JP2007319035A - Device for collecting physiologically active substance and method for the same - Google Patents

Device for collecting physiologically active substance and method for the same Download PDF

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JP2007319035A
JP2007319035A JP2006150352A JP2006150352A JP2007319035A JP 2007319035 A JP2007319035 A JP 2007319035A JP 2006150352 A JP2006150352 A JP 2006150352A JP 2006150352 A JP2006150352 A JP 2006150352A JP 2007319035 A JP2007319035 A JP 2007319035A
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electrode
active substance
physiologically active
liquid
dna
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Kiyohiko Tateyama
清彦 館山
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Olympus Corp
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<P>PROBLEM TO BE SOLVED: To make it possible to collect a physiologically active substance capable of being applied as it is to a cell manipulation such as gene introduction, etc., and also collect the physiologically active substance directly from a living body specimen such as cells, etc. <P>SOLUTION: This device 1 for collecting the physiologically active substance is equipped with a liquid-holding part 11 for holding the liquid L containing the cells holding the physiologically active substance in their inside or the liquid L containing the physiologically active substance, a first electrode 12 installed as making a contact with the liquid, a second electrode 13 installed as relatively transferrable to the liquid-holding part 11, a transferring means for relatively transferring the second electrode 13 between the position of making the contact with the cell or the liquid L and a leaving position from them, and an electric source 15 for impressing a pulse electric voltage biased as a reverse polarity to the electric charge of the physiologically active substance based on the electric potential of the first electrode 12 as a relative standard, toward the second electrode 13 inserted into the cell or liquid L. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

本発明は、生理活性物質採取装置および生理活性物質採取方法に関するものである。   The present invention relates to a physiologically active substance collection device and a physiologically active substance collection method.

従来、電極間に電圧をかけて、ＤＮＡ（DeoxyriboNucleic Acid）を採取する装置としてＤＮＡピンセットが知られている（例えば、非特許文献１参照。）。
このＤＮＡピンセットは、電極となるＤＮＡピンセットの先端をＤＮＡ溶液に接触させ、高周波電圧を加えることにより、両電極間にＤＮＡを集めるものであり、電極先端にＤＮＡが付着するためＤＮＡ溶液内からＤＮＡを採取することができる。
Gen Hashiguchi, et al., “DNAManipulation and Retrieval from an Aqueous Solution with MicromachinedNanotweezers” Analytical Chemistry, vol. 75, No. 17,September 1, 2003, pp.4347-4350 Conventionally, a DNA tweezer is known as a device that collects DNA (DeoxyriboNucleic Acid) by applying a voltage between electrodes (see, for example, Non-Patent Document 1).
This DNA tweezers collects DNA between both electrodes by bringing the tip of the DNA tweezers to be an electrode into contact with the DNA solution and applying a high-frequency voltage. Can be collected.
Gen Hashiguchi, et al., “DNAManipulation and Retrieval from an Aqueous Solution with MicromachinedNanotweezers” Analytical Chemistry, vol. 75, No. 17, September 1, 2003, pp.4347-4350

しかしながら、ＤＮＡピンセットは電極間においてＤＮＡを採取するため、採取されたＤＮＡを遺伝子導入などの細胞操作に適用することが困難であるという問題がある。すなわち、遺伝子導入等の細胞操作を行うには、微細な針部にＤＮＡを保持させる必要があるので、ＤＮＡピンセットの先端の電極間に挟まれるようにして採取されたＤＮＡは、そのまま細胞に導入することはできない。
さらに、ＤＮＡピンセットは、ＤＮＡ溶液内のＤＮＡを採取することはできるものの、細胞のような生体試料内から直接採取することができないという不都合がある。 However, since DNA tweezers collect DNA between electrodes, there is a problem that it is difficult to apply the collected DNA to cell operations such as gene transfer. That is, in order to perform cell operations such as gene transfer, it is necessary to hold the DNA in a fine needle, so that the DNA collected so as to be sandwiched between the electrodes at the tip of the DNA tweezers is directly introduced into the cell. I can't do it.
Furthermore, although DNA tweezers can collect DNA in a DNA solution, there is a disadvantage that it cannot be directly collected from within a biological sample such as a cell.

本発明は上述した事情に鑑みてなされたものであって、遺伝子導入等の細胞操作にそのまま適用可能な状態で生理活性物質を採取でき、細胞等の生体試料からも直接的に生理活性物質を採取することができる生理活性物質採取装置および生理活性物質採取方法を提供することを目的としている。   The present invention has been made in view of the above-described circumstances, and a physiologically active substance can be collected in a state that can be directly applied to cell manipulation such as gene transfer, and a physiologically active substance can be directly collected from a biological sample such as a cell. It is an object of the present invention to provide a physiologically active substance collection device and a physiologically active substance collection method that can be collected.

上記目的を達成するために、本発明は以下の手段を提供する。
本発明は、生理活性物質を内在する細胞を含む液体または生理活性物質を含む液体を保持する液体保持部に、前記液体に接触状態に設けられた第１電極と、前記液体保持部に対して相対的に移動可能に設けられる第２電極と、該第２電極を前記細胞または液体に接触する位置と、該液体から離間する位置との間で液体保持部に対して相対的に移動させる移動手段と、前記細胞または液体に挿入された第２電極に対して、前記第１電極の電位を相対的な基準として前記生理活性物質の電荷と逆極性にバイアスされた脈流電圧を加える電源とを備える生理活性物質採取装置を提供する。 In order to achieve the above object, the present invention provides the following means.
The present invention relates to a liquid holding unit for holding a liquid containing cells containing a physiologically active substance or a liquid containing a physiologically active substance, a first electrode provided in contact with the liquid, and the liquid holding part A movement for moving the second electrode relative to the liquid holding portion between a second electrode provided so as to be relatively movable, a position where the second electrode is in contact with the cell or the liquid, and a position where the second electrode is separated from the liquid. And a power supply for applying a pulsating voltage biased to a polarity opposite to the charge of the physiologically active substance with respect to the second electrode inserted into the cell or liquid with respect to the potential of the first electrode as a relative reference. A physiologically active substance collecting device is provided.

上記発明においては、前記生理活性物質が、ＤＮＡ、ＲＮＡまたはタンパク質であることが好ましい。
また、上記発明においては、前記生理活性物質が、負の電荷を有するＤＮＡであることとしてもよい。 In the said invention, it is preferable that the said bioactive substance is DNA, RNA, or protein.
In the above invention, the physiologically active substance may be DNA having a negative charge.

また、上記発明においては、前記電源が前記第２電極に脈流電圧を加えたままの状態で、前記移動手段が前記第２電極を液体保持部に対して相対的に移動させることとしてもよい。
また、本発明は、第１電極が接触状態に配された、生理活性物質を内在する細胞または生理活性物質を含む液体に、第２電極を挿入する工程と、該第２電極に対し、前記第１電極の電位を相対的な基準として前記生理活性物質の電荷と逆極性にバイアスした脈流電圧を加える工程と、該第２電極と前記液体とを離間する位置まで相対的に移動させる工程とを備える生理活性物質採取方法を提供する。 Further, in the above invention, the moving means may move the second electrode relative to the liquid holding portion while the power supply is still applying a pulsating voltage to the second electrode. .
The present invention also includes a step of inserting the second electrode into a cell containing a physiologically active substance or a liquid containing the physiologically active substance, the first electrode being placed in contact with the second electrode, Applying a pulsating voltage biased in the opposite polarity to the charge of the physiologically active substance using the potential of the first electrode as a relative reference, and relatively moving the second electrode and the liquid to a position away from each other And a method for collecting a physiologically active substance.

本発明によれば、遺伝子導入等の細胞操作にそのまま適用可能な状態で生理活性物質を採取でき、細胞等の生体試料からも直接的に生理活性物質を採取することができるという効果を奏する。   According to the present invention, a physiologically active substance can be collected in a state that can be directly applied to cell manipulation such as gene transfer, and a physiologically active substance can be directly collected from a biological sample such as a cell.

以下、本発明の第１の実施形態に係る生理活性物質採取装置１および生理活性物質採取方法について、図１〜図８を参照して説明する。
本実施形態に係る生理活性物質採取装置１は、例えば、図１に示されるように、細胞を観察するための顕微鏡装置２に備えられる。 Hereinafter, a physiologically active substance collection device 1 and a physiologically active substance collection method according to a first embodiment of the present invention will be described with reference to FIGS.
A physiologically active substance collecting apparatus 1 according to the present embodiment is provided in a microscope apparatus 2 for observing cells, for example, as shown in FIG.

顕微鏡装置２は、図１に示されるように、遺伝子等の生理活性物質を分散させた培養液内に細胞を収容した容器３を載置するステージ４と、該ステージ４上の細胞を照明する照明装置５と、細胞において反射あるいは透過した光、あるいは細胞から発生した蛍光を観察する観察装置６と、ステージ４を駆動制御するステージコントローラ７と、照明装置５および観察装置６を制御する顕微鏡コントローラ８と、観察装置６により得られた画像を処理する画像処理装置９と、該画像処理装置９により処理された画像を表示する表示装置１０とを備えている。   As shown in FIG. 1, the microscope apparatus 2 illuminates a stage 4 on which a container 3 containing cells is placed in a culture solution in which a physiologically active substance such as a gene is dispersed, and cells on the stage 4 Illumination device 5, observation device 6 for observing light reflected or transmitted in cells, or fluorescence generated from cells, stage controller 7 for driving and controlling stage 4, and microscope controller for controlling illumination device 5 and observation device 6 8, an image processing device 9 that processes an image obtained by the observation device 6, and a display device 10 that displays an image processed by the image processing device 9.

前記照明装置５には、細胞に対して、観察装置６とは反対側から照明光を照射する透過照明光源５Ａおよび、該透過照明光源５Ａから発せられた照明光を細胞に集光するコンデンサレンズ５Ｃと、細胞に対して観察装置６と同一方向から照明光を照射する落射照明光源５Ｂとが備えられている。
また、観察装置６には、図示しない対物レンズを含む観察光学系と、該観察光学系を介した細胞からの光を撮像して画像を取得するＣＣＤ素子６Ａと、細胞からの光を直接観察する接眼レンズ６Ｂとが備えられている。 The illumination device 5 includes a transmission illumination light source 5A that irradiates the cells with illumination light from the side opposite to the observation device 6, and a condenser lens that collects the illumination light emitted from the transmission illumination light source 5A on the cells. 5C and an epi-illumination light source 5B that irradiates the cells with illumination light from the same direction as the observation device 6 are provided.
The observation device 6 includes an observation optical system including an objective lens (not shown), a CCD element 6A that captures an image of light from the cell via the observation optical system, and directly observes the light from the cell. Eyepiece 6B.

本実施形態に係る生理活性物質採取装置１は、図２に示されるように、ステージ４上の容器３近傍に配置されるＤＮＡ溶液保持部１１に設けられた第１電極１２と、該第１電極１２の上方に対向して設けられた第２電極１３と、該第２電極１３を駆動する駆動ユニット（移動手段）１４と、第２電極１３に脈流電圧を加える電気信号発生回路（電源）１５とを備えている。   As shown in FIG. 2, the physiologically active substance collection device 1 according to the present embodiment includes a first electrode 12 provided in a DNA solution holding unit 11 disposed in the vicinity of a container 3 on a stage 4, and the first electrode 12. A second electrode 13 provided facing the upper side of the electrode 12, a drive unit (moving means) 14 for driving the second electrode 13, and an electric signal generating circuit (power supply for applying a pulsating voltage to the second electrode 13 15).

ＤＮＡ溶液保持部１１は、図２に示されるように、プレート１１Ａと、該プレート１１Ａ表面に敷設されたＰｔ膜からなる第１電極１２と、ＤＮＡ溶液Ｌの保持される保持領域以外の領域のＰｔ膜を被覆するように設けられた撥水膜１１Ｂとを備えている。これにより、ＤＮＡ溶液保持部１１にＤＮＡ溶液Ｌを載せると、撥水膜１１Ｂが設けられていない保持領域に露出する第１電極１２に接触状態にＤＮＡ溶液Ｌが保持されるようになっている。   As shown in FIG. 2, the DNA solution holding unit 11 includes a plate 11A, a first electrode 12 made of a Pt film laid on the surface of the plate 11A, and regions other than the holding region where the DNA solution L is held. And a water repellent film 11B provided so as to cover the Pt film. Thus, when the DNA solution L is placed on the DNA solution holding unit 11, the DNA solution L is held in contact with the first electrode 12 exposed in the holding region where the water repellent film 11B is not provided. .

前記第２電極１３は、例えば、シリコン基板１６の表面に導電膜１３Ａを製膜することにより構成されている。第２電極１３は、カンチレバー状に形成されたシリコン基板１６の自由端に突出状態に設けられ、ＤＮＡ溶液Ｌ内に進入可能な針部１７を備えている。該針部１７にも導電膜１３Ａが設けられている。   The second electrode 13 is configured, for example, by forming a conductive film 13A on the surface of the silicon substrate 16. The second electrode 13 is provided in a protruding state at the free end of the silicon substrate 16 formed in a cantilever shape, and includes a needle portion 17 that can enter the DNA solution L. The needle portion 17 is also provided with a conductive film 13A.

前記駆動ユニット１４は、Ｘ，Ｙ，Ｚ方向に針部１７を移動させる、例えば、３軸の直線移動機構を備えている。駆動ユニット１４には、該駆動ユニット１４を制御する駆動制御装置１８が接続されている。駆動制御装置１８は、駆動ユニット１４を制御する駆動ユニット制御回路１８Ａおよび前記電気信号発生回路１５を備えている。
電気信号発生回路１５は、前記第１電極１２と第２電極１３との間に、図３に示されるような正のバイアスを有する高周波電圧を加えるようになっている。 The drive unit 14 includes, for example, a triaxial linear movement mechanism that moves the needle portion 17 in the X, Y, and Z directions. A drive control device 18 that controls the drive unit 14 is connected to the drive unit 14. The drive control device 18 includes a drive unit control circuit 18 </ b> A that controls the drive unit 14 and the electric signal generation circuit 15.
The electric signal generation circuit 15 applies a high-frequency voltage having a positive bias as shown in FIG. 3 between the first electrode 12 and the second electrode 13.

このように構成された本実施形態に係る生理活性物質採取装置１を用いた生理活性物質採取方法について以下に説明する。
本実施形態に係る生理活性物質採取装置１を用いてＤＮＡ溶液Ｌ内のＤＮＡを採取するには、ＤＮＡ溶液保持部１１の保持領域にＤＮＡ溶液Ｌを保持して、ステージ４近傍に配置し、駆動ユニット制御回路１８Ａの作動により駆動ユニット１４を作動させて第２電極１３を移動させる。そして、第２電極１３先端の針部１７がＤＮＡ溶液Ｌ内に挿入された状態で駆動ユニット１４を停止し、駆動制御装置１８に設けられた電気信号発生回路１８Ｂを作動させる。 A physiologically active substance collection method using the physiologically active substance collection device 1 according to this embodiment configured as described above will be described below.
In order to collect DNA in the DNA solution L using the physiologically active substance collecting apparatus 1 according to the present embodiment, the DNA solution L is held in the holding region of the DNA solution holding unit 11 and arranged in the vicinity of the stage 4. The drive unit 14 is operated by the operation of the drive unit control circuit 18A to move the second electrode 13. Then, the drive unit 14 is stopped in a state where the needle portion 17 at the tip of the second electrode 13 is inserted into the DNA solution L, and the electric signal generation circuit 18B provided in the drive control device 18 is operated.

電気信号発生回路１８Ｂの作動により、第１電極１２と第２電極１３との間に正のバイアスの高周波電圧が加えられる。ＤＮＡ溶液Ｌ内のＤＮＡは、負の電荷を有しているので、これにより、ＤＮＡはＤＮＡ溶液Ｌ内に挿入されている第２電極１３の針部１７のみに集められる。   By the operation of the electric signal generation circuit 18B, a positive bias high frequency voltage is applied between the first electrode 12 and the second electrode 13. Since the DNA in the DNA solution L has a negative charge, the DNA is thereby collected only in the needle portion 17 of the second electrode 13 inserted in the DNA solution L.

ＤＮＡ溶液Ｌ内に２つの電極からなるＤＮＡピンセットを刺し入れて、両電極間にＤＮＡを回収する従来の方法と比較すると、ＤＮＡを一方の第２電極１３のみに回収して、その後の操作を容易にすることができる。すなわち、第２電極１３を構成する針部１７の先端に付着した状態にＤＮＡを回収できる。したがって、ステージ４上に載置して顕微鏡観察下に配した細胞に、ＤＮＡが保持された針部１７をそのまま刺し入れて、核内に導入することができる。   Compared with the conventional method in which DNA tweezers composed of two electrodes are inserted into the DNA solution L and DNA is recovered between both electrodes, the DNA is recovered only on one of the second electrodes 13, and the subsequent operation is performed. Can be easily. That is, the DNA can be recovered in a state of being attached to the tip of the needle part 17 constituting the second electrode 13. Therefore, the needle part 17 holding the DNA can be inserted into a cell placed on the stage 4 and observed under a microscope, and introduced into the nucleus.

また、本実施形態に係る生理活性物質採取装置１によれば、第１電極１２と第２電極１３との間に加える電圧を高周波電圧とすることにより、電流の実効値を抑制し、第２電極１３の表面における反応や、ＤＮＡ溶液Ｌ中の対流を防止して、効果的に第２電極１３の表面にＤＮＡを回収することができる。   In addition, according to the physiologically active substance collecting device 1 according to the present embodiment, the effective value of the current is suppressed by setting the voltage applied between the first electrode 12 and the second electrode 13 as a high-frequency voltage. Reaction on the surface of the electrode 13 and convection in the DNA solution L can be prevented, and DNA can be effectively recovered on the surface of the second electrode 13.

ここで、図４に、本実施形態に係る生理活性物質採取装置１を用いて、ＤＮＡ溶液Ｌ中のＤＮＡを第２電極１３に回収した実施例を示す。予め蛍光標識したＤＮＡを分散させたＤＮＡ溶液Ｌ内に第２電極１３を差し込んで高周波電圧（１ＭＨｚ、２．５Ｖ、バイアス２．５Ｖ）を加え、蛍光顕微鏡により観察した結果、図４に示されるように、第２電極１３を差し込んだ右端部分の輝度が大きくなり、ＤＮＡが回収されていることがわかる。   Here, FIG. 4 shows an example in which the DNA in the DNA solution L is collected on the second electrode 13 by using the physiologically active substance collecting apparatus 1 according to the present embodiment. The second electrode 13 is inserted into the DNA solution L in which the fluorescently labeled DNA is dispersed in advance, a high-frequency voltage (1 MHz, 2.5 V, bias 2.5 V) is applied, and the result of observation with a fluorescence microscope is shown in FIG. Thus, it can be seen that the luminance at the right end portion into which the second electrode 13 is inserted is increased, and DNA is recovered.

また、このようにして電圧を加えた第２電極１３をＤＮＡ溶液Ｌから引き上げた後に、第２電極１３を蛍光観察した結果の輝度値は、図５に示されるように、電圧をかけずにＤＮＡ溶液Ｌに刺し入れて引き上げた第２電極１３と比較して２倍以上の値を示していることがわかる。
そして、このようにしてＤＮＡを付着させた第２電極１３を、そのまま、図６に矢印で示す細胞Ａに刺し入れて遺伝子導入を行い、２４時間培養した結果、図７に位相差観察像により示されるように細胞***した細胞Ａのうち、遺伝子導入を行った細胞Ａの遺伝子が発現したことが、図８に蛍光観察像により示されるように確認された。 Further, after the second electrode 13 to which voltage is applied in this way is pulled up from the DNA solution L, the luminance value as a result of fluorescence observation of the second electrode 13 is as shown in FIG. It can be seen that the value is twice or more that of the second electrode 13 inserted into the DNA solution L and pulled up.
Then, the second electrode 13 to which the DNA was attached in this way was inserted into the cell A indicated by the arrow in FIG. 6 as it was, and the gene was introduced. After culturing for 24 hours, the phase difference observation image is shown in FIG. As shown in FIG. 8, it was confirmed that the gene of the cell A into which the gene had been introduced was expressed among the cells A that had undergone cell division as shown by the fluorescence observation image in FIG.

なお、本実施形態に係る生理活性物質採取装置１においては、ＤＮＡを分散させたＤＮＡ溶液Ｌ内に第２電極１３を挿入してＤＮＡを回収する例を説明したが、これに代えて、第２電極１３を細胞Ａ内に挿入して細胞Ａ内の生理活性物質を採取することとしてもよい。   In the physiologically active substance collecting apparatus 1 according to the present embodiment, the example in which the second electrode 13 is inserted into the DNA solution L in which the DNA is dispersed and the DNA is collected has been described. The bioactive substance in the cell A may be collected by inserting the two electrodes 13 into the cell A.

この場合、顕微鏡観察下において細胞Ａの位置を確認しながら第２電極１３を挿入する必要があるため、プレート１１Ａとしてガラス基板、第１電極１２として透明電極（ＩＴＯ電極）を用いた細胞保持部１１上において細胞Ａを培養しておくことが好ましい。そして、上記と同様にして、細胞Ａ内に第２電極１３を挿入して、正のバイアスをかけた高周波電圧を加えることにより、細胞Ａ内の生理活性物質を効率的に回収することができる。
また、透明電極に代えて、第１電極１２として不透明な電極を採用し、照明光が通過する光路を避けて配置することとしてもよい。 In this case, since it is necessary to insert the second electrode 13 while confirming the position of the cell A under the microscope observation, the cell holding unit using a glass substrate as the plate 11A and a transparent electrode (ITO electrode) as the first electrode 12 11, cell A is preferably cultured in advance. In the same manner as described above, the physiologically active substance in the cell A can be efficiently recovered by inserting the second electrode 13 into the cell A and applying a positive biased high-frequency voltage. .
Further, an opaque electrode may be adopted as the first electrode 12 instead of the transparent electrode, and the first electrode 12 may be disposed avoiding the optical path through which the illumination light passes.

本発明の第１の実施形態に係る生理活性物質採取装置を示す全体構成図である。1 is an overall configuration diagram showing a physiologically active substance collection device according to a first embodiment of the present invention. 図１の生理活性物質採取装置の電極近傍の構造を示す拡大図である。It is an enlarged view which shows the structure of the electrode vicinity of the physiologically active substance collection apparatus of FIG. 図１の生理活性物質採取装置の電極に加える電圧波形例を示す図である。It is a figure which shows the voltage waveform example applied to the electrode of the physiologically active substance collection apparatus of FIG. 図１の生理活性物質採取装置の電極の周囲にＤＮＡが集まっている様子を示す顕微鏡写真である。It is a microscope picture which shows a mode that DNA has gathered around the electrode of the physiologically active substance collection apparatus of FIG. 図４の電極に高周波電圧をかけた場合とかけない場合とのＤＮＡの回収量を比較して示すグラフである。5 is a graph showing a comparison of the amount of DNA recovered when a high frequency voltage is applied to the electrode of FIG. 4 and when it is not applied. 図４のように電極に回収されたＤＮＡを導入した細胞を示す顕微鏡写真である。It is a microscope picture which shows the cell which introduce | transduced DNA collect | recovered by the electrode like FIG. 図６の細胞を２４時間培養した結果の位相差観察像を示す顕微鏡写真である。It is a microscope picture which shows the phase-contrast observation image as a result of culturing the cell of FIG. 6 for 24 hours. 図６の細胞を２４時間培養した結果の蛍光観察像を示す顕微鏡写真である。It is a microscope picture which shows the fluorescence observation image as a result of culturing the cell of FIG. 6 for 24 hours.

符号の説明Explanation of symbols

Ａ 細胞
Ｌ ＤＮＡ溶液（液体）
１ 生理活性物質採取装置
１１ ＤＮＡ溶液保持部（液体保持部）
１２ 第１電極
１３ 第２電極
１４ 駆動ユニット（移動手段）
１５ 電気信号発生回路（電源） A cell L DNA solution (liquid)
1 Bioactive substance collection device 11 DNA solution holding part (liquid holding part)
12 1st electrode 13 2nd electrode 14 Drive unit (moving means)
15 Electric signal generation circuit (power supply)

Claims (5)

生理活性物質を内在する細胞を含む液体または生理活性物質を含む液体を保持する液体保持部に、前記液体に接触状態に設けられた第１電極と、
前記液体保持部に対して相対的に移動可能に設けられる第２電極と、
該第２電極を前記細胞または液体に接触する位置と、該液体から離間する位置との間で液体保持部に対して相対的に移動させる移動手段と、
前記細胞または液体に挿入された第２電極に対して、前記第１電極の電位を相対的な基準として前記生理活性物質の電荷と逆極性にバイアスされた脈流電圧を加える電源とを備える生理活性物質採取装置。 A first electrode provided in contact with the liquid in a liquid holding unit that holds a liquid containing cells containing a physiologically active substance or a liquid containing a physiologically active substance;
A second electrode provided to be movable relative to the liquid holding unit;
Moving means for moving the second electrode relative to the liquid holding portion between a position in contact with the cell or liquid and a position away from the liquid;
A physiology provided with a power source for applying a pulsating voltage biased to a polarity opposite to the charge of the physiologically active substance with respect to the second electrode inserted into the cell or the liquid, using the potential of the first electrode as a relative reference. Active substance collection device. 前記生理活性物質が、ＤＮＡ、ＲＮＡまたはタンパク質である請求項１に記載の生理活性物質採取装置。   The physiologically active substance collecting apparatus according to claim 1, wherein the physiologically active substance is DNA, RNA, or protein. 前記生理活性物質が、負の電荷を有するＤＮＡである請求項２に記載の生理活性物質採取装置。   The physiologically active substance collecting device according to claim 2, wherein the physiologically active substance is DNA having a negative charge. 前記電源が前記第２電極に脈流電圧を加えたままの状態で、前記移動手段が前記第２電極を液体保持部に対して相対的に移動させる請求項１に記載の生理活性物質採取装置。   2. The physiologically active substance collection device according to claim 1, wherein the moving means moves the second electrode relative to the liquid holding portion while the power supply is still applying a pulsating voltage to the second electrode. . 第１電極が接触状態に配された、生理活性物質を内在する細胞または生理活性物質を含む液体に、第２電極を挿入する工程と、
該第２電極に対し、前記第１電極の電位を相対的な基準として前記生理活性物質の電荷と逆極性にバイアスした脈流電圧を加える工程と、
該第２電極と前記液体とを離間する位置まで相対的に移動させる工程とを備える生理活性物質採取方法。 Inserting the second electrode into a cell containing the physiologically active substance or a liquid containing the physiologically active substance, the first electrode being placed in contact;
Applying, to the second electrode, a pulsating voltage biased to a polarity opposite to the charge of the physiologically active substance with respect to the potential of the first electrode as a relative reference;
And a step of relatively moving the second electrode and the liquid to a position where they are separated from each other.
JP2006150352A 2006-05-30 2006-05-30 Device for collecting physiologically active substance and method for the same Withdrawn JP2007319035A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011523064A (en) * 2008-06-06 2011-08-04 ユニバーシティー・オブ・ワシントン Method and system for concentrating particles from solution
US8940092B1 (en) 2008-10-27 2015-01-27 University Of Washington Through Its Center For Commercialization Hybrid fibers, devices using hybrid fibers, and methods for making hybrid fibers

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2011523064A (en) * 2008-06-06 2011-08-04 ユニバーシティー・オブ・ワシントン Method and system for concentrating particles from solution
US8632669B2 (en) 2008-06-06 2014-01-21 University Of Washington Method and system for concentrating particles from a solution
US9097664B2 (en) 2008-06-06 2015-08-04 University Of Washington Method and system for concentrating particles from a solution
US9518956B2 (en) 2008-06-06 2016-12-13 University Of Washington Particle concentration system
US8940092B1 (en) 2008-10-27 2015-01-27 University Of Washington Through Its Center For Commercialization Hybrid fibers, devices using hybrid fibers, and methods for making hybrid fibers

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