WO2014122491A1 - Unité de test pour la détermination des caractéristiques physiques d'échantillons contenant des composants liquides - Google Patents

Unité de test pour la détermination des caractéristiques physiques d'échantillons contenant des composants liquides Download PDF

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Publication number
WO2014122491A1
WO2014122491A1 PCT/HU2014/000013 HU2014000013W WO2014122491A1 WO 2014122491 A1 WO2014122491 A1 WO 2014122491A1 HU 2014000013 W HU2014000013 W HU 2014000013W WO 2014122491 A1 WO2014122491 A1 WO 2014122491A1
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WO
WIPO (PCT)
Prior art keywords
flow cuvette
measuring
opening
diverting
current
Prior art date
Application number
PCT/HU2014/000013
Other languages
English (en)
Inventor
Jenő CSIKÓS
Péter Kovács
Bálint Tibor MENDELE
László ORBÁN
László SÜLE
Peter TÓTH-MIKLÓS
Attila Zsolt Tremmel
Gabor VÁRNAGY
Original Assignee
Norma Instruments Zrt.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Norma Instruments Zrt. filed Critical Norma Instruments Zrt.
Publication of WO2014122491A1 publication Critical patent/WO2014122491A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N15/1484—Electro-optical investigation, e.g. flow cytometers microstructural devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • G01N15/134—
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0645—Electrodes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0864—Configuration of multiple channels and/or chambers in a single devices comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G01N2015/135—
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Electro-optical investigation, e.g. flow cytometers
    • G01N2015/1486—Counting the particles

Definitions

  • the subject of the invention relates to a testing unit for determining the physical characteristics of samples containing liquid components, especially for determining the number of blood cells in blood, which has a basic body with a delimiting shell enclosing a flow cuvette and having an upper surface, a lower surface and side surfaces in between the upper surface and the lower surface, where there are at least two current-conducting electrodes located on the inner side of the delimiting shell surrounding the flow cuvette separated at a distance, furthermore, the flow cuvette has an inlet passage serving for inputting the sample to be tested and an outlet passage serving for releasing the tested sample, and there is a measuring gate inside the flow cuvette.
  • microfluidic measurement units may also be learned of, among others, in publication document number WO 201 1/143075, in patent specification registration number EP 2.523.004 as well as in publication document number US 2012/021 1373. Patent specification registration number EP 2.523.004 presents a testing unit suitable for performing optical measurements while publication document number US 2012/021 1373 presents a testing unit based on the measurement of impedance.
  • the undoubted advantage of the applied solutions is that the smaller dimensions create the possibility for moving the equipment more easily, and the use of the sample and reagents used for performing the measurements can be greatly reduced.
  • the disadvantage of measurements made on the basis of the volumetric impedance method is that the direct current needed for measurements with the given method on the electrodes for electricity feed necessarily generates the so-called "redox reaction", as a result of which gases form on the surfaces of the electrodes.
  • the bubbles forming in this way disturb measuring, they may cause noise, or in extreme cases they may even terminate the liquid-electrode contact, which may make measuring impossible.
  • Our aim with the solution according to the invention was to overcome the deficiencies of testing units suitable for making measurements based on the known volumetric impedance method and to create a version that eliminates the measuring problems deriving from bubble formation and thus results in higher reliability and makes greater measurement precision possible.
  • the solution according to the invention is based on the recognition that if the flow cuvette containing the measuring point is created with a geometrical construction unlike the ordinary, in such a way that one of the current- conducting electrodes situated before the measuring point is placed in a controlled-flow channel in a way different from the known methods, and in this channel a flow moving away from the measuring point is created with novel liquid-flow control, then the bubbles forming before the measuring point can be prevented from getting to the measuring point, so their disturbing effect cannot be asserted there and so the task can be solved.
  • the testing unit for determining the physical characteristics of samples containing liquid components, especially for determining the number of blood cells in blood, - which has a basic body with a delimiting shell enclosing a flow cuvette and having an upper surface, a lower surface and side surfaces in between the upper surface and the lower surface, where there are at least two current-conducting electrodes located on the inner side of the delimiting shell surrounding the flow cuvette separated at a distance, furthermore, the flow cuvette has an inlet passage serving for inputting the sample to be tested and an outlet passage serving for releasing the tested sample, and there is a measuring gate inside the flow cuvette, - is set up in such a way that the basic body is supplemented with a pressure channel suitable for introducing the diverting medium and a discharge channel suitable for discharging the diverting medium, furthermore, there is a diverting opening and an outlet opening in the section of the flow cuvette between the inlet passage and the outlet passage, the pressure channel suitable for introducing the diverting medium
  • the current-conducting electrodes are formed by vapour-depositing extensive metal conducting layers, e.g. platinum layers, applied onto the upper surface and/or the lower surface of the delimiting shell, furthermore, the first measuring sensor and the second measuring sensor of the electric measuring body are formed by vapour-depositing extensive metal conducting layers, e.g. platinum layers, applied onto the upper surface and/or the lower surface of the delimiting shell.
  • extensive metal conducting layers e.g. platinum layers
  • the section of the flow cuvette between the first current- conducting electrode and the second current-conducting electrode is at least partly filled with an electricity-conducting medium, and so the first current-conducting electrode and the second current-conducting electrode are in an electrically conducting connection via the electricity-conducting medium.
  • the distance between the upper surface and the lower surface of the delimiting shell of the basic body in contact with the flow cuvette is between 10- 100 microns in at least a part of the measuring gate of the flow cuvette, while the distance between the first side surface and the second side surface of the delimiting shell of the basic body in contact with the flow cuvette is between 10-100 microns in at least a part of the measuring gate of the flow cuvette.
  • the testing unit in the part of the flow cuvette in the environment of the measuring gate and/or in the measuring gate there is an optical sensor part-unit.
  • the testing unit according to the invention has numerous advantageous characteristics. The most important of these is that as a consequence of the unique setup of the flow cuvette there is no bubble formation in the environment of the measuring sensors of the electric measuring body situated at the measuring gate, and so the bubbles cannot cause measuring faults, as a result of which the reliability of the measuring result and the measuring precision both increase significantly.
  • testing unit according to the invention contains fewer moving parts, the liquid pathways to be travelled by the sample are significantly shortened and so the measurements may be performed with a low amount of energy use, and with a minimal sample and reagent demand.
  • a further advantage deriving from this is the further increase in reliability of the testing unit, the reduction of maintenance demand and the reduction of fault probability, which, together, may result in a great increase in the efficiency of the testing unit according to the invention and in a significant decrease in the specific testing expenses as compared with traditional devices.
  • testing unit may be operated without an optical head in a three-part machine, while in a five-part machine it may be operated with an optical head, in other words its area of use may be extended. Furthermore, the use of the testing unit according to the invention makes it possible to perform simultaneous volumetric impedance and optical measurements, which may result in the more favourable and more reliable evaluation of the test results.
  • the testing unit according to the invention may be used to create a bedside, in vitro diagnostic device which is based on a measuring procedure that may be performed using the traditional and medically accepted volumetric impedance method, and which is capable of producing fast and precise results in the hospital ward, which is a fundamental requirement in the case of urgent tests and which the known solutions were unable to comply with in the desired way.
  • Figure 1 shows a top view of the testing unit according to the invention, in partial cross-section
  • Figure 2 shows the view of the testing unit according to figure 1 from the direction II
  • Figure 3 shows a view picture of the testing unit according to the invention, in partial cross-section.
  • FIG 1 and in figure 2 an embodiment of the testing unit according to the invention can be seen, which can be used favourably in the measuring head of blood cell counting devices.
  • the basic body 10 here essentially consists of three pieces, a cover plate 2, a base plate 3 and an internal plate 4. This structure makes it easier to precisely realise the basic body 10, to set up the flow cuvette 20 consisting of small sized passages.
  • the cover plate 2, the base plate 3 and the internal plate 4 are made from a material that has good resistance to the material of the samples and the reagents, e.g. from glass or from plastic with a suitable composition, and they are built together into the basic body 10 using known procedures for producing microfluidic chips.
  • Figure 2 also well illustrates that the delimiting shell 1 1 surrounding the flow cuvette 20 is established between the cover plate 2, the base plate 3 and the internal pate 4 of the basic body 10, which is formed by the upper surface 1 la from the direction of the cover plate 2, by the lower surface 1 lb from the direction of the base plate 3 and by the first side surface 1 1 c and the second side surface 1 Id in the internal plate 4.
  • the distance "t" between the upper surface 1 la of the cover plate 2 and the lower surface 1 lb of the base plate 3 falls within the range of 10-100 microns, in this case the distance "t" is 100 microns.
  • FIG 1 it shows the top view of the testing unit.
  • the flow cuvette 20 surrounded by the delimiting shell 1 1 of the basic body 10 is interrupted with a diverting opening 22 and an outlet opening 23 between the inlet passage 21 and the outlet passage, on one of the side surfaces 1 1c of the delimiting shell 1 1.
  • the pressure channel 12 opens into the diverting opening 22, and in relation to the flow cuvette 20 it is situated so that the main axis 12a of the pressure channel 12 and the main progress direction l a of the sample 1 form an acute-angled angle of obliquity "a", when viewed from the direction of the inlet passage 21.
  • the discharge channel 13 starts from the outlet opening 23, and when viewed from the inlet passage 21 , its main axis 13a and the main progress direction la of the sample 1 form an obtuse-angled cone angle " ⁇ ".
  • This geometrical construction makes it possible to forward the diverting medium 40 through the pressure channel 12 into the flow cuvette 20 so that then it can flow out from the flow cuvette 20 through the discharge channel 13.
  • this diverting medium 40 is situated in the pressure channel 12, in a part of a section of the flow cuvette 20 between the diverting opening 22 and the outlet opening 23, and basically in the discharge channel 13.
  • the measuring gate 30 is situated on the section of the flow cuvette 20 near the outlet passage 24, and the spacing "k" between the measuring gate 30 and the first side surface 1 1c and the second side surface l id can be 10-100 microns, in the present case the spacing k" is 50 microns.
  • the first measuring sensor 31a is situated on the side of the measuring gate 30 facing the input passage 21, while the second measuring sensor 31b is situated on the side of the measuring gate 30 facing the output passage 24.
  • the first measuring sensor 31a and the second measuring sensor 31 b are vapour-depositing platinum layers applied onto the lower surface 1 lb of the delimiting shell 1 1 on the base plate 3 of the basic body 10.
  • first current-conducting electrode 60 is situated near the outlet passage 24 of the flow cuvette 20, but after the second measuring sensor 31b of the electric measuring body 31 of the measuring gate 30, while the second current-conducting electrode 70 is situated at a "T" interval from the first current- conducting electrode 60, in the discharge channel 13, further away from the outlet opening 23 of the discharge channel 13, in the basic body 10.
  • first current-conducting electrode 60 and the second current-conducting electrode 70 are vapour-depositing platinum layers applied onto the lower surface 1 lb of the delimiting shell 1 1.
  • first measuring sensor 31a and the second measuring sensor 31b, and the first current-conducting electrode 60 and the second current-conducting electrode 70 may also be situated on the upper surface 1 1a of the delimiting shell 1 1 , not only on the lower surface 1 lb of the delimiting shell, or even on both of them.
  • the section of the flow cuvette 20 and the discharge channel 13 between the first current-conducting electrode 60 and the second current-conducting electrode 70 is filled with electricity- conducting medium 50.
  • This electricity-conducting medium 50 may be, for example, a material contained in and mixed with the sample 1, but even the diverting medium 40 itself may have favourable electricity-conducting characteristics.
  • This electricity- conducting medium 50 makes it possible to create an electric space in the environment of the measuring gate 30, which the components of the sample 1 can change while passing through the measuring gate 30, and which change can be detected by the electric measuring body 31 of the measuring gate 30.
  • Figure 3 basically shows a view picture of the basic body 10 of the testing unit shown in figure 1 and figure 2, after the cover plate 2 has been partly removed.
  • the flow cuvette 20 situated between the inlet passage 21 and the outlet passage, surrounded by the delimiting shell 1 1, and the pressure channel 12 joins its diverting opening 22, and the discharge channel 13 leads out from its output opening 23, and near the output passage 24 there is the measuring gate 30.
  • the testing unit takes place in the following way.
  • the tested sample 1 enters the flow cuvette 20 surrounded by the delimiting shell 1 1.
  • the sample 1 mixed with the electricity-conducting medium 50 gradually progresses from the inlet passage 21 towards the outlet passage 24 following the main progress direction l a.
  • the diverting medium 40 flows towards the diverting opening 22, and on reaching it the diverting medium 40 also enters the flow cuvette 20, where it progresses along the first side surface 1 1c of the delimiting shell 1 1 towards the outlet passage 23.
  • the flow of the mediums in the flow cuvette 20 is laminar, so the diverting medium 40 entering the flow cuvette 20 through the diverting opening 22 does not get mixed together with the sample 1 , but in the section of the flow cuvette 20 after the diverting opening 22 the sample mixed with the electricity conducting medium and the diverting medium progress side-by-side and fill the flow cuvette 20 surrounded by the delimiting shell.
  • the bubbles formed in the discharge channel 13 cannot reach the measuring gate 30.
  • the bubbles formed on the first current-conducting electrode 60 are carried away by the sample 1 and the electricity-conducting medium 50 that have passed through the measuring gate 30 and are removed from the flow cuvette 20 through the outlet passage 24, so the bubbles formed on the first current-conducting - I I -
  • the first measuring sensor 31a and second measuring sensor 31b of the electric measuring body 31 of the measuring gate 30 "monitor" the electric space between the two of them, and when any component of the sample 1 arrives between the first measuring sensor 31a and the second measuring sensor 31b and changes the impedance of the electric space between the first measuring sensor 31 a and the second measuring sensor 31b, it senses this and sends a signal to the evaluating and processing part-unit not shown in the figures.
  • any component of the sample 1 just passes through the measuring gate 30 it is also sensed by the optical sensor part-unit 80, which also sends a signal to the evaluating and processing part-unit.
  • the testing unit according to the invention can be used favourably in the measuring head of devices that count blood cells, but it is also suitable for use in general particle- counting devices, if the aim is to provide reliable and accurate measurement results.
  • sample 1 a main progress direction cover plate base plate internal plate 0 basic body 1 1 delimiting shell

Abstract

La présente invention concerne une unité de test pour la détermination des caractéristiques physiques d'échantillons contenant des composants liquides, en particulier pour la détermination du nombre de cellules sanguines dans du sang, qui comprend un corps de base (10) doté d'une enveloppe délimitante (11) entourant une cuvette d'écoulement (20) et ayant une surface supérieure (11a), une surface inférieure (11b) et des surfaces latérales (11c, 11d) entre la surface supérieure (11a) et la surface inférieure (11b), au moins deux électrodes conductrices de courant (60, 70) étant situées sur le côté interne de l'enveloppe délimitante (11) entourant la cuvette d'écoulement (20), séparées à une distance (T), et en outre, la cuvette d'écoulement (20) ayant un passage d'entrée (21) servant à l'introduction de l'échantillon (1) devant être testé et un passage de sortie (24) servant à dégager l'échantillon testé (1) et une porte de mesure (30) étant située à l'intérieur de la cuvette d'écoulement (20). Les éléments caractéristiques de l'invention consistent en ce que le corps de base (10) est complété d'un canal sous pression (12) approprié pour l'introduction du milieu de répartition (40) et d'un canal d'évacuation (13) approprié pour l'évacuation du milieu de répartition (40) ; et, en outre, il y a une ouverture de répartition (22) et une ouverture de sortie (23) dans la section de la cuvette d'écoulement (20) entre le passage d'entrée (21) et le passage de sortie (24) ; le canal sous pression (12) approprié pour l'introduction du milieu de répartition (40) est raccordé à l'ouverture de répartition (22) de la cuvette d'écoulement (20), alors que le canal d'évacuation (13) est raccordé à l'ouverture de sortie (23) ; l'ouverture de répartition (22) et l'ouverture de sortie (23) sont disposées sur la même surface latérale (11c) de l'enveloppe délimitante (11) ; l'axe principal (12a) du canal sous pression (12) s'ouvrant dans l'ouverture de répartition (22) est à un angle aigu d'obliquité (α) par rapport à la direction principale d'écoulement (1a) de l'échantillon (1), vu de la direction du passage d'entrée (21) de la cuvette d'écoulement (20) ; et en outre l'axe principal (13a) du canal d'évacuation (13) s'ouvrant dans l'ouverture de sortie (23) et la direction principale d'écoulement (1a) de l'échantillon (1) sont à un angle de cône obtus (β) l'un par rapport à l'autre, vu du passage d'entrée (21) ; et la porte de mesure (30) est située dans la section de la cuvette d'écoulement (20) entre l'ouverture de sortie (23) et le passage de sortie (25) ; l'une des électrodes conductrices de courant (60) est placée dans la partie de la cuvette d'écoulement (20) entre la porte de mesure (30) et le passage de sortie (25), alors que l'autre électrode conductrice de courant (70) est placée dans le seau du canal d'évacuation (13) de la cuvette d'écoulement (20) après l'ouverture de sortie (23) ; et dans l'environnement de la porte de mesure (30), il y a un corps de mesure électrique (31) contenant un premier capteur de mesure (31a) et un second capteur de mesure (31b).
PCT/HU2014/000013 2013-02-05 2014-02-04 Unité de test pour la détermination des caractéristiques physiques d'échantillons contenant des composants liquides WO2014122491A1 (fr)

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HU1300072A HU230238B1 (hu) 2013-02-05 2013-02-05 Mérőegység folyadékot tartalmazó minták fizikai jellemzőinek meghatározásához
HUHUP1300072 2013-02-05

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US10816550B2 (en) 2012-10-15 2020-10-27 Nanocellect Biomedical, Inc. Systems, apparatus, and methods for sorting particles

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WO2010106426A1 (fr) * 2009-03-17 2010-09-23 Silicon Biosystems S.P.A. Procédé d'isolement de particules
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WO2011143075A2 (fr) 2010-05-08 2011-11-17 Veridex, Llc Procédé simple et abordable pour l'immunophénotypage utilisant une préparation d'échantillons sur puce microfluidique avec cytométrie en image
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US20120211373A1 (en) 2011-02-22 2012-08-23 The Regents Of The University Of Michigan Microfluidic system for measuring cell barrier function
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EP2523004A1 (fr) 2004-01-26 2012-11-14 President and Fellows of Harvard College Procédé pour déterminer un analyte et test immunologique

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Publication number Priority date Publication date Assignee Title
WO1999009042A2 (fr) * 1997-08-13 1999-02-25 Cepheid Microstructures permettant de manipuler des echantillons fluides
EP2523004A1 (fr) 2004-01-26 2012-11-14 President and Fellows of Harvard College Procédé pour déterminer un analyte et test immunologique
US20120264134A1 (en) * 2006-03-31 2012-10-18 Fluxion Biosciences Inc. Methods and apparatus for the manipulation of particle suspensions and testing thereof
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US20090269767A1 (en) * 2008-04-10 2009-10-29 C/O Valtion Teknillinen Tutkimuskeskus Microfluidic chip devices and their use
WO2010106426A1 (fr) * 2009-03-17 2010-09-23 Silicon Biosystems S.P.A. Procédé d'isolement de particules
US20110192219A1 (en) * 2010-02-08 2011-08-11 Horiba, Ltd. Liquid sample analyzer
WO2011143075A2 (fr) 2010-05-08 2011-11-17 Veridex, Llc Procédé simple et abordable pour l'immunophénotypage utilisant une préparation d'échantillons sur puce microfluidique avec cytométrie en image
EP2490005A1 (fr) * 2011-02-18 2012-08-22 Koninklijke Philips Electronics N.V. Réseau de résistance microfluidique et dispositif microfluidique
US20120211373A1 (en) 2011-02-22 2012-08-23 The Regents Of The University Of Michigan Microfluidic system for measuring cell barrier function

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10816550B2 (en) 2012-10-15 2020-10-27 Nanocellect Biomedical, Inc. Systems, apparatus, and methods for sorting particles

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HUP1300072A2 (en) 2014-08-28

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