 is introduced into the introduction port 93a, blood 99 permeates into the flow paths 93c and 93d by capillary action. When the rotating member 93b is rotated 90 ° in this state, the portion of the blood 99 contained in the flow path 93d is separated. Next, when suction is performed from the suction port 98, the diluent 92 a in the dilution tank 92 and the blood 99 in the flow path 93 d are sent to the dilution tank 94. In the process of being sent to the dilution tank 94, the diluent 92a and the blood 99 are mixed. As a result, diluted specimen blood is generated in the dilution tank 94. A partition wall 97 having pores is provided between the dilution tank 94 and the recovery tank 95. When the suction from the suction port 98 is continued, the sample blood in the dilution tank 94 flows into the recovery tank 95 through the pores. The dilution tank 94 and the collection tank 95 are provided with electrodes 94a and 95a. The resistance between the electrodes 94a and 95a is monitored while the sample blood passes through the pores. For example, since red blood cells are insulators, the resistance decreases each time the red blood cells pass through the pores. By counting the number of drops, the number of red blood cells contained in the sample blood can be counted. The specimen blood that has flowed into the collection tank 95 is further sent to the measurement tank 96. Before and after the flow direction of the measuring tank 96, flow rate detection means (not shown) by electrical or optical means is provided. By this flow rate detection means, the measured amount of the sample blood is obtained. With the above operation, the number of red blood cells in blood 99 is counted. The cartridge X has a relatively simple structure, and is configured as a so-called disposable type cartridge that is used only once.

  However, in order for a user to frequently count blood cells using a blood cell counter using the cartridge X, there are the following problems.

  First, in the introduction part 93, the blood 99 is measured by rotating the rotary member 93b. The rotary member 93b is required to be a sealable member that is separate from the main body 91 but does not have a gap between the main body 91 and the like. This is because the leakage of blood 99 not only lowers the test accuracy but also may deteriorate the sanitary condition. It is difficult to manufacture such a rotating member 93b, which complicates the manufacturing process of the cartridge X. Further, it is difficult to rotate the rotating member 93b by a driving source outside the cartridge X without liquid leakage.

  Moreover, the diluent 92a and the blood 99 may not be sufficiently mixed only by pouring the diluent 92a and the blood 99 together into the dilution tank 94. For example, if the blood cell component is biased to the corner of the dilution tank 94, the blood cell component is not properly sent out to the collection tank 95. In such a case, the counting accuracy of red blood cells is lowered.

International pamphlet W / O 03/104771

  The present invention has been conceived under the circumstances described above, and is suitable for use as a so-called disposable type, and its object is to provide a cartridge that can be easily and accurately diluted. To do.

  In order to solve the above problems, the present invention takes the following technical means.

  The cartridge provided by the present invention includes a liquid introduction port into which a sample liquid is introduced, a diluent tank in which a diluent for diluting the sample liquid is stored, and the sample liquid introduced from the liquid introduction port. A diluting means including a sample liquid measuring means for separating a predetermined amount from the sample liquid and one or more dilution tanks for mixing at least part of the sample liquid and the diluting liquid. The sample liquid metering means includes an introduction channel extending from the liquid introduction port, a metering channel and an overflow channel connected to the introduction channel via a branch portion. The metering channel is directed to the dilution tank.

  According to such a configuration, when the measurement is performed using the dilution liquid measuring means, for example, it is not necessary to rotate the rotating member having a channel having a predetermined length. Therefore, there is no risk of liquid leakage in the weighing operation. Further, the cartridge can be easily manufactured.

  In a preferred embodiment of the present invention, a stirring bar is accommodated in the dilution tank. According to such a configuration, for example, the sample liquid and the diluted liquid can be appropriately mixed in the dilution tank by rotating the stirring bar using a magnetic force generation source.

  In a preferred embodiment of the present invention, an orifice is interposed between the metering channel and the dilution tank. Such a configuration is advantageous for retaining a predetermined amount of the sample solution in the measuring flow channel.

  In a preferred embodiment of the present invention, at least a portion corresponding to the length of the metering channel from the branch portion in the overflow channel is equal to or larger than the cross-sectional area of the metering channel. Is also considered small. According to such a configuration, it is possible to prevent most of the sample liquid from flowing into the overflow channel.

  In a preferred embodiment of the present invention, the diluting means includes diluting liquid measuring means for separating a predetermined amount from the diluting liquid in the diluting liquid tank. According to such a configuration, for example, it is suitable for use in applications where it is necessary to dilute at an accurate dilution rate, such as counting blood cells in blood or pretreatment thereof.

  In a preferred embodiment of the present invention, the diluent metering means includes a metering channel having a large cross section and a pair of tapered portions connected to both ends of the large cross section in the flow direction. According to such a configuration, it is possible to retain the dilution liquid in a relatively large amount in the measurement channel. Thereby, for example, a high magnification dilution of about 100 times can be performed. Moreover, when the dilution liquid flows into the large cross-section portion or flows out from the large cross-section portion, it is possible to prevent the flow from being unduly disturbed.

  In a preferred embodiment of the present invention, the dilution means includes first and second dilution tanks, and a flow into which the sample liquid introduced from the liquid introduction port flows into the first dilution tank. And a flow path through which the diluent flows from the dilution tank, the second dilution tank has a flow path through which the diluted sample liquid diluted in the first dilution tank flows, A flow path through which the diluent flows from the diluent tank is connected. According to such a configuration, it is possible to dilute the sample solution in so-called two stages. Therefore, for example, by performing 100-fold dilution twice, it is possible to achieve a very high dilution of 10,000-fold dilution.

  In preferable embodiment of this invention, the buffer tank arrange | positioned downstream with respect to the said 1st dilution tank is further provided, and the dry hemolytic agent is apply | coated to the said buffer tank. According to such a configuration, for example, when white blood cells are counted, the influence of red blood cells can be appropriately excluded. Further, the dry hemolyzing agent hardly contains moisture. Therefore, it is possible to prevent problems such as liquid leakage from the cartridge. It is also preferable for reducing the weight of the cartridge.

  In a preferred embodiment of the present invention, by providing one or more analysis units for analyzing a specific component contained in a diluted sample solution diluted by the dilution means, the specific component contained in the sample solution is provided. It is configured as a cartridge for an analyzer loaded in an analyzer for performing analysis. According to such a configuration, the preparation of the diluted sample solution and the analysis of the specific component can be performed collectively. Therefore, it is suitable for the user to easily perform the above analysis.

  In a preferred embodiment of the present invention, the apparatus further comprises one or more storage means for storing the analyzed diluted sample solution. According to such a configuration, the sample solution can be accurately and easily measured by retaining a predetermined amount of the sample solution in the measurement channel.

  In a preferred embodiment of the present invention, a first analysis unit for analyzing a diluted sample solution diluted in the first dilution tank and a diluted sample solution diluted in the second dilution tank are analyzed. A second analysis unit. According to such a configuration, it is possible to analyze two types of diluted sample solutions having different dilution ratios. As a result, a wide variety of analyzes can be performed on the sample solution.

  In a preferred embodiment of the present invention, a flow rate measurement unit for measuring the flow rate of the diluted sample solution that has passed through the analysis unit is further provided. According to such a configuration, it is possible to accurately grasp the flow rate of the diluted sample solution that has passed through the analysis unit within a predetermined time without flowing the diluted sample solution at a constant flow rate. This is advantageous for analyzing the concentration or number of distributions per volume, and can simplify the pump and the like provided in the analyzer.

  In a preferred embodiment of the present invention, the flow rate measuring unit includes a meandering flow path and two or more diluted sample liquid detection means arranged at positions separated in the flow direction of the meandering flow path. According to such a configuration, it is possible to make the planar arrangement close to a square shape, for example, while making the flow direction length of the flow rate measuring unit relatively long. Therefore, it is possible to improve the analysis accuracy and reduce the size of the cartridge.

  In a preferred embodiment of the present invention, the diluted sample liquid detection means includes an electrode. According to such a configuration, the diluted sample solution can be easily detected.

  In a preferred embodiment of the present invention, the meandering channel serves as the storage means. Such a configuration is advantageous in reducing the size of the cartridge.

  In a preferred embodiment of the present invention, the analysis unit includes an electrical resistance analysis unit having a pore and a pair of electrodes spaced apart from each other. According to such a configuration, for example, it is possible to appropriately count blood cells using fluctuations in electrical resistance.

  In a preferred embodiment of the present invention, the analysis section further includes an optical analysis section having a reflection film, a light transmission section, and a reagent applied to the reflection film or the light projection section. According to such a configuration, analysis of, for example, Hb and CRP can be performed appropriately.

  In a preferred embodiment of the present invention, the channel through which the sample solution and the diluted sample solution are flowed is constituted by a hydrophobic surface having a water contact angle of 60 degrees or more. According to such a configuration, it is possible to prevent the sample solution or the diluted sample solution from being unduly flowing out due to capillary action.

  In a preferred embodiment of the present invention, the flow path includes one having a width / depth of 1 or more and 5 or less. According to such a configuration, the sample solution or the diluted sample solution can be flowed as a uniform flow without unduly meandering in the flow path.

  In a preferred embodiment of the present invention, a main body and a printed wiring board attached to the main body are provided, and the main body has a plurality of recesses or grooves, and the plurality of recesses or grooves A plurality of flow paths or tanks are configured by covering the groove with the printed wiring board. According to such a configuration, the cartridge can have a simple structure and a configuration suitable for performing a wide variety of analyses.

  In a preferred embodiment of the present invention, a body having a plurality of recesses or grooves, an electrode formed integrally with the body by insert molding so as to be exposed in the plurality of recesses or grooves, and the body A plurality of flow paths or tanks are configured by covering the plurality of recesses or grooves with the covering member. According to such a structure, the said main body and the said electrode can be formed collectively, and it is suitable for the improvement of manufacturing efficiency.

  In a preferred embodiment of the present invention, the sample solution is blood.

  In a preferred embodiment of the present invention, the specific component is blood cells such as red blood cells, white blood cells, and platelets.

  In a preferred embodiment of the present invention, the specific component is hemoglobin or C-reactive protein.

  Other features and advantages of the present invention will become more apparent from the detailed description given below with reference to the accompanying drawings.

  Hereinafter, preferred embodiments of the present invention will be specifically described with reference to the drawings.

  1 and 2 show an example of a cartridge according to the present invention. The cartridge A shown in FIG. 1 has a main body 1 and a printed wiring board 2 attached to each other, and includes a liquid inlet 3, a diluting means 4, a plurality of analyzers 5A, 5B, 5C, 5D and two flow rates. It comprises measuring units 6A and 6B, and is configured as a disposable analysis device cartridge to be loaded into an analysis device (not shown).

  The main body 1 has a flat rectangular shape and is made of a transparent resin such as acrylic. On the lower surface of the main body 1 in FIG. 2, a plurality of recesses or grooves for forming channels and tanks to be described later are formed. In the present embodiment, the main body 1 has a size of about 70 mm square and a thickness of about 3 mm.

  In the printed wiring board 2, a plurality of base materials made of epoxy resin or the like are laminated, and a wiring pattern made of copper foil or the like is formed between these base materials. In addition, a plurality of electrodes 51 and 62 described later are formed on the printed wiring board 2. These electrodes 51 and 62 have a so-called through-hole structure. A connector 8 is formed on the extended portion of the printed wiring board 2. The connector 8 is used to connect the cartridge A to an analyzer such as a blood cell counter (not shown). The main body 1 and the printed wiring board 2 are liquid-tightly bonded using, for example, an adhesive. Moreover, as for the main body 1 and the printed wiring board 2, the surface which forms the flow path etc. which are mentioned later at least is made into the hydrophobic surface whose water contact angle is 60 degree | times or more.

  The liquid inlet 3 is for introducing blood to be analyzed into the cartridge A. The liquid introduction port 3 is a through-hole formed in the main body 1 and has a diameter of about 3 mm.

  The diluting means 4 is for diluting the blood introduced from the liquid introducing port 3 to a concentration suitable for various analyses, and includes a diluting liquid tank 41, first and second diluting tanks 42A and 42B, and blood measuring means 43. , And a diluent metering means 44. The diluting means 4 of the present embodiment is of a type capable of two-stage dilution using the first and second dilution tanks 42A and 42B as will be described later.

  The diluent tank 41 is for storing a diluent 40 for diluting blood in the cartridge A. The diluent tank 41 has a diameter of about 12 mm and a depth of about 2 mm, and can contain a diluent 40 of about 200 μL. Diluent 40 is, for example, physiological saline. For example, an aluminum pack having a shape along the inner surface of the diluent tank 41 is used to incorporate the diluent 40 in the diluent tank 41.

  The blood metering means 43 is disposed between the liquid introduction port 3 and the first dilution tank 42A, and includes an introduction channel 43a, a metering channel 43c, and an overflow channel 43d. The introduction flow path 43a is a flow path for introducing blood from the liquid introduction port 3 and has a width of about 250 μm, a depth of about 250 μm, and a width / depth of 1. Each flow path described below has the same width and depth as the introduction flow path 43a, unless otherwise specified. In order to ensure a uniform flow in each flow path, the width / depth is preferably 5 or less. A metering flow path 43c and an overflow flow path 43d extend from the introduction flow path 43a via a branching portion 43b. The measuring channel 43c is for temporarily retaining a predetermined amount of blood suitable for analysis. The measuring channel 43c has a length of about 8 mm and a volume of about 0.5 μL. An orifice 43e is provided between the measurement channel 43c and the first dilution tank 42A. The orifice 43e has a width of about 50 μm, and is intended to intentionally increase the pressure loss resistance from the measuring flow path 43c to the first dilution tank 42A. The overflow channel 43d is a meandering channel and is connected to the drain D1. The cross-sectional area of the overflow channel 43d is the same as or smaller than the cross-sectional area of the measuring channel 43c.

  The dilution liquid measuring means 44 is disposed downstream of the dilution liquid tank 41 and is connected to each of the first and second dilution tanks 42A and 42B. The diluent metering means 44 includes an introduction channel 44a, a metering channel 44c, and an overflow channel 44d. The introduction flow path 44 a is a flow path for introducing the diluent 40 from the diluent tank 41. A metering flow path 44c and an overflow flow path 44d extend from the introduction flow path 44a via a branching portion 44b. The measuring channel 44c is for temporarily retaining an accurate amount of the diluent 40 in order to dilute the blood to a predetermined concentration. As shown in FIGS. 3 and 4, the metering channel 44 c has a large cross section 44 ca and two tapered portions 44 cb. The large cross section 44c has a width of about 2 mm and a depth of about 2 mm, and a volume of about 50 μL. The two taper portions 44cb are connected to the front and rear ends of the large cross-section portion 44ca, respectively, and the flow of the dilution liquid 40 flows into the large cross-section portion 44ca and flows out of the large cross-section portion 44ca. It is for preventing. As shown in FIGS. 1 and 2, the overflow channel 44d is connected to the drain D2.

  The first and second dilution tanks 42A and 42B are tanks in which blood is diluted. Each of the first and second dilution tanks 42A and 42B has a diameter of about 6 mm and a depth of about 2 mm, and has a volume of 50 μL or more. The first dilution tank 42A is connected to the blood measuring means 43 and the diluent measuring means 44, and the blood measured by the blood measuring means 43 is diluted with the diluent 40 measured by the diluent measuring means 44. It is. The second dilution tank 42B is connected to the first dilution tank 42A and the dilution liquid measuring means 44, and the sample blood diluted in the first dilution tank 42A is diluted with the dilution liquid 40 measured by the dilution liquid measuring means 44. It is a tank to be done. A metering channel 46 is provided between the first dilution tank 42A and the second dilution tank 42B. In the present embodiment, since the dilution ratios in the first and second dilution tanks 42A and 42B are the same, the measurement flow path 46 has the same size as the measurement flow path 43c described above.

  The plurality of analysis units 5A, 5B, 5C, and 5D are parts where analysis of specific components in blood is performed. The first and second analysis units 5A and 5B are analysis units using an electrical resistance detection method, and the first analysis unit 5A is for white blood cells and the second analysis unit 5B is for red blood cells. On the other hand, the third and fourth analysis units 5C and 5D are analysis units using an optical technique, and the third analysis unit 5C is for Hb and the fourth analysis unit is for CRP.

  The first analyzer 5A is connected to the first dilution tank 42A via the buffer tank 45, and is a part for counting white blood cells using the sample blood diluted in the first dilution tank 42A. As shown in FIG. 5 and FIG. 6, the first analysis unit 5 </ b> A has a pore 53 and a pair of electrodes 51 sandwiching the pore 53, and enables counting using an electrical resistance detection method. It is configured. The pore 53 has a narrow width of about 50 μm while the width of the flow path before and after the pore is about 250 μm. This width is determined so that the change in electrical resistance between the pair of electrodes 51 becomes significantly large when white blood cells pass. A pair of electrodes 51 are provided in the flow path portion expanded in a substantially circular shape around the pore 53. The pair of electrodes 51 is made of one or a plurality of types selected from, for example, gold, platinum, palladium, and carbon, and is formed by a printing technique. As shown in FIG. 6, each electrode 51 is electrically connected to the wiring pattern 22 through the through hole 52. The through hole 52 and the wiring pattern 22 are made of, for example, copper.

  The second analysis unit 5B is connected to the second dilution tank 42B and is a part for counting red blood cells using the sample blood diluted in the second time in the second dilution tank 42B. The second analysis unit 5B has substantially the same structure as the first analysis unit described with reference to FIGS.

  The third and fourth analysis units 5C and 5D are connected to the buffer tank 45 independently. As shown in FIGS. 7 and 8, the third and fourth analyzers 5C and 5D have a reflection film 55 provided in the flow path portion enlarged in a substantially circular shape, and each of them is Hb by an optical method. And a site for measuring CRP. The reflection film 55 is made of one or more kinds selected from, for example, gold, platinum, and palladium, and is formed together with the electrode 51 by a printing method. As shown in FIG. 8, a reagent 56 is applied to the upper surface of the enlarged flow path portion in the figure. The reagent 56 is mixed with the sample blood and can measure Hb or CRP by an optical method. In the present embodiment, the third and fourth analyzers 5C and 5D are irradiated with light through the transparent main body 1, and the reflected light is detected to measure Hb and CRP. .

  Flow measurement units 6A and 6B are connected to the first and second analysis units 5A and 5B, respectively. The flow rate measuring units 6A and 6B are parts for measuring the flow rate of the sample blood that has passed through the first and second analyzing units 5A and 5B, respectively, and have meandering channels 61 and a plurality of electrodes 62. . The meandering channel 62 has a sufficient volume while increasing the length in the flow direction. In the present embodiment, the meandering channel 62 serves as a storage means capable of storing at least 50 μL or more of the analyzed sample blood that has passed through the first analysis unit 5A or the second analysis unit 5B. The plurality of electrodes 62 are arranged at a constant pitch in the flow direction of the meandering channel 61. Each electrode 62 has the same structure as the electrode 51 described above.

  Next, blood analysis using the cartridge A will be described below.

  First, in FIG. 1, blood as a sample liquid is introduced from a liquid inlet 3 using a dropper or the like. The cartridge A into which blood has been introduced is loaded into an analyzer (not shown). In this loading, the connector 8 is connected to a connector (not shown) of the analyzer. At this time, the liquid inlet 3 and the drains D1 to D7 shown in FIG. 2 are connected to a plurality of air discharge nozzles or air suction nozzles connected to the pump provided in the analyzer. The analyzer is configured such that the connection state between the pump and the air discharge nozzle and air suction nozzle can be switched as appropriate.

  Next, the blood is measured by the blood measuring means 43. The procedure will be described with reference to FIGS. Since the main body 1 and the printed wiring board 2 have a hydrophobic surface, the blood S introduced from the liquid introduction port 3 does not flow due to capillary action and stays in the liquid introduction port 3. After blood S is introduced, air is discharged from the liquid inlet 3. As a result, as shown in FIG. 9, blood S flows out from the liquid introduction port 3 through the introduction flow path 43a to the measurement flow path 43c and the overflow flow path 43d. Since the measurement flow path 43c and the overflow flow path 43d have substantially the same cross-sectional area, the pressure loss resistance when the blood S flows is also substantially the same. Accordingly, the blood S flows so that the lengths in the flow direction of the blood S included in the measurement flow path 43c and the overflow flow path 43d are substantially the same.

  If the above discharge is continued, as shown in FIG. 10, the blood flow S is filled with the blood S as shown in FIG. At this time, blood S is present in the overflow channel 43d by a length corresponding to the measuring channel 43c.

  If the discharge is further continued from the state shown in FIG. 10, the state shown in FIG. 11 is obtained. That is, since the orifice 43e is provided on the downstream side of the measuring channel 43c, the pressure loss resistance when the blood S flows is very large. On the other hand, since the overflow channel 43d has a uniform cross section in the flow direction, the pressure loss resistance is significantly smaller than that of the orifice 43e. As a result, the blood S continues to flow in the overflow channel 43d while the blood S remains in the measurement channel 43c.

  When the discharge is further continued, all the blood S flows out from the liquid introduction port 3, and the state shown in FIG. In this figure, as a result of the continuation of the above discharge, air has entered the upstream portion of the introduction flow path 43a and the overflow flow path 43d instead of the blood S, and the blood Sa staying in the measurement flow path 43c is retained. Separated from blood S. Since the amount of blood S injected into the liquid inlet 3 is substantially constant, the time from the start of air discharge shown in FIG. 9 to the state shown in FIG. 12 is substantially constant. This fixed time is measured by a timer provided in the analyzer, and the discharge is stopped after a fixed time has elapsed.

  Then, as shown in FIG. 13, when air is discharged from the liquid inlet 3 again after the drain D1 is closed by the analyzer, the blood Sa staying in the measuring flow path 43c passes through the orifice 43e to the second position. It flows out to 1 dilution tank 42A. With the above procedure, measurement of a predetermined amount of blood Sa is completed, and about 0.5 μL of blood Sa, which is a predetermined amount, stays in the first dilution tank 42A.

  Next, the diluent 40 is weighed by the diluent metering means 44. The procedure will be described with reference to FIGS. FIG. 14 shows a state in which the measurement of the diluent 40 is started. To achieve this state, for example, an aluminum pack (not shown) in which the diluent 40 is enclosed in the diluent tank 41 is ruptured so that the diluent 40 can flow out. Since the main body 1 and the printed wiring board 2 have a hydrophobic surface, even if the aluminum pack is ruptured, the diluted solution 40 does not flow unduly due to a capillary phenomenon or the like.

  After allowing the diluent 40 to flow out, the drain D2 is closed and the drain D3 is opened, and the discharge of air from the diluent tank 41 is started. As a result, as shown in FIG. 15, the diluent 40 is pushed out of the diluent tank 41 and flows into the metering channel 44c through the introduction channel 44a.

  When the above discharge is further continued, the state shown in FIG. 16 is obtained. A large cross section 44ca is formed on the downstream side of the taper portion 44cb in the measuring channel 44c. As described above, since the surface forming the large cross-sectional view 44ca is a hydrophobic surface, the dilution liquid 40 is subjected to a surface tension that stays upward in the drawing and does not progress due to capillary action. The propulsive force by the discharge gradually sends the diluent 40 downward in the figure against the surface tension. When the discharge is continued as it is, as shown in FIG. 17, the inside of the measurement flow path 44 c is filled with the diluent 40, and the diluent 40 moves toward the drain D <b> 3. It is detected, for example, by an electric resistance means using an electrode (not shown) or an optical means using a reflective film (not shown) that the tip of the diluent 40 has reached before the drain D3. The discharge is stopped with this detection.

  Then, as shown in FIG. 18, for example, after drain D3 and drain D7 shown in FIG. 1 are closed, air is discharged from, for example, drain D2 shown in FIG. 18, or downstream of first dilution tank 42A. Aspirate from any drain located in the As a result, the diluent 40 staying in the measuring channel 44c is sent out to the first dilution tank 42A. The measurement of the diluent 40 is completed by the above procedure, and about 50 μL of the diluent 40a which is a predetermined amount stays in the first dilution tank 42A.

  Thereafter, in the first dilution tank 42A, about 0.5 μL of blood Sa and about 50 μL of diluted solution 40a are mixed to obtain specimen blood as a diluted sample solution diluted 100 times. For this mixing, a stirring bar 42Aa built in the first dilution tank 42A is used. The stirrer 42Aa is formed by sealing a small piece made of a ferromagnetic material such as iron with a fluorine resin such as Teflon (registered trademark). The stirrer 42Aa is rotatable by a magnetic force generation source provided in the analyzer in which the cartridge A is loaded. By this rotation, blood Sa and diluent 40a are mixed. In FIG. 19, blood Sa and dilution liquid 40a are omitted. The dilution according to the above procedure will be referred to as the first dilution.

  After the first dilution in the first dilution tank 42A is completed, the white blood cell count by the first analysis unit 5A and the Hb and CRP analysis by the third and fourth analysis units 5C and 5D are performed. As shown in FIG. 1, a buffer tank 45 is connected to the first dilution tank 42A. The sample blood diluted 100 times is sent to the buffer tank 45.

  A procedure for counting white blood cells by the first analyzer 5A using a part of the sample blood stored in the buffer tank 45 will be described with reference to FIGS. For this counting, the first analysis unit 5A and the first flow rate measurement unit 6A provided on the downstream side thereof are used. FIG. 19 shows a state in which the white blood cell count is started, and the specimen blood DS as a diluted sample liquid diluted 100 times remains in the buffer tank 45. As shown in FIG. 20, a dry hemolyzing agent 57 is applied to the buffer tank 45. The dry hemolyzing agent 57 is for performing a hemolysis process for destroying red blood cells in the specimen blood. This hemolysis treatment is performed in order to eliminate the influence of red blood cells on the white blood cell coefficient. In this state, for example, air suction is started from the drain D4. Then, as shown in FIG. 21, the sample blood DS flows out from the buffer tank 45 and flows through the first analysis unit 5A.

  When the suction from the drain D4 is further continued, the tip portion of the sample blood DS reaches the electrode 62a located on the most upstream side among the plurality of electrodes 62 as shown in FIG. For example, by monitoring the conduction between the electrode 62a and the electrode 51, it is possible to detect that the tip portion of the sample blood DS has reached the electrode 62a. Using this detection as a guide, counting of white blood cells by the first analyzer 5A is started. As described above, since the pore 53 is narrow, when white blood cells pass through, the electrical resistance between the pair of electrodes 51 increases momentarily. As a result, when the electrical resistance between the pair of electrodes 51 is monitored in time series, a pulse signal is generated corresponding to the passage of white blood cells. The number of pulse signals is integrated.

  When the aspiration is continued while integrating the pulse signals, the tip portion of the sample blood DS reaches the second electrode 62b counted from the upstream side among the plurality of electrodes 62 as shown in FIG. This arrival can be detected, for example, by monitoring conduction between the electrodes 62a and 62b. The flow rate of the sample blood DS that has passed through the first analyzer 5A during the period from when the tip of the sample blood DS reaches the electrode 62a until it reaches the electrode 62b is the sample blood DS that can stay between the electrodes 62a and 62b. Is the same amount. Since the flow direction distance between the electrodes 62a and 62b is known, the flow rate of the sample blood DS that has passed through the first analyzer 5A can be known. The number of white blood cells per unit volume of the sample blood DS is obtained from this flow rate and the number of pulses integrated. Thereby, the number of white blood cells per unit volume of blood S can be counted.

  Thereafter, it is possible to further improve the accuracy of counting by continuing the above suction and repeating counting. In the present embodiment, the first flow rate measuring unit 6 </ b> A includes a large number of electrodes 62. Therefore, if the number of pulses is stored each time the tip of the sample blood DS reaches each electrode 62 after the electrodes 62a and 62b, a large number of counts are possible. Since this is synonymous with counting using a larger amount of sample blood DS, the counting accuracy can be improved. Then, for example, as shown in FIG. 24, when it is detected that the distal end portion of the sample blood DS has reached the electrode 62n located on the most downstream side among the plurality of electrodes 62, the counting process by the first analyzer 5A is performed. Can be terminated. Further, as is clear from this figure, when the counting by the first analyzer 5A is completed, the analyzed sample blood DS is in a state of staying in the meandering flow path 61.

  On the other hand, the analysis by the third and fourth analysis units 5C and 5D is performed by, for example, sucking from the drains D5 and D6 after the counting by the first analysis unit 5A is finished, and the sample blood DS is collected by the third and fourth analysis units This is performed by reaching the reflective films 55 of 5C and 5D. At this time, as shown in FIG. 8, the sample blood DS reacts with the reagent 56 and becomes in a state where each of Hb and CRP can be analyzed. In this state, light is emitted from the analyzer through the main body 1 to the respective reflective films 55, and the reflected light is received through the main body 1 by a light receiving element or the like provided in the analyzer. By appropriately processing this light, Hb and CRP can be analyzed. Unlike the present embodiment, the printed wiring board 2 may be replaced with a substrate made of a transparent material. In this case, the reflective film 55 is unnecessary. Each of the third and fourth analysis units 5C and 5D has a structure sandwiched between the transparent main body 1 and the substrate. Therefore, it is possible to analyze Hb and CRP by so-called transmission measurement.

  Next, the procedure for counting red blood cells by the second analyzer 5B will be described below. Prior to this counting, the second dilution is performed by the dilution means 4 shown in FIG. The procedure for the second dilution is similar to the procedure for the first dilution described with reference to FIGS. That is, in the first dilution, the blood S was diluted about 100 times with the diluent 40, whereas in the second dilution, the blood S was 100 times obtained by the first dilution. The diluted specimen blood DS is further diluted by about 100 times using the diluent 40. The sample blood thus obtained corresponds to the blood S diluted 10,000 times. With the 100-fold diluted sample blood DS remaining in the first dilution tank 42A, about 50 μL of the sample blood DS is sent to the second dilution tank 42B using the measurement channel 46 shown in FIG. The measurement of the sample blood DS using the measurement channel 46 is substantially the same as the measurement procedure described with reference to FIGS. On the other hand, in the measurement by the diluent measuring means 44 described with reference to FIGS. 14 to 18, for example, the drains D1, D3, D4, D5, D6, and D8 shown in FIG. 1 are closed and the drain D2 is opened. Above, suction from the drain D7. Thereby, about 50 μL of the diluent 40 can be delivered to the second dilution tank 42B. In the second dilution tank 42B, the dilution is substantially 10,000 times with 5 μL of the sample blood DS and about 50 μL of the diluent 40. In this dilution, the mixing of the sample blood DS and the diluent 40 is promoted by rotating the stirrer 42Ba by a magnetic force.

  Using the 10,000-fold diluted sample blood obtained by the above procedure, red blood cells are counted by the second analyzer 5B. This counting procedure is almost the same as the counting procedure by the first analyzer 5A. The point of measuring the flow rate using the second flow rate measurement unit 6B is the same as the flow rate measurement using the first flow rate measurement unit 6A.

  Next, the operation of the cartridge A will be described.

  According to the present embodiment, for example, the rotation member having the measurement flow path needs to be rotated for the measurement of the blood S by the blood measurement means 43, the measurement of the diluent 40 by the dilution liquid measurement means 44, and the measurement of the sample blood DS There is no. This is because the blood measuring means 43 and the diluent measuring means 44 are constituted by flow paths configured as so-called T-shaped paths. Manufacturing the blood measuring means 43 and the diluent measuring means 44 is simpler than making the rotating member, and the manufacturing efficiency can be improved. In the measuring step, it is not necessary to rotate the rotating member or the like, and there is no risk of liquid leakage. Therefore, the cartridge A can be made as a so-called disposable type, which is relatively easy to manufacture and can be used in an appropriate sanitary state.

  Moreover, according to the blood measurement means 43 and the dilution liquid measurement means 44, the blood S, the dilution liquid 40, and the sample blood DS can be accurately measured. Therefore, the accuracy of analysis in the first to fourth analysis units 5A, 5B, 5C, and 5D can be increased.

  In the first dilution tank 42A and the second dilution tank 42B, the stirrers 42Aa and 42Ba are rotated. Thereby, for example, as shown in FIG. 18, it is possible to physically stir the blood Sa and the diluent 40a in the first dilution tank 42A, and these can be sufficiently mixed. Similarly, the sample blood DS and the diluent 40a can be reliably mixed in the second dilution tank 42B. Therefore, the first to fourth analysis units 5A, 5B, 5C, and 5D are suitable for improving the accuracy of analysis.

  By using the dry hemolyzing agent 57 applied to the buffer tank 45, it is possible to appropriately eliminate the influence of red blood cells. For example, it is possible to accurately count white blood cells. Further, the dried hemolyzing agent 57 itself is solid and hardly contains moisture. Therefore, the cartridge A before use can be in a dry state. This is preferable for reducing the weight of the cartridge A and improving the sanitary condition.

  Further, according to the present embodiment, two-stage dilution can be performed using the first and second dilution tanks 42A and 42B. Therefore, two types of dilution with relatively high magnification, for example, 100-fold dilution and 10,000-fold dilution, are possible. As a result, it is possible to collectively perform the analysis of significantly different dilution ratios suitable for each of the white blood cell count and the red blood cell count. Further, by providing the blood measuring means 43 and the diluent measuring means 44, it is possible to dilute with a sufficiently accurate dilution factor. Weighing using the large cross section 44ca is particularly effective for high magnification dilution.

  The flow rate measurement using the first and second flow rate measurement units 6A and 6B is very simple and accurate. This not only enables accurate counting of red blood cells and white blood cells, for example, but also eliminates the need for a mechanism for realizing a constant flow rate in the analyzer. Therefore, it is advantageous for simplification of the analyzer.

  By using the printed wiring board 2 provided with the through holes 52, portions other than the electrodes 51 and 62 can be smooth surfaces. This is suitable for bonding the main body 1 and the printed wiring board 2 in a liquid-tight manner.

  FIG. 25 is a partially enlarged view showing a modified example of the cartridge according to the present invention. In this modification, the main body 1 and the lead 54 are integrally formed by insert molding. One end of the lead 54 exposed in the flow path is the electrode 51 described above. The other end of the lead 54 is exposed from the main body 1 and constitutes the connector 8 shown in FIGS. 1 and 2. According to such a configuration, the main body 1 and the electrode 51 can be formed in a lump without going through a dedicated printing process for forming the electrode 51, which is suitable for improving the manufacturing efficiency. Yes.

  The cartridge according to the present invention is not limited to the above-described embodiment. The specific configuration of each part of the cartridge according to the present invention can be changed in various ways.

  The material of the main body is not limited to a transparent material and may be partially opaque. In this case, at least a portion corresponding to the optical analysis portion is a transparent portion. If a printed wiring board is used, it is preferable for thinning, but a so-called rigid board may be used. As the diluted sample liquid detection means, in addition to the one including an electrode, for example, an optical means may be used.

  The dilution factor in the diluting means can be further increased by appropriately setting the size of the flow path and the like. Moreover, it is not limited to two-stage dilution, For example, it is good also as a structure which performs the dilution of only 1 time, or the dilution 3 times or more.

  The cartridge according to the present invention is not limited to the counting of blood and can be used for analysis of various sample solutions. In addition, the cartridge according to the present invention does not include an analysis unit, and may be configured to be used only for preparing a diluted sample solution for a blood cell coefficient, for example.

It is a whole top view which shows an example of the cartridge which concerns on this invention. It is a whole perspective view which shows an example of the cartridge which concerns on this invention. It is a principal part top view which shows the measurement flow path of an example of the cartridge which concerns on this invention. It is principal part sectional drawing in alignment with the IV-IV line of FIG. It is a principal part top view which shows the electrical resistance type | formula analysis part of an example of the cartridge which concerns on this invention. It is principal part sectional drawing in alignment with the VI-VI line of FIG. It is a principal part top view which shows the optical analysis part of an example of the cartridge which concerns on this invention. It is principal part sectional drawing which follows the VIII-VIII line of FIG. In the blood measurement procedure using an example of the cartridge according to the present invention, it is a main part plan view showing a blood introduction state. In the blood measurement procedure using an example of the cartridge concerning the present invention, it is a principal part top view showing the state where the measurement channel was filled up. FIG. 6 is a plan view of a principal part showing continuous inflow into an overflow channel in a blood measurement procedure using an example of a cartridge according to the present invention. In the blood measurement procedure using an example of the cartridge concerning the present invention, it is a principal part top view showing the state where blood was separated in the measurement channel. In the blood measurement procedure using an example of the cartridge concerning the present invention, it is a principal part top view showing the state where blood was sent out to the 1st dilution tank. FIG. 7 is a plan view of a principal part showing a starting state in a diluent measurement procedure using an example of a cartridge according to the present invention. FIG. 10 is a plan view of a principal part showing a diluent introduction state in a diluent measurement procedure using an example of the cartridge according to the present invention. FIG. 6 is a plan view of a principal part showing continuous inflow into a measurement flow path in a diluent measurement procedure using an example of a cartridge according to the present invention. FIG. 7 is a plan view of a principal part showing a state in which a metering channel is filled in a diluent measuring procedure using an example of a cartridge according to the present invention. FIG. 7 is a plan view of a principal part showing a state in which a diluent is delivered to a first dilution tank in a diluent measurement procedure using an example of a cartridge according to the present invention. In the blood cell count procedure using an example of the cartridge concerning the present invention, it is a principal part top view showing the initial state. It is principal part sectional drawing which follows the XX-XX line of FIG. In the blood cell counting procedure using an example of the cartridge according to the present invention, it is a principal plan view showing a state in which the sample blood has reached the first analysis unit. In the blood cell counting procedure using an example of the cartridge according to the present invention, FIG. In the blood cell counting procedure using an example of the cartridge according to the present invention, it is a principal plan view showing a state in which the tip portion of the sample blood has reached the second electrode from the upstream. In the blood cell counting procedure using an example of the cartridge according to the present invention, it is a main part plan view showing a state where the tip portion of the sample blood has reached the most downstream electrode. It is principal part sectional drawing which shows the modification of the cartridge which concerns on this invention. It is a whole perspective view which shows an example of the conventional cartridge.

Explanation of symbols

A Cartridge D1-D8 Drain DS Sample blood (diluted sample solution)
S blood (sample liquid)
Sa blood (separated sample liquid)
DESCRIPTION OF SYMBOLS 1 Main body 2 Printed wiring board 3 Liquid inlet 4 Dilution means 5A 1st analysis part (electrical resistance type analysis part)
5B 2nd analysis part (electric resistance type analysis part)
5C 3rd analysis unit (optical analysis unit)
5D Fourth analysis unit (optical analysis unit)
6A 1st flow measurement part 6B 2nd flow measurement part 8 Connector 22 Wiring 40 Dilution liquid 40a Separated dilution liquid 41 Dilution liquid tank 42A 1st dilution tank 42Aa Stirrer 42B 2nd dilution tank 42Ba Stirrer 43 Blood measuring means ( Sample liquid measuring means)
43a Introducing channel 43b Branching part 43c Metering channel 43d Overflow channel 43e Orifice 44 Diluent metering means 44a Introducing channel 44b Branching part 44c Metering channel 44ca Large cross section 44cb Taper part 44d Overflow channel 45 Buffer tank 46 Metering flow Path 51 electrode 52 through hole 53 pore 54 lead 55 reflective film 56 reagent 57 dry hemolyzing agent 61 meandering channel (storage means)
62 Electrode (Diluted sample solution detection means)

Claims (24)

A liquid inlet through which the sample liquid is introduced;
A diluting liquid tank in which a diluting liquid for diluting the sample liquid is stored, a sample liquid measuring means for separating a predetermined amount from the sample liquid introduced from the liquid introducing port, and the sample liquid and the diluting liquid One or more dilution tanks for mixing at least a portion, and a dilution means comprising:
A cartridge to be loaded into a separate device,
The sample liquid metering means includes an introduction channel extending from the liquid introduction port, a metering channel and an overflow channel connected to the introduction channel via a branch portion,
The cartridge, wherein the metering channel is directed to the dilution tank.   The cartridge according to claim 1, wherein a stirring bar is accommodated in the dilution tank.   The cartridge according to claim 1, wherein an orifice is interposed between the measuring channel and the dilution tank.   4. The portion of the overflow channel corresponding to at least the length of the metering channel from the branch portion has a cross-sectional area equal to or smaller than a cross-sectional area of the metering channel. Cartridge.   The cartridge according to any one of claims 1 to 4, wherein the diluting means includes diluting liquid measuring means for separating a predetermined amount from the diluting liquid in the diluting liquid tank.   The cartridge according to claim 5, wherein the dilution liquid measuring means includes a measuring flow path having a large cross section and a pair of tapered portions connected to both ends of the large cross section in the flow direction. The dilution means includes first and second dilution tanks,
The first dilution tank is connected to a flow path into which the sample liquid introduced from the liquid introduction port flows and a flow path into which the dilution liquid flows from the dilution liquid tank.
The flow path into which the diluted sample liquid diluted in the first dilution tank flows in and the flow path into which the diluent flows from the dilution liquid tank are connected to the second dilution tank. 7. The cartridge according to any one of 6. A buffer tank disposed on the downstream side with respect to the first dilution tank;
The cartridge according to claim 7, wherein a dry hemolyzing agent is applied to the buffer tank.   Analysis loaded in an analyzer for analyzing the specific component contained in the sample solution by providing one or more analysis units for analyzing the specific component contained in the diluted sample solution diluted by the dilution means 9. The cartridge according to claim 1, wherein the cartridge is configured as a device cartridge.   The cartridge according to claim 9, further comprising one or more storage means for storing the analyzed diluted sample solution. A first analysis unit for analyzing the diluted sample solution diluted in the first dilution tank;
11. The cartridge according to claim 9, further comprising a second analysis unit configured to analyze the diluted sample solution diluted in the second dilution tank.   The cartridge according to claim 9, further comprising a flow rate measurement unit for measuring a flow rate of the diluted sample solution that has passed through the analysis unit.   The cartridge according to claim 12, wherein the flow rate measurement unit includes a meandering flow path and two or more diluted sample liquid detection means arranged at positions separated from each other in the flow direction of the meandering flow path.   The cartridge according to claim 13, wherein the diluted sample solution detection means includes an electrode.   The cartridge according to claim 13 or 14, wherein the meandering flow path serves as the storage means.   The cartridge according to any one of claims 9 to 15, wherein the analysis unit includes an electrical resistance analysis unit having a pore and a pair of electrodes spaced apart from each other with the pore interposed therebetween.   The said analysis part further contains an optical analysis part which has a reflecting film, a translucent part, and the reagent apply | coated to the said reflecting film or the said light projection part, The Claim 9 thru | or 16 cartridge.   The cartridge according to any one of claims 1 to 17, wherein the channel through which the sample solution and the diluted sample solution are flowed is configured by a hydrophobic surface having a contact angle of water of 60 degrees or more.   The cartridge according to claim 18, wherein the flow path includes one having a width / depth of 1 or more and 5 or less. A main body and a printed circuit board pasted on the main body,
The main body is formed with a plurality of recesses or grooves,
The cartridge according to claim 1, wherein a plurality of flow paths or tanks are configured by the printed wiring board covering the plurality of recesses or grooves. A body having a plurality of recesses or grooves,
An electrode formed integrally with the main body by insert molding so as to be exposed in the plurality of recesses or grooves,
A covering member bonded to the main body,
The cartridge according to any one of claims 1 to 19, wherein a plurality of flow paths or tanks are configured by covering the plurality of recesses or grooves with the covering member.   The cartridge according to any one of claims 1 to 21, wherein the sample solution is blood.   The cartridge according to claim 22, wherein the specific component is blood cells such as red blood cells, white blood cells, and platelets.   The cartridge according to claim 22, wherein the specific component is hemoglobin or C-reactive protein.

Images