JP2014224741A - Magnetic particulate detecting device and magnetic particulate detecting method - Google Patents
Magnetic particulate detecting device and magnetic particulate detecting method Download PDFInfo
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Abstract
Description
本発明は、磁性微粒子検出装置及び磁性微粒子検出方法に関する。 The present invention relates to a magnetic particle detection apparatus and a magnetic particle detection method.
微粒子材料は、バイオテクノロジーにおいて幅広い利用が期待されている。なかでもナノ粒子は細胞・DNA・タンパク質などの分離や各種アッセイ診断やドラッグデリバリーシステム(DDS)など幅広い領域での利用が期待されている。特に近年バイオテクノロジーや医療にナノテクノロジーを応用したバイオテクノロジーの展開に対する期待が高まり、研究開発が活発化されている。 Fine particle materials are expected to be widely used in biotechnology. Among these, nanoparticles are expected to be used in a wide range of fields such as separation of cells, DNA, proteins, etc., various assay diagnosis, and drug delivery system (DDS). In particular, in recent years, expectations for the development of biotechnology that applies nanotechnology to biotechnology and medicine have increased, and research and development have been activated.
ナノ粒子の特徴として、粒径をナノサイズにすることにより単位体積当たりの表面積(吸着や反応の場として利用できる)が著しく増大する点が挙げられる。したがって、ナノ粒子を診断薬へ応用する場合、より短時間に高感度で診断が可能なシステムの構築が可能である。ナノ粒子材料に薬剤を包み込みDDSに応用した場合、通常の薬剤では簡単に到達できない患部に薬剤を送り込むことも可能になると期待されている。 A feature of the nanoparticles is that the surface area per unit volume (which can be used as a field for adsorption or reaction) is remarkably increased by making the particle size nano size. Therefore, when applying nanoparticles to a diagnostic agent, it is possible to construct a system capable of performing diagnosis with higher sensitivity in a shorter time. When a drug is encapsulated in a nanoparticle material and applied to DDS, it is expected that the drug can be delivered to an affected area that cannot be easily reached by a normal drug.
イミュノアッセイではその磁性微粒子の量を計測することが必要になり、これまでに数10kHzの周波数の正弦波交流を印加してその3倍波(第3高調波)成分を検出する方法が一般的に知られている。
図7は従来の磁性微粒子検出方法の原理の説明図である。
従来の磁性微粒子検出方法では、図7(a)に示すように磁性微粒子に微弱な正弦波交流磁場(破線)を印加すると、その微粒子のM−H特性の直線部で変調されるので、変調信号は図7(b)の破線のように正弦波が出力される。ここで、M−H特性が飽和する2つの変曲点(Mk,−Mk)を越える大きな正弦波交流磁場(実線)を印加すると、両変曲点がリミッタの作用をして波形が歪み、変調信号は図7(b)の実線のように正弦波の頭がカットされた方形波に近い信号となる。この方形波は式(1)のように奇数高調波成分のみを含むので、基本波(印加交流磁場の周波数)の影響を受けることはなく、
In the immunoassay, it is necessary to measure the amount of the magnetic fine particles, and a method of detecting the third harmonic (third harmonic) component by applying a sinusoidal alternating current with a frequency of several tens of kHz is generally used so far. Known.
FIG. 7 is an explanatory diagram of the principle of a conventional magnetic fine particle detection method.
In the conventional magnetic fine particle detection method, as shown in FIG. 7A, when a weak sinusoidal alternating magnetic field (broken line) is applied to the magnetic fine particles, the magnetic fine particles are modulated at the linear portion of the MH characteristic. As a signal, a sine wave is output as shown by a broken line in FIG. Here, when a large sine wave AC magnetic field (solid line) exceeding two inflection points (Mk, -Mk) at which the MH characteristic is saturated, both inflection points act as a limiter, and the waveform is distorted. The modulation signal is a signal close to a square wave with the sine wave head cut off as shown by the solid line in FIG. Since this square wave contains only odd harmonic components as in equation (1), it is not affected by the fundamental wave (frequency of the applied AC magnetic field)
最も大きな第3高調波成分を検出することで磁性微粒子の量を計測することができる。なお、FFP(Field−Free Pont)に、Hac=2Hk sin(ω0 t)の励起磁場を印加した場合、第三次高調波の振幅は、(1/3)×(4A/3π)と表すことができる。 The amount of magnetic fine particles can be measured by detecting the largest third harmonic component. When an excitation magnetic field of H ac = 2H k sin (ω 0 t) is applied to FFP (Field-Free Pont), the amplitude of the third harmonic is (1/3) × (4A / 3π) It can be expressed as.
しかしながら、上記した従来の磁性微粒子検出方法では、変曲点以上の大きさの正弦波磁場を印加することが必須となり、そのためには大きな交流電源とコイルが必要となる。
本発明は、上記状況に鑑みて、印加交流磁場の大小にかかわらず、大きな第2高調波成分を検出することにより、微粒子の量を高感度で計測することができる磁性微粒子検出装置及び磁性微粒子検出方法を提供することを目的とする。
However, in the above-described conventional magnetic fine particle detection method, it is essential to apply a sinusoidal magnetic field having a magnitude greater than the inflection point, which requires a large AC power source and coil.
In view of the above situation, the present invention provides a magnetic fine particle detection device and a magnetic fine particle that can measure the amount of fine particles with high sensitivity by detecting a large second harmonic component regardless of the magnitude of the applied AC magnetic field. An object is to provide a detection method.
本発明は、上記目的を達成するために、
〔1〕磁性微粒子検出装置において、差動検出コイルと、交流磁場印加コイルと、直流磁場付与装置とが同軸状に重なって配置されており、かつ前記差動検出コイルの出力はセンサーに接続されており、前記交流磁場印加コイルに正弦波交流を印加すると共に前記前記直流磁場付与装置に直流磁場を加えて変曲点まで動作点を移動して第2高調波を計測するように構成したことを特徴とする。
In order to achieve the above object, the present invention provides
[1] In the magnetic fine particle detection device, the differential detection coil, the AC magnetic field applying coil, and the DC magnetic field applying device are coaxially overlapped and the output of the differential detection coil is connected to the sensor. And applying a sine wave alternating current to the alternating magnetic field applying coil, applying a direct magnetic field to the direct current magnetic field applying device, moving the operating point to an inflection point, and measuring the second harmonic. It is characterized by.
〔2〕上記〔1〕記載の磁性微粒子検出装置において、前記直流磁場付与装置が直流磁場印加コイルであることを特徴とする。
〔3〕上記〔1〕記載の磁性微粒子検出装置において、前記直流磁場付与装置が永久磁石であることを特徴とする。
〔4〕上記〔1〕記載の磁性微粒子検出装置において、前記センサーがスペクトラムアナライザであることを特徴とする。
[2] The magnetic fine particle detection device according to [1], wherein the DC magnetic field applying device is a DC magnetic field applying coil.
[3] The magnetic fine particle detection device according to [1], wherein the DC magnetic field applying device is a permanent magnet.
[4] The magnetic fine particle detection device according to [1], wherein the sensor is a spectrum analyzer.
〔5〕上記〔1〕記載の磁性微粒子検出装置において、前記センサーがSQUID磁気検出装置であり、前記差動検出コイルに流れる電流を磁気的に計測することを特徴とする。
〔6〕上記〔1〕記載の磁性微粒子検出装置において、前記センサーがフラックスゲートセンサーであり、前記差動検出コイルに流れる電流を磁気的に計測することを特徴とする。
[5] The magnetic fine particle detection device according to [1], wherein the sensor is a SQUID magnetic detection device, and the current flowing through the differential detection coil is magnetically measured.
[6] The magnetic fine particle detection device according to [1], wherein the sensor is a fluxgate sensor, and a current flowing through the differential detection coil is magnetically measured.
〔7〕上記〔1〕記載の磁性微粒子検出装置において、前記センサーがMI効果素子であり、前記差動検出コイルに流れる電流を磁気的に計測することを特徴とする。
〔8〕磁性微粒子検出方法において、差動検出コイルと、交流磁場印加コイルと、直流磁場付与装置とが同軸状に重なるように配置し、前記差動検出コイルの出力はセンサーに接続し、前記交流磁場印加コイルに正弦波交流を印加すると共に前記直流磁場付与装置に直流磁場を加えて変曲点まで動作点を移動して第2高調波を計測することを特徴とする。
[7] The magnetic fine particle detection device according to [1], wherein the sensor is an MI effect element and magnetically measures a current flowing through the differential detection coil.
[8] In the magnetic particle detection method, the differential detection coil, the AC magnetic field application coil, and the DC magnetic field applying device are arranged so as to be coaxially overlapped, and an output of the differential detection coil is connected to a sensor, A second harmonic is measured by applying a sine wave alternating current to the alternating magnetic field application coil and applying a direct magnetic field to the direct magnetic field applying device to move the operating point to the inflection point.
〔9〕上記〔8〕記載の磁性微粒子検出方法において、前記半波整流波形が [9] In the magnetic particle detection method according to [8], the half-wave rectified waveform is
であることを特徴とする。 It is characterized by being.
本発明によれば、印加交流磁場の大小にかかわらず、大きな第2高調波成分を検出することにより微粒子の量を高感度で計測することができる。
また、正弦波交流と共に直流磁場を加えて変曲点まで動作点を移動して計測することにより、変曲点以上の大きさの正弦波磁場を印加する必要がなくなり、大きな交流電源やコイルを設ける必要はなくなり、磁性微粒子検出装置の小型化・コンパクト化を図ることができる。
According to the present invention, the amount of fine particles can be measured with high sensitivity by detecting a large second harmonic component regardless of the magnitude of the applied AC magnetic field.
In addition, by applying a DC magnetic field together with a sinusoidal alternating current and moving the operating point to the inflection point, there is no need to apply a sinusoidal magnetic field larger than the inflection point. There is no need to provide it, and the magnetic particle detector can be made smaller and more compact.
本発明の磁性微粒子検出装置は、差動検出コイルと、交流磁場印加コイルと、直流磁場付与装置とが同軸状に重なって配置されており、かつ前記差動検出コイルの出力はセンサーに接続されており、前記交流磁場印加コイルに正弦波交流を印加すると共に前記前記直流磁場付与装置に直流磁場を加えて変曲点まで動作点を移動して第2高調波を計測するように構成した。 In the magnetic fine particle detection device of the present invention, a differential detection coil, an AC magnetic field applying coil, and a DC magnetic field applying device are coaxially arranged, and the output of the differential detection coil is connected to a sensor. In addition, a sine wave AC is applied to the AC magnetic field applying coil, and a DC magnetic field is applied to the DC magnetic field applying device to move an operating point to an inflection point and measure the second harmonic.
以下、本発明の磁性微粒子検出方法について詳細に説明する。
図1は本発明の磁性微粒子検出方法の原理の説明図である。
本発明では、図1(a)に示すように、どちらかの変曲点において正弦波磁場を印加すると、図1(b)に示すように、その正弦波交流の大きさが小さいとき(破線)も、大きいとき(実線)も半波整流波形のような変調信号が得られる。この半波整流波形は数式(2)のように偶数高調波成分のみを含むので、
Hereinafter, the magnetic particle detection method of the present invention will be described in detail.
FIG. 1 is an explanatory diagram of the principle of the magnetic particle detection method of the present invention.
In the present invention, as shown in FIG. 1A, when a sinusoidal magnetic field is applied at either inflection point, the magnitude of the sinusoidal alternating current is small as shown in FIG. However, when it is large (solid line), a modulated signal like a half-wave rectified waveform can be obtained. Since this half-wave rectified waveform contains only even harmonic components as shown in Equation (2),
基本波(印加交流磁場の周波数)の影響を受けることはなく、かつ、印加磁場の大小にかかわらず、大きな第2高調波成分を検出することで微粒子の量を高感度で計測することができる。なお、KFP(Knee−Free Pont)にHac=2Hk sin(ω0 t)の励起磁場を印加した場合、第二次高調波の振幅は、4A/3πと表すことができる。 It is not affected by the fundamental wave (frequency of applied AC magnetic field), and the amount of fine particles can be measured with high sensitivity by detecting a large second harmonic component regardless of the magnitude of the applied magnetic field. . Note that when an excitation magnetic field of H ac = 2H k sin (ω 0 t) is applied to KFP (Knee-Free Pont), the amplitude of the second harmonic can be expressed as 4A / 3π.
図2は本発明の実施例(実験装置)を示す磁性微粒子検出装置の構成図である。
この図において、1は差動検出コイル、2は交流磁場印加コイル、3は直流磁場付与手段としての直流磁場印加コイルであり、これらの差動検出コイル1と、交流磁場印加コイル2と、直流磁場印加コイル3は同軸状に重なるように構成されており、差動検出コイル1の出力はスペクトラムアナライザ4に接続されている。
FIG. 2 is a configuration diagram of a magnetic fine particle detection apparatus showing an embodiment (experimental apparatus) of the present invention.
In this figure, 1 is a differential detection coil, 2 is an AC magnetic field application coil, 3 is a DC magnetic field application coil as a DC magnetic field applying means, and these
なお、ここで、作図上、差動検出コイル1と交流磁場印加コイル2と直流磁場印加コイル3は展開して描かれているが、差動検出コイル1と交流磁場印加コイル2と直流磁場印加コイル3は同軸状に重なって配置されるように構成する。
また、図2に示した磁性微粒子検出装置に代えて、直流磁場印加コイルは永久磁石とし、スペクトラムアナライザ4は、SQUID磁気検出装置や、フラックスゲートセンサーやMI効果素子を用いるようにすることができる。
Here, in the drawing, the
Further, instead of the magnetic fine particle detection device shown in FIG. 2, the DC magnetic field application coil may be a permanent magnet, and the
図3は本発明の第1実施例を示す磁性微粒子(商品名リゾビスト)を用いた従来の磁性微粒子検出方法と本発明の磁性微粒子検出方法による結果を示す図である。
図4は本発明の第2実施例を示す磁性微粒子(商品名フェリデックス)を用いた従来の磁性微粒子検出方法と本発明の磁性微粒子検出方法による結果を示す図である。
磁性微粒子(商品名リゾビスト)5を差動検出コイル1の一方のコイル内にセットして、交流磁場印加コイル2に正弦波交流の振幅を変えて第三高調波を計測した結果(▲)および、交流磁場印加コイル2に正弦波交流と共に直流磁場印加コイル3に直流磁場を加えて変曲点まで動作点を移動して第2高調波を計測した結果(●)が図3に示されている。図4は第1実施例と同様の実験を、別の磁性微粒子(商品名フェリデックス)5′を用いて得られた結果が図4に示されている。
FIG. 3 is a diagram showing the results of a conventional magnetic fine particle detection method using magnetic fine particles (trade name Rhizovist) according to the first embodiment of the present invention and the magnetic fine particle detection method of the present invention.
FIG. 4 is a diagram showing the results of a conventional magnetic fine particle detection method using magnetic fine particles (trade name Feridex) according to the second embodiment of the present invention and the magnetic fine particle detection method of the present invention.
The result of measuring the third harmonic by setting the magnetic fine particles (trade name Resovist) 5 in one coil of the
何れも明らかに、正弦波と共に直流磁場を加えて変曲点まで動作点を移動して第2高調波を計測した方が大きな信号が得られており、この磁性微粒子検出方法の優位性が示されている。
以下、実験装置と実験条件について説明する。
実験装置は、図2の構成とし、実験条件としては、励起磁場周波数:20.02kHz、励起磁場強度:実験ごとに変化、直流磁場強度:実験ごとに変化、サンプル名は、品名:Resovist(商品名リゾビスト)、粒子径:57nm(超常磁性)、容量:70μLである。
Obviously, a larger signal was obtained when the second harmonic was measured by applying a DC magnetic field along with a sine wave and moving the operating point to the inflection point, indicating the superiority of this magnetic particle detection method. Has been.
Hereinafter, an experimental apparatus and experimental conditions will be described.
The experimental apparatus has the configuration shown in FIG. 2 and the experimental conditions are: excitation magnetic field frequency: 20.02 kHz, excitation magnetic field strength: change for each experiment, DC magnetic field strength: change for each experiment, sample name: product name: Resovist (product Nominal resovist), particle size: 57 nm (superparamagnetism), volume: 70 μL.
図5は直流磁場強度Bdcを変化させた場合を示す特性図である。
ここで、実験条件として、励起磁場強度:10.23mTp-p Bk =3.28mTであり、図5の横軸は直流磁場強度Bdc(mT)、縦軸は信号(mVrms /Hz1/2 )、●は第2高調波、▲は第3高調波を示している。
図6は交流磁場強度Bacを変化させた場合を示す特性図である。
FIG. 5 is a characteristic diagram showing a case where the DC magnetic field strength B dc is changed.
Here, as experimental conditions, the excitation magnetic field intensity is 10.23 mT pp B k = 3.28 mT, the horizontal axis of FIG. 5 is the DC magnetic field intensity B dc (mT), and the vertical axis is the signal (mV rms / Hz 1 / Hz). 2 ), ● indicates the second harmonic, and ▲ indicates the third harmonic.
FIG. 6 is a characteristic diagram showing a case where the alternating magnetic field strength Bac is changed.
この図の横軸は交流磁場強度Bac(mTp-p )、縦軸は信号(mVrms /Hz1/2 )、●は第2高調波(Bdc=3.28mT)(15mVrms /√Hz)、▲は第3高調波(Bdc=0mT)(10mVrms /√Hz)である。
誘導コイルでの検出のため、第3高調波を基準とした場合、第2高調波の信号は、15×1.5=22.5mVrms /√Hzとなり、約2.25倍大きい。もし、Bac=2Bk となるような励起磁場であれば(Bac=13.12mTp-p )、より理論的な値である3倍に近い信号が得られる。
In this figure, the horizontal axis is AC magnetic field intensity B ac (mT pp ), the vertical axis is signal (mV rms / Hz 1/2 ), and ● is the second harmonic (B dc = 3.28 mT) (15 mV rms / √Hz. ) And ▲ are third harmonics (B dc = 0 mT) (10 mV rms / √Hz).
When the third harmonic is used as a reference for detection by the induction coil, the signal of the second harmonic is 15 × 1.5 = 22.5 mV rms / √Hz, which is about 2.25 times larger. If the excitation magnetic field is such that B ac = 2B k (B ac = 13.12 mT pp ), a signal closer to three times the more theoretical value can be obtained.
次に、KFPを用いる利点について説明する。
(1)FFPを用いた場合、無限大に近い励起磁場Hacでなければ、完全な矩形波を出すことはできない。その結果、奇数高調波は、Hac=2Hk の場合、完全な矩形波に比べ、信号が33%に減少する。
(2)奇数高調波は、Hac>Hk の場合でなければ計測することができない。
(3)偶数高調波は、Hacの大きさによらず、完全な半波を出すことができるため、奇数高調波のように信号が減少しない。そのため、第2高調波の信号はHac=2Hk の場合、第3高調波よりも約3倍(≒1/0.33)大きくなる。
(4)偶数高調波は、Hac<Hk の場合でも計測することができる。
Next, advantages of using KFP will be described.
(1) When FFP is used, a complete rectangular wave cannot be produced unless the excitation magnetic field H ac is close to infinity. As a result, odd harmonics reduce the signal to 33% when H ac = 2H k compared to a perfect square wave.
(2) Odd harmonics cannot be measured unless H ac > H k .
(3) Since even harmonics can produce complete half waves regardless of the magnitude of H ac , the signal does not decrease unlike odd harmonics. Therefore, the second harmonic signal is approximately three times (≈1 / 0.33) larger than the third harmonic signal when H ac = 2H k .
(4) Even harmonics can be measured even when H ac <H k .
なお、上記した実施例では、微粒子の検出について示したが、図2と同じような構成でイメージングを行う方法があり、従来法では直流磁場の大きさを変えることでFFPの空間的位置を移動させて、第3高調波を検出してイメージングを行っている。これに対して、ここでは同様に直流磁場の大きさを変えることでKFPの空間的位置を移動させて、第2高調波を検出してイメージングを行うこともできる。この場合も微粒子の検出と同様に小さな交流磁場でもイメージングを行うことができる利点がある。 In the above-described embodiments, detection of fine particles has been described. However, there is a method of performing imaging with the same configuration as in FIG. 2, and in the conventional method, the spatial position of the FFP is moved by changing the magnitude of the DC magnetic field. Thus, imaging is performed by detecting the third harmonic. On the other hand, here, it is also possible to perform imaging by detecting the second harmonic by moving the spatial position of the KFP by changing the magnitude of the DC magnetic field. In this case as well, there is an advantage that imaging can be performed even with a small alternating magnetic field as in the case of detection of fine particles.
また、本発明は上記実施例に限定されるものではなく、本発明の趣旨に基づき種々の変形が可能であり、これらを本発明の範囲から排除するものではない。 Further, the present invention is not limited to the above-described embodiments, and various modifications can be made based on the spirit of the present invention, and these are not excluded from the scope of the present invention.
本発明の磁性微粒子検出装置及び磁性微粒子検出方法は、印加交流磁場の大小にかかわらず、大きな第2高調波成分を検出することにより、微粒子の量を高感度で計測する磁性微粒子検出装置及び磁性微粒子検出方法として利用することができる。 The magnetic fine particle detection apparatus and magnetic fine particle detection method according to the present invention detect a large second harmonic component regardless of the magnitude of the applied AC magnetic field, thereby measuring the amount of fine particles with high sensitivity and magnetic It can be used as a fine particle detection method.
1 差動検出コイル
2 交流磁場印加コイル
3 直流磁場印加コイル(直流磁場付与装置)
4 スペクトラムアナライザ
5,5′ 磁性微粒子
DESCRIPTION OF
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