JP5313583B2 - Cell characteristic measuring device - Google Patents

Description

  The present invention relates to a cell characteristic measuring apparatus that includes a plurality of surface acoustic wave elements in which a propagation path in which cells are loaded is formed between input and output electrodes, and obtains physical characteristics of the cells.

  In general, a surface acoustic wave element includes a piezoelectric substrate, and an input electrode and an output electrode composed of comb-like electrode fingers provided on the piezoelectric substrate. In the surface acoustic wave element, when an electric signal is input to the input electrode, an electric field is generated between the electrode fingers, and the surface acoustic wave is excited by the piezoelectric effect and propagates on the piezoelectric substrate. Among these surface acoustic waves, detection of various substances and physical property values using surface acoustic wave elements that use a shear surface acoustic wave (SH-SAW) that is displaced in a direction perpendicular to the propagation direction An elastic wave sensor for performing measurement has been studied (Patent Document 1).

  In the acoustic wave sensor, there is a difference in the characteristics of the output signal output from the output electrode when the area of the object to be measured loaded on the piezoelectric substrate is electrically open and when it is short-circuited. By utilizing this, the dielectric constant and conductivity can be obtained as physical characteristics of the object to be measured. Further, when a concavo-convex structure is formed on the propagation path between the input electrode and the output electrode of the surface acoustic wave element and the object to be measured is loaded in the concave portion, the loaded object to be measured forms a pseudo film. This film is excited together with the piezoelectric substrate, and the density of the object to be measured can be obtained using the mass load effect in which the resonance frequency changes based on the mass of the film (Patent Document 2).

  The elastic wave sensor can also measure the physical characteristics of cells, and a cell physical property measuring device is known as a device for measuring changes in cell volume (Patent Document 3).

  In the cell physical property measuring apparatus 300 shown in FIG. 5, when an electric signal is input to the input electrode 304 of the surface acoustic wave element 302, an electric field is generated between the electrode fingers, and the surface acoustic wave is excited by the piezoelectric effect, A cell 308a, 308b, 308c, which is a measurement object, propagates on the propagation path 310 loaded on the 306, and is received by the output electrode 312 of the surface acoustic wave element. Based on the mass change amount of the surface acoustic wave detected from the signal received by the output electrode 312, the volume is measured as a physical characteristic of the cultured cells.

Japanese Patent No. 3481298 Japanese Patent No. 3248683 JP 2006-275798 A

  However, since the cells loaded on the cell physical property measuring apparatus 300 are randomly fixed on the propagation path 310, when the phase difference is detected based on the change in the propagation speed of the surface acoustic wave, the propagation is performed. When the width direction of the path 310 is considerably longer than the size of the cell 308, the detected phase change amount may be inaccurate, and a measurement error may occur. That is, the propagation path 310 is loaded with three cells 308a, 308b, and 308c, but the surface acoustic wave that propagates through the cells 308a and 308b and the surface acoustic wave that propagates through the cell 308c are output electrodes 312. Therefore, the phase change amount actually detected becomes smaller than the phase change amount originally detected corresponding to the three cells (308a, 308b, 308c). May end up. As described above, when a plurality of cells 308 are loaded in the width direction of the propagation path 310, the physical characteristics of the cells 308 may not be accurately measured.

  The present invention has been made in consideration of the above problems, and even when cells are sparsely loaded in the propagation path, it is possible to accurately measure the physical characteristics of the cells. It aims at providing a cell characteristic measuring device.

The cell characteristic measuring apparatus according to the present invention includes a plurality of surface acoustic wave elements that are formed so that a short-circuit propagation path in which a medium loaded with cells is arranged between input and output electrodes is formed , and arranged in parallel with each other. And the width of the short-circuit propagation path is such that the cell does not enter two or more in the width direction, the same signal is input from each input electrode, and based on the output signal output from each output electrode It is characterized in that physical characteristics of cells arranged in the plurality of surface acoustic wave elements are obtained.

  As physical characteristics of each cell, the mass change amount of each cell and the viscoelastic change amount of the medium can be obtained.

In addition, another cell characteristic measuring apparatus according to the present invention is a first surface acoustic wave device in which a first propagation path is formed as a short-circuit propagation path in which a first medium loaded with cells is placed between input and output electrodes. And a second surface acoustic wave element in which a second culture medium in which cells are loaded between the input and output electrodes is arranged , and a second propagation path which is an open propagation path having an amplitude / phase characteristic different from that of the first propagation path is formed. A plurality of SAW sensors having a width such that two or more cells do not enter in the width direction, and the SAW sensors are arranged in parallel, The same signal is input to the input electrode of the first surface acoustic wave element and the input electrode of the second surface acoustic wave element of each SAW sensor, the output signal from the output electrode of the first surface acoustic wave element, and the first Of the second surface acoustic wave element in the same SAW sensor as the one surface acoustic wave element And obtaining the physical properties of the respective cells based on the output signal from the force electrode.

  As the physical characteristics of each cell, at least one of the relative permittivity and conductivity of each cell can be determined, and at least one of the relative permittivity and conductivity of each medium can be determined.

  According to the present invention, it is possible to accurately measure the amount of change in cell mass and the amount of change in viscoelasticity of the culture medium as the physical characteristics of the cells by using a plurality of surface acoustic wave elements to form multichannels. In addition, by making a multichannel with a plurality of SAW sensors, it is possible to accurately measure the relative dielectric constant and conductivity of cells and culture media as the physical characteristics of the cells.

  Hereinafter, a first embodiment of the present invention will be described with reference to the drawings. FIG. 1 is an explanatory diagram of a configuration of a cell property measuring apparatus 10 according to an embodiment of the present invention, and FIG. 2 is an end view taken along the line II-II in FIG. The cell characteristic measuring apparatus 10 includes a SAW sensor 14, an oscillator 50 that generates a high-frequency electric signal, a distributor 52 that distributes the electric signal from the oscillator 50, an electric signal distributed from the distributor 52, and an elastic surface. An elastic wave detector 54 that measures an amplitude ratio, a phase difference, and the like with an output signal corresponding to a wave, and a physical characteristic calculator 56 that calculates a mass change amount and a viscoelastic change amount as physical characteristics of the cell. .

  The SAW sensor 14 includes surface acoustic wave elements 11, 12, and 13. The surface acoustic wave element 11 includes an input electrode 21 and an output electrode 31, a short-circuit propagation path 41 is formed between the input electrode 21 and the output electrode 31, and the surface acoustic wave element 12 includes the input electrode 22 and the output electrode. A short-circuit propagation path 42 is formed between the input electrode 22 and the output electrode 32, and the surface acoustic wave element 13 includes an input electrode 23 and an output electrode 33, and the input electrode 23 and the output electrode 33 are provided. A short-circuit propagation path 43 is formed between the two. The surface acoustic wave elements 11, 12, and 13 are disposed on the piezoelectric substrate 38 so as to be parallel to each other.

  The input electrodes 21, 22, and 23 are comb-shaped electrodes for exciting surface acoustic waves based on an electric signal input from the oscillator 50 via the distributor 52. The output electrodes 31, 32, and 33 are comb-shaped electrodes for receiving surface acoustic waves that are excited and propagated from the input electrodes 21, 22, and 23.

The short-circuit propagation paths 41, 42, and 43 are short-circuit propagation paths that are formed of the metal film 40 deposited on the piezoelectric substrate 38 and are electrically short-circuited. The length of the short-circuit propagation paths 41, 42, 43 in the width direction (arrow Y in FIG. 1) is slightly longer than the size according to the type of the cell 44 that is the object to be measured. It is preferable that the length does not enter two or more. The material of the metal film 40 is not particularly limited, but it is preferably formed of gold that is chemically stable for the cells 44. The piezoelectric substrate 38 is not particularly limited as long as it can propagate a sliding surface acoustic wave, but is preferably a 36-degree Y-plate X-propagating LiTaO ₃ .

  A medium 46 is placed on each surface of the short-circuit propagation paths 41, 42, and 43, and cells 44 are loaded on each medium 46 (see FIG. 2). The medium 46 is generally a solid medium, a liquid medium, a two-phase medium (for example, a gel medium) in which a solid medium and a liquid medium are mixed, and depends on the type of the cell 44 to be loaded. Identified. Among these, when a fixed medium is used as the medium 46, it is preferable to form a thin film having a thickness equal to or less than a half wavelength of the surface acoustic wave in order to suppress loss.

  The measurement of the physical property of the cell 44 by the cell property measuring apparatus 10 is performed as follows.

  First, the culture medium 46 is placed on each of the short-circuit propagation paths 41, 42, and 43, and the cells 44 that are objects to be measured are loaded on each culture medium 46. Thereafter, the electric signal from the oscillator 50 is distributed by the distributor 52, and the same signal is input to each of the elastic wave detector 54 and the input electrodes 21 (22, 23). In the input electrode 21, a surface acoustic wave is excited based on the input signal, propagates on the short-circuit propagation path 41 (X direction in FIG. 1), and is received by the output electrode 31. Similarly to the input electrode 21, the surface acoustic waves are excited at the input electrodes 22 and 23 based on the input signal, propagate through the short-circuit propagation path 42 (43), and are received at the output electrode 32 (33). .

  The elastic wave detector 54 detects the amplitude ratio, phase difference, and propagation delay difference between the signal output from the oscillator 50 and distributed by the distributor 52, and the output signals from the output electrodes 31, 32, 33. Is output to the target characteristic calculation unit 56.

When the purpose of loading the cells 44 on the culture medium 46 is to fix the cells, the physical property calculation unit 56 changes the mass of the cells 44 fixed to the culture media 46 of the short-circuit propagation paths 41, 42, 43. A quantity is calculated for each short-circuit propagation path. Since one cell 44 is fixed to the medium 46 of the short-circuit propagation path 41, two cells 44 are fixed to the medium 46 of the short-circuit propagation path 42 , and one cell 44 is fixed to the medium 46 of the short-circuit propagation path 43 . The amount of mass change corresponding to each number is calculated. In addition, the calculated value obtained by summing the respective mass change amounts is the total mass change amount of the cells 44 fixed to the short-circuit propagation paths 41, 42, 43. Further, in the physical property calculation unit 56, by preparing a correlation equation or correlation table between the mass change amount and the mass for each type of the cell 44 in advance, the medium is calculated from the calculated mass change amount of the cell 44. The mass of the cells 44 immobilized on 46 can be calculated. Furthermore, when the unit mass of the cell 44 is known, the number of the cells 44 can be calculated based on the calculated mass change amount of the cell 44 and fixed to the short-circuit propagation paths 41, 42, 43. It is also possible to calculate the total number of converted cells 44.

  When the purpose of loading the cells 44 on the medium 46 is cell growth, the physical property calculation unit 56 calculates the amount of viscoelasticity change including the cells 44 and the medium 46. This calculated change in viscoelasticity is also a calculated value corresponding to the number of cells 44 after growth for each medium 46 in the short-circuit propagation paths 41, 42, 43. In the physical property calculation unit 56, by preparing a correlation equation or correlation table between the viscoelastic change amount and the mass for each type of cell 44 in advance, the medium is calculated from the calculated viscoelastic change amount of the cell 44. The mass of the cell 44 loaded on 46 can be calculated. Furthermore, the total number of cells 44 grown in the short-circuit propagation paths 41, 42, and 43 can be calculated in the same manner as when the cells are immobilized.

  As described above, the cell characteristic measuring apparatus 10 includes the short-circuit propagation path 41 (42) in which the medium 46 loaded with the cells 44 is disposed between the input electrodes 21 (22, 23) and the output electrodes 31 (32, 33). , 43) formed on the basis of the output signals output from the output electrodes 31 (32, 33) by inputting signals from the input electrodes 21 (22, 23). The physical properties of the cell 44 are determined. In the cell characteristic measuring apparatus 10, the cell 44 is made multichannel by the plurality of surface acoustic wave elements 11, 12, and 13 in which the width of the short-circuit propagation paths 41, 42, 43 is determined in consideration of the size of the cell 44. As the physical characteristics, the mass change amount of the cells 44 and the viscoelastic change amount of the culture medium 46 can be accurately measured.

  Next, a second embodiment of the present invention will be described. FIG. 3 is an explanatory diagram of a configuration of the cell property measuring apparatus 10A according to the second embodiment. FIG. 4A is an end view of IVA-IVA in FIG. 3, and an end view of IVB-IVB in FIG. The same components as those in the first embodiment are denoted by the same reference numerals, and detailed description thereof is omitted.

  As shown in FIG. 3, the cell characteristic measuring apparatus 10A calculates SAW sensors 115, 116, 117, an oscillator 50, a distributor 52, an elastic wave detector 54, and a relative dielectric constant and conductivity as physical characteristics of the cell. And a physical characteristic calculation unit 56.

  The SAW sensor 115 includes a first surface acoustic wave element 111 and a second surface acoustic wave element 211, and the first surface acoustic wave element 111 includes an input electrode 121 and an output electrode 131, and the input electrode 121 and the output electrode 131 is formed with a short-circuit propagation path (first propagation path) 141, the second surface acoustic wave element 211 has an input electrode 221 and an output electrode 231, and the input electrode 221 and the output electrode 231 An open propagation path (second propagation path) 241 is formed between them. Similarly, the SAW sensor 116 includes a first surface acoustic wave element 112 and a second surface acoustic wave element 212, and the first surface acoustic wave element 112 includes an input electrode 122 and an output electrode 132. A short-circuit propagation path 142 is formed between the input electrode 222 and the output electrode 232, and the second surface acoustic wave element 212 includes an input electrode 222 and an output electrode 232. , An open propagation path 242 is formed. The SAW sensor 117 includes a first surface acoustic wave element 113 and a second surface acoustic wave element 213, and the first surface acoustic wave element 113 includes an input electrode 123 and an output electrode 133. A short-circuit propagation path 143 is formed between the output electrode 133 and the second surface acoustic wave element 213 includes an input electrode 223 and an output electrode 233, and between the input electrode 223 and the output electrode 233, An open propagation path 243 is formed. Further, the SAW sensors 115, 116, and 117 are disposed on the piezoelectric substrate 138 so as to be parallel to each other.

  In the open propagation paths 241, 242, and 243, a part of the metal film is peeled off, and the metal film 140 is formed so that the piezoelectric substrate 138 is exposed. Therefore, the open region 144 where the piezoelectric substrate 138 is exposed is electrically open. Moreover, it is preferable that the length of the open propagation paths 241 242 243 in the width direction (arrow Y in FIG. 3) is slightly longer than the size according to the type of the cell 44 that is the object to be measured.

  The input electrodes 121, 221, 122, 222, 123, and 223 are formed in the same manner as the input electrode 21 of the cell characteristic measurement apparatus 10, and the output electrodes 131, 231, 132, 232, 133, and 233 are cell characteristic measurements It is formed in the same manner as the output electrode 31 of the device 10. The piezoelectric substrate 138 is formed in the same manner as the piezoelectric substrate 38, and the metal film 140 is formed in the same manner as the metal film 40.

  The measurement of the physical property of the cell 44 by the cell property measuring apparatus 10A is performed as follows.

  First, the culture medium 46 is placed on each of the short-circuit propagation paths 141, 142, and 143 and the open propagation paths 241, 242, and 243, and the cell 44 that is the object to be measured is loaded on each culture medium 46. Thereafter, the electrical signal from the oscillator 50 is distributed by the distributor 52 and the same signal is input to each of the input electrodes 121, 221, 122, 222, 123, and 223.

  A surface acoustic wave is excited on the input electrode 121 of the SAW sensor 115 based on the input signal, propagates on the short-circuit propagation path 141 and is received by the output electrode 131, and the input signal is converted to the input signal 221. Based on this, the surface acoustic wave is excited, propagates on the open propagation path 241, and is received by the output electrode 231. Both output signals extracted from the surface acoustic waves received by the output electrodes 131 and 231 are compared by the elastic wave detector 54 to detect an amplitude ratio and a phase difference. Similarly, in each of the SAW sensors 116 (117), a surface acoustic wave is excited based on the input signal and propagates through the short-circuit propagation path 142 (143) and the open propagation path 242 (243) to be output. Both output signals received by the electrodes 132 and 232 and extracted from the surface acoustic wave are compared by the acoustic wave detector 54, and received by the output electrode 132 (133) and the output electrode 232 (233) and extracted from the surface acoustic wave. Both output signals are compared by the elastic wave detector 54, and the amplitude ratio and phase difference of each are detected.

  The amplitude ratio and phase difference in the SAW sensors 115, 116, and 117 detected by the elastic wave detector 54 are output to the relative dielectric constant / conductivity calculator 58. When the purpose of loading the cells 44 on the medium 46 is to fix the cells, the relative permittivity / conductivity calculator 58 determines the relative permittivity and conductivity of the medium 46 for each of the SAW sensors 115, 116, and 117. Calculated. When the purpose of loading the cells 44 on the medium 46 is cell proliferation, the relative permittivity / conductivity calculating unit 58 integrates the cells 44 and the medium 46 for each SAW sensor 115, 116, 117. The calculated relative dielectric constant and conductivity are calculated. The calculated relative permittivity and conductivity correspond to the number of cells 44 grown and changes in the medium 46.

  As described above, the cell characteristic measuring apparatus 10A is a short circuit in which the medium (first medium) 46 loaded with the cells 44 is disposed between the input electrode 121 (122, 123) and the output electrode 131 (132, 133). A cell 44 is loaded between the first surface acoustic wave element 111 (112, 113) in which the propagation path 141 (142, 143) is formed, the input electrode 221 (222, 223), and the output electrode 231 (232, 233). And a SAW sensor 115 (116, 117) in which an open propagation path 241 (242, 243) having an amplitude / phase characteristic different from that of the short-circuit propagation path 141 (142, 143) is formed. The SAW sensors 115, 116, 117 are arranged in parallel, and input the same signal to the input electrodes 121 (122, 123) and the output electrodes 131 (132, 133). Output signals from the output electrodes 131 (132, 133) of the first surface acoustic wave element 111 (112, 113) and outputs from the output electrodes 231 (232, 233) of the second surface acoustic wave element 211 (212, 213) Based on the signal, the physical characteristics of the cell 44 are determined.

  In the cell characteristic measuring apparatus 10A, the width of the open propagation paths 241, 242, and 243 is multi-channeled by the SAW sensors 115, 116, and 117 determined in consideration of the size of the cell 44, whereby the physical characteristics of the cell 44 are obtained. As described above, the relative dielectric constant and conductivity of the cell 44 and the culture medium 46 can be accurately measured.

  In the cell characteristic measuring apparatus 10, the number of surface acoustic wave elements is such that the surface acoustic wave elements 11, 12, 13 and three surface acoustic wave elements are arranged in parallel. Is not limited. In the cell characteristic measuring apparatus 10A, the SAW sensors 115, 116, and 117 and the three SAW sensors are arranged in parallel, but the number is not limited as long as the number is two or more.

  In addition, the cells 44 loaded on the short-circuit propagation paths 41, 42, 43, 141, 142, 143, and the open propagation paths 241, 242, 243 are not limited to the same type, but are different for each propagation path. 44 may be loaded.

  It should be noted that the present invention is not limited to the above-described embodiment, and various configurations can be adopted without departing from the gist of the present invention.

It is explanatory drawing of a structure of the cell characteristic measuring apparatus which concerns on 1st Embodiment of this invention. It is the II-II end view of FIG. It is explanatory drawing of a structure of the cell characteristic measuring apparatus which concerns on 2nd Embodiment of this invention. 4A is an end view of IVA-IVA in FIG. 3, and FIG. 4B is an end view of IVB-IVB in FIG. 3. It is explanatory drawing of the conventional cell characteristic measuring apparatus.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10, 300 ... Cell physical property measuring apparatus 11-13 ... SAW element 14, 115-117 ... SAW sensors 21-23, 121-123, 221-223, 304 ... Input electrodes 31-33, 131-133, 231- 233, 312 ... output electrodes 38, 138, 306 ... piezoelectric substrate 40 ... metal films 41-43, 141-143 ... short-circuit propagation paths 44, 308a-308c ... cells 46 ... medium 50 ... oscillator 52 ... distributor 54 ... elastic wave Detector 56 ... Physical characteristic calculator 58 ... Relative permittivity / conductivity calculator 111 to 113 ... first surface acoustic wave element 140 ... metal film 144 ... open region 211 to 213 ... second surface acoustic wave element 241 to 243 ... Open propagation path 302 ... Surface acoustic wave element 310 ... Propagation path

Claims (6)

A short-circuit propagation path in which a medium loaded with cells is arranged between the input and output electrodes is formed , and includes a plurality of surface acoustic wave elements arranged in parallel with each other ,
The width of the short-circuit propagation path is such a length that two or more cells do not enter in the width direction,
Cell characteristics characterized in that the same signal is input from each of the input electrodes, and physical characteristics of cells arranged in the plurality of surface acoustic wave elements are obtained based on output signals output from the output electrodes. measuring device. The device for measuring cell characteristics according to claim 1,
An apparatus for measuring cell characteristics, wherein the amount of mass change of each cell is determined as a physical characteristic of each cell. In the cell characteristic measuring device according to claim 1 or 2,
A cell property measuring apparatus, wherein the amount of change in viscoelasticity of each medium is determined as a physical property of each cell. A first surface acoustic wave device having a first propagation path as a short-circuit propagation path in which a first medium loaded with cells between the input and output electrodes is disposed, and a second surface loaded with cells between the input and output electrodes A plurality of SAW sensors having a second surface acoustic wave element in which a culture medium is disposed and a second propagation path that is an open propagation path having different amplitude and phase characteristics from the first propagation path is formed;
The widths of the first propagation path and the second propagation path are such that the cell does not enter two or more in the width direction,
The SAW sensors are arranged in parallel,
The same signal is input to the input electrode of the first surface acoustic wave element and the input electrode of the second surface acoustic wave element of each SAW sensor, the output signal from the output electrode of the first surface acoustic wave element, A cell characteristic measuring apparatus characterized in that a physical characteristic of each cell is obtained based on an output signal from an output electrode of a second surface acoustic wave element in the same SAW sensor as the first surface acoustic wave element. The device for measuring cell characteristics according to claim 1,
At least one of the relative dielectric constant or conductivity of each cell is obtained as the physical property of each cell. In the cell characteristic measuring device according to claim 4 or 5,
As a physical characteristic of each cell, at least one of a relative permittivity and a conductivity of each medium is obtained.

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