JPH04250332A - Trapping device for organism-related material and observing method for organism-related material using the same - Google Patents
Trapping device for organism-related material and observing method for organism-related material using the sameInfo
- Publication number
- JPH04250332A JPH04250332A JP859891A JP859891A JPH04250332A JP H04250332 A JPH04250332 A JP H04250332A JP 859891 A JP859891 A JP 859891A JP 859891 A JP859891 A JP 859891A JP H04250332 A JPH04250332 A JP H04250332A
- Authority
- JP
- Japan
- Prior art keywords
- related substance
- comb
- pair
- biologically
- electric field
- Prior art date
- Legal status (The legal status 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 status listed.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims description 25
- 239000000463 material Substances 0.000 title abstract 7
- 239000000523 sample Substances 0.000 claims abstract description 79
- 230000005684 electric field Effects 0.000 claims abstract description 25
- 239000000126 substance Substances 0.000 claims description 83
- 239000000725 suspension Substances 0.000 claims description 6
- 210000000056 organ Anatomy 0.000 abstract 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 11
- 238000010586 diagram Methods 0.000 description 11
- 229910002804 graphite Inorganic materials 0.000 description 11
- 239000010439 graphite Substances 0.000 description 11
- 239000012620 biological material Substances 0.000 description 4
- 230000005686 electrostatic field Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 102000029749 Microtubule Human genes 0.000 description 3
- 108091022875 Microtubule Proteins 0.000 description 3
- FCZCIXQGZOUIDN-UHFFFAOYSA-N ethyl 2-diethoxyphosphinothioyloxyacetate Chemical compound CCOC(=O)COP(=S)(OCC)OCC FCZCIXQGZOUIDN-UHFFFAOYSA-N 0.000 description 3
- 210000004688 microtubule Anatomy 0.000 description 3
- 230000005641 tunneling Effects 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 102000008946 Fibrinogen Human genes 0.000 description 2
- 108010049003 Fibrinogen Proteins 0.000 description 2
- 238000002441 X-ray diffraction Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229940012952 fibrinogen Drugs 0.000 description 2
- 230000003436 cytoskeletal effect Effects 0.000 description 1
- 210000003963 intermediate filament Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
Description
【０００１】0001
【産業上の利用分野】本発明は、ＤＮＡ、バクテリア、
フィブリノーゲンなどの生体関連物質の捕捉装置とそれ
を用いた観察方法に関するものである。[Industrial Application Field] The present invention is applicable to DNA, bacteria,
This invention relates to a device for capturing bio-related substances such as fibrinogen and an observation method using the same.
【０００２】0002
【従来の技術】近年、走査形トンネル顕微鏡（Ｓｃａｎ
ｎｉｎｇ Ｔｕｎｎｅｌｉｎｇ Ｍｉｃｒｏｓｃｏ
ｐｅ、以下ＳＴＭという）や原子間顕微鏡（Ａｔｏｍｉ
ｃ Ｆｏｒｃｅ Ｍｉｃｒｏｓｃｏｐｅ、以下ＡＦ
Ｍという）を用いて、サブナノメータからμｍのオーダ
で様々な物体の観察が行われている。表面界面物性等の
研究では、試料として準備された固体表面が直接観察さ
れる。
それに対し、生体関連物質などの微小な試料を観察する
場合は、試料をまずプレパラートとして準備したグラフ
ァイトの上に固定し、次に観察が行われる。[Prior Art] In recent years, scanning tunneling microscopes (Scan
ning Tunneling Microsco
pe (hereinafter referred to as STM) and atomic microscope (Atomi
c Force Microscope, hereinafter AF
A variety of objects have been observed on the order of subnanometers to micrometers by using the micrometer (referred to as M). In research on surface and interfacial properties, solid surfaces prepared as samples are directly observed. On the other hand, when observing a minute sample such as a biological substance, the sample is first fixed on graphite prepared as a slide and then observed.
【０００３】図６はかかる従来の試料をグラファイト上
に固定して観察する状態を示す斜視図である。この図に
示すように、プレパラートとしてのグラファイト１上に
試料２を載置して、探針３をグラファイト１上に移動さ
せて観察を行うようにしていた。このように、従来の方
法ではプレパラートとして準備したグラファイト１の上
に試料を固定して観察を行っている。その場合、試料２
が固定される時の試料の姿勢や方向は制御されていない
ため、図６に示すように、試料２はグラファイト１上に
雑然と捕捉される。そして、生体物質の観察を行うため
のＳＴＭ、ＡＦＭは広い視野が必要とされている（例え
ば、「Ａ ｓｍａｌｌ ｓｃａｎｎｉｎｇｔｕｎｎ
ｅｌｌｎｇ ｍｉｃｒｏｓｃｏｐｅ ｗｉｔｈ
ｌａｒｇｅ ｓｃａｎ ｒａｎｇｅ ｆｏｒ
ｂｉｏｌｏｇｉｃａｌ ｓｔｕｄｉｅｓ」 ｂｙ
Ｒ．Ｅｍｃｈ，Ｐ．Ｄｅｓｃｏｕｔｓ ｅｔ ａ
ｌ，Ｊｏｕｒｎａｌ ｏｆ Ｍｉｃｒｏｓｃｏｐｙ
，Ｖｏｌ．１５２，Ｐｔ１，Ｏｃｔｏｂｅｒ １９８
８，ｐｐ．８５−９２ 参照）。FIG. 6 is a perspective view showing a state in which such a conventional sample is fixed on graphite and observed. As shown in this figure, a sample 2 was placed on graphite 1 as a preparation, and a probe 3 was moved onto the graphite 1 for observation. As described above, in the conventional method, a sample is fixed on graphite 1 prepared as a slide and observed. In that case, sample 2
Since the posture and direction of the sample when it is fixed are not controlled, the sample 2 is roughly captured on the graphite 1, as shown in FIG. STM and AFM for observing biological materials require a wide field of view (for example, "A small scanning tunnel").
ellng microscope with
large scan range for
biological studies” by
R. Emch, P. Descouts et a
l, Journal of Microscopy
, Vol. 152, Pt1, October 198
8, pp. 85-92).
【０００４】0004
【発明が解決しようとする課題】しかしながら、上記し
た従来の観察方法では、（１）試料２がプレパラートと
してのグラファイト１上に雑然と捕捉されるため、有効
な試料を広い観察領域の中から探さなければならない。
（２）また、試料２がプレパラートとしてのグラファイ
ト１上に雑然と捕捉されるため、試料２の交差などが生
じ易い。（３）その時々により捕捉状態が変わり、試料
２の同定の確度（再現性）が低い。また、更に、（４）
プレパラートとしてのグラファイト１上に載置されるた
め、試料２がそのグラファイト１上で潰れてしまう。
（５）試料を一方向からのみ観察を行うため、探針の有
効先端半径によって像が丸められる（例えば、「Ｓｃａ
ｎｎｉｎｇ ｔｕｎｎｅｌｉｎｇ ｍｉｃｒｏｓｃ
ｏｐｅ ｏｆ ｃｙｔｏｓｋｅｌｅｔａｌ ｐｒ
ｏｔｅｉｎｓ：Ｍｉｃｒｏｔｕｂｕｌｅｓ ａｎｄ
ｉｎｔｅｒｍｅｄｉａｔｅ ｆｉｌａｍｅｎｔｓ」
ｂｙ Ｓｔｕａｒｔ Ｈａｍｅｒｏｆｆｅｔ ａ
ｌ，Ｊ．Ｖａｃ．Ｓｃｉ．Ｔｅｃｈｎｏｌ．Ａ８（１）
，Ｊａｎ／Ｆｅｂ，１９９０，ｐｐ６８７−６９１参照
）といった問題があった。例えば、図７に示すような、
Ｘ線回析によった生体物質としての微小管（ｍｉｃｒｏ
ｔｕｂｕｌｅ）４のＳＴＭイメージは図８に示すように
なる。[Problems to be Solved by the Invention] However, in the conventional observation method described above, (1) the sample 2 is roughly captured on the graphite 1 as a preparation, so it is difficult to search for an effective sample from a wide observation area; There must be. (2) Furthermore, since the sample 2 is roughly captured on the graphite 1 as a preparation, the samples 2 are likely to cross each other. (3) The capture state changes from time to time, and the accuracy (reproducibility) of identifying sample 2 is low. Furthermore, (4)
Since the sample 2 is placed on the graphite 1 as a preparation, the sample 2 is crushed on the graphite 1. (5) Since the sample is observed from only one direction, the image is rounded by the effective tip radius of the probe (for example, “Sca
nnning tunneling microsc
open of cytoskeletal pr
oteins: Microtubules and
"intermediate filaments"
by Stuart Hameroffet a
l, J. Vac. Sci. Technol. A8 (1)
, Jan/Feb, 1990, pp. 687-691). For example, as shown in FIG.
Microtubules as a biological material by X-ray diffraction
The STM image of tube) 4 is shown in FIG.
【０００５】本発明は、上記問題点を除去するために、
電界をかけることにより、生体関連物質としての試料の
マニピュレーションと捕捉を行い、迅速、かつ的確に観
察することができ、しかも試料をより本来の姿に近い形
で観察可能な生体関連物質の捕捉装置とそれを用いた観
察方法を提供することを目的とする。[0005] In order to eliminate the above problems, the present invention has the following features:
A capture device for bio-related substances that can manipulate and capture samples as bio-related substances by applying an electric field, and observe them quickly and accurately, as well as allowing the specimen to be observed in a form closer to its original form. The purpose is to provide an observation method using it.
【０００６】[0006]
【課題を解決するための手段】本発明は、上記目的を達
成するために、（Ａ）生体関連物質の捕捉装置において
、プレパラートと、該プレパラートに形成される対向し
た一対の櫛形電極と、該一対の櫛形電極間に電界をかけ
、該櫛形電極の歯の間に生体関連物質を捕捉する手段と
を設けるようにしたものである。[Means for Solving the Problems] In order to achieve the above object, the present invention provides (A) a biologically related substance trapping device that includes a preparation, a pair of opposed comb-shaped electrodes formed on the preparation; A means is provided for applying an electric field between a pair of comb-shaped electrodes and trapping biological substances between the teeth of the comb-shaped electrodes.
【０００７】また、前記プレパラートの生体関連物質の
捕捉部に凹溝を形成し、前記生体関連物質を吊り橋状に
捕捉するようにしたものである。更に、前記凹溝の底部
に複数の電極を配置し、該電極の電位を制御することに
より前記生体関連物質を変位させるようにしたものであ
る。
（Ｂ）生体関連物質の観察方法において、プレパラート
に形成される対向した一対の櫛形電極間に電界をかけ、
該櫛形電極の歯の間に生体関連物質を捕捉し、該捕捉さ
れた生体関連物質を探針を走査して観察するようにした
ものである。
（Ｃ）生体関連物質の観察方法において、プレパラート
の捕捉部に凹溝を形成し、該凹溝の両側に対向した一対
の櫛形電極間に電界をかけ、該櫛形電極の歯の間に生体
関連物質を吊り橋状に捕捉し、該捕捉された生体関連物
質を探針を走査して観察するようにしたものである。
（Ｄ）生体関連物質の観察方法において、プレパラート
の捕捉部に凹溝を形成し、該凹溝の両側に対向した一対
の櫛形電極間に電界をかけ、該櫛形電極の歯の間に生体
関連物質を吊り橋状に捕捉し、前記凹溝の底部に電極を
設け、該電極の電位を制御し、該捕捉された生体関連物
質を回転させながら探針を走査して観察するようにした
ものである。
（Ｅ）生体関連物質の捕捉装置において、三次元空間に
対向した一対の捕捉用針と、該一対の捕捉用針間に電界
をかけ、生体関連物質を捕捉する手段とを設け、該生体
関連物質を三次元的に観察するようにしたものである。[0007]Furthermore, a concave groove is formed in the bio-related substance trapping portion of the preparation, so that the bio-related substance is captured in the form of a suspension bridge. Furthermore, a plurality of electrodes are arranged at the bottom of the groove, and the biologically related substance is displaced by controlling the potential of the electrodes. (B) In a method for observing biological substances, applying an electric field between a pair of opposing comb-shaped electrodes formed on a preparation,
A biological substance is captured between the teeth of the comb-shaped electrode, and the captured biological substance is observed by scanning the probe. (C) In a method for observing biologically related substances, a groove is formed in the capture portion of the preparation, an electric field is applied between a pair of comb-shaped electrodes facing each other on both sides of the groove, and biologically related substances are generated between the teeth of the comb-shaped electrodes. A substance is captured in the shape of a suspension bridge, and the captured biologically related substance is observed by scanning with a probe. (D) In the method for observing biologically related substances, a groove is formed in the trapping part of the preparation, an electric field is applied between a pair of comb-shaped electrodes facing each other on both sides of the groove, and biologically related substances are applied between the teeth of the comb-shaped electrodes. The substance is captured in the form of a suspension bridge, an electrode is provided at the bottom of the groove, the potential of the electrode is controlled, and the probe is scanned and observed while rotating the captured biological substance. be. (E) A capture device for biologically related substances, which includes a pair of capturing needles facing each other in three-dimensional space and means for applying an electric field between the pair of capturing needles to capture the biologically related substances. This allows for three-dimensional observation of matter.
【０００８】また、前記生体関連物質を中心としてラジ
アル電極を形成するようにしたものである。
（Ｆ）生体関連物質の観察方法において、空間に対向し
た一対の捕捉用針間に電界をかけ、生体関連物質を捕捉
し、該生体関連物質を探針により三次元的に観察するよ
うにしたものである。
（Ｇ）生体関連物質の観察方法において、空間に対向し
た一対の捕捉用針間に電界をかけ、生体関連物質を捕捉
し、前記生体関連物質を中心としたラジアル電極を設け
、該ラジアル電極への電位の印加により、生体関連物質
を回転させながら探針により三次元的に観察するように
したものである。[0008] Furthermore, the radial electrode is formed mainly of the biologically related substance. (F) In a method for observing biological substances, an electric field is applied between a pair of trapping needles facing each other in space to capture biological substances, and the biological substances are observed three-dimensionally using a probe. It is something. (G) In a method for observing biological substances, an electric field is applied between a pair of trapping needles facing each other in space to capture the biological substance, a radial electrode is provided around the biological substance, and the radial electrode is connected to the biological substance. By applying an electric potential, biological substances are rotated and observed three-dimensionally with a probe.
【０００９】[0009]
【作用】本発明によれば、（１）試料捕捉面（プレパラ
ート面）に沿って電界をかけ、生体関連物質である試料
を列状に捕捉する。従って、同一試料が整然と列状に捕
捉されることにより、試料の観察結果の再現性が検証さ
れ、試料の同定の確度の向上を図ることができる。
（２）生体関連物質である試料の捕捉のための針を一対
用意し、その間に静電界をかけることによって線状の試
料を一対の針の間に捕捉する。それにより、プレパラー
トを用いずに試料の観察を行うことが可能になり、試料
をより本来の姿に近い形で観察することができる。
（３）捕捉用針の間に捕捉された試料を回転する静電界
をかけて回転させる。それにより、観察用探針の先端有
効半径による像の丸まりの影響を低減することができる
。[Function] According to the present invention, (1) an electric field is applied along the sample capture surface (preparation surface) to capture samples, which are biological substances, in a row. Therefore, by capturing the same sample in an orderly row, the reproducibility of the sample observation results can be verified, and the accuracy of sample identification can be improved. (2) A pair of needles for capturing a sample, which is a biological substance, is prepared, and a linear sample is captured between the pair of needles by applying an electrostatic field between them. Thereby, it becomes possible to observe the sample without using a preparation, and the sample can be observed in a form closer to its original appearance. (3) A rotating electrostatic field is applied to rotate the sample captured between the capturing needles. Thereby, the influence of image rounding due to the effective radius of the tip of the observation probe can be reduced.
【００１０】従って、生体関連物質の捕捉を迅速、かつ
的確に行うことができ、益々増大する生体関連物質の観
察の信頼性を向上させると共に、その自動化を図ること
ができる。[0010] Therefore, biological substances can be captured quickly and accurately, and the reliability of the observation of biological substances, which is increasingly increasing, can be improved and the observation can be automated.
【００１１】[0011]
【実施例】以下、本発明の実施例について図面を参照し
ながら詳細に説明する。図１は本発明の第１の実施例を
示す生体関連物質の観察方法を示す図であり、図１（ａ
）はそれに用いる捕捉用プレパラートの斜視図、図１（
ｂ）はその捕捉用プレパラート上に試料を捕捉し、観察
を行う状態を示す斜視図である。Embodiments Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings. FIG. 1 is a diagram illustrating a method for observing biological substances according to a first embodiment of the present invention, and FIG.
) is a perspective view of the capture preparation used for it, and Figure 1 (
b) is a perspective view showing a state in which a sample is captured on the capture preparation and observed.
【００１２】ここで、捕捉用プレパラートは、半導体集
積回路製造技術を応用して製造され、微細な対向した櫛
形電極が形成される。まず、生体関連物質（ＤＮＡ、バ
クテリア、フィブリノーゲン等）を観察する場合、図１
（ａ）に示すように、捕捉用プレパラート１１上には捕
捉用櫛形電極１２と捕捉用櫛形電極１３とが対向するよ
うに形成され、それらの櫛形電極１２と１３間に端子１
５と１６を介して電圧を印加し、対向する櫛形電極の歯
１２ａと１３ａ間に電界（例えば、１０６ Ｖ／ｍ）を
かける。なお、ここで、印加される電圧は、通常、交流
を用いるが、直流を用いるようにしてもよい。[0012] Here, the trapping preparation is manufactured by applying semiconductor integrated circuit manufacturing technology, and fine, opposed comb-shaped electrodes are formed. First, when observing biological substances (DNA, bacteria, fibrinogen, etc.), Figure 1
As shown in (a), a trapping comb-shaped electrode 12 and a trapping comb-shaped electrode 13 are formed on the trapping preparation 11 so as to face each other, and a terminal 1 is provided between the comb-shaped electrodes 12 and 13.
A voltage is applied through 5 and 16 to create an electric field (for example 106 V/m) between opposing comb electrode teeth 12a and 13a. Note that, although alternating current is normally used as the applied voltage here, direct current may also be used.
【００１３】次に、捕捉用櫛形電極１２と１３間に電圧
が印加された状態で、生体関連物質としての試料１４を
分散させた溶液を捕捉用プレパラート１１上に導入する
。すると、図１（ｂ）に示すように、試料１４は捕捉さ
れる際、捕捉用プレパラート１１の表面の電界の影響を
受けて配列し、櫛形電極の各歯１２ａと各歯１３ａの間
に捕捉される。その後、そのまま液中で観察を行うか、
もしくは、液を取り除き、ドライな状態でＳＴＭやＡＦ
Ｍの探針１５による観察を行う。Next, while a voltage is applied between the capture comb-shaped electrodes 12 and 13, a solution in which a sample 14 as a biologically related substance is dispersed is introduced onto the capture preparation 11. Then, as shown in FIG. 1(b), when the samples 14 are captured, they are arranged under the influence of the electric field on the surface of the capture preparation 11, and are captured between each tooth 12a and each tooth 13a of the comb-shaped electrode. be done. After that, you can observe it directly in the liquid, or
Alternatively, remove the liquid and use STM or AF in a dry state.
Observation is performed using the M probe 15.
【００１４】なお、櫛形電極１２と１３間に印加される
電圧及び周波数は、捕捉される試料１４の種類や条件に
応じて適宜設定することができる。また、櫛形電極１２
と１３の歯１２ａと１３ａの先端は尖らせて、試料１４
を捕捉し易いようにすることが望ましい。図２は本発明
の第２の実施例を示す生体関連物質の捕捉装置とそれを
用いた観察方法を示す図である。Note that the voltage and frequency applied between the comb-shaped electrodes 12 and 13 can be appropriately set depending on the type and conditions of the sample 14 to be captured. In addition, the comb-shaped electrode 12
The tips of teeth 12a and 13a of sample 14 and 13 are made sharp.
It is desirable to make it easy to capture. FIG. 2 is a diagram illustrating a biological substance capture device and an observation method using the same, which is a second embodiment of the present invention.
【００１５】この図に示すように、この実施例において
は、従来の欠点である（１）試料が二次元的に潰れてし
まう。（２）一方向からのみ観察を行うため、探針の有
効先端半径によって像が丸められるなどの問題点を解決
するために、四探針型ＡＦＭを構成する。即ち、図２に
示すように、一対の捕捉用針２１と２２を対向させ、そ
れらの捕捉用針２１と２２間に端子２５と２６を介して
電圧を印加し、適当な電界をかけることによって、生体
関連物質である試料２３を捕捉する。[0015] As shown in this figure, this embodiment has the disadvantages of the conventional method (1): the sample is crushed two-dimensionally; (2) Since observation is performed only from one direction, a four-probe AFM is configured to solve the problem that the image is rounded due to the radius of the effective tip of the probe. That is, as shown in FIG. 2, a pair of capture needles 21 and 22 are placed opposite each other, and a voltage is applied between the capture needles 21 and 22 via terminals 25 and 26 to apply an appropriate electric field. , captures a sample 23 which is a biologically related substance.
【００１６】それにより、試料２３を二次元的に拘束す
ることなく捕捉することができる。そこで、探針２４を
移動させることにより、ＳＴＭ、ＡＦＭによる観察を行
うと、試料本来の形に近い形で三次元的観察を行うこと
ができる。図３は本発明の第３の実施例を示す生体関連
物質の捕捉装置とそれを用いた観察方法を示す図である
。[0016] Thereby, the sample 23 can be captured without being constrained two-dimensionally. Therefore, by moving the probe 24 and performing observation using STM or AFM, it is possible to perform three-dimensional observation in a shape close to the original shape of the sample. FIG. 3 is a diagram illustrating a biological substance capture device and an observation method using the device according to a third embodiment of the present invention.
【００１７】この図に示すように、この実施例において
は、図２の生体関連物質の捕捉装置に、更に、試料が捕
捉される方向を軸としたラジアル電極２７を配置し、該
ラジアル電極２７に試料２３を軸として回転する電圧を
印加することによって、試料２３を捕捉用針２１，２２
に捕捉された状態でその軸回りに回転させることができ
る。それにより、探針２４の先端有効半径による像の丸
まりを低減することができ、的確な三次元的観察を行う
ことができる。As shown in this figure, in this embodiment, a radial electrode 27 whose axis is in the direction in which the sample is captured is further arranged in the bio-related substance capture device of FIG. By applying a voltage that rotates around the sample 23 to
It can be rotated around its axis while being held in place. Thereby, it is possible to reduce the rounding of the image due to the effective radius of the tip of the probe 24, and it is possible to perform accurate three-dimensional observation.
【００１８】図４は本発明の第４の実施例を示す生体関
連物質の捕捉装置とそれを用いた観察方法を示す図であ
る。この図に示すように、この実施例においては、捕捉
用櫛状電極の各歯が空間にオーバーハング状態に配置さ
れている。つまり、捕捉用プレパラート３１の試料捕捉
部には凹溝３２が形成され、この凹溝３２の両側から捕
捉用櫛形電極３３の歯３３ａと捕捉用櫛形電極３４の歯
３４ａが突出し、互いに対向している。そして、これら
の捕捉用櫛形電極３３と３４間には端子３６と３７を介
して適当な電圧が印加され、捕捉用櫛形電極３３と３４
の各歯３３ａと３４ａ間に電界がかけられている。従っ
て、これらの捕捉用櫛形電極３３と３４の各歯３３ａと
３４ａ間に試料３５が二次元的に潰されることなく捕捉
される。FIG. 4 is a diagram illustrating a biological substance capture device and an observation method using the device according to a fourth embodiment of the present invention. As shown in this figure, in this embodiment, each tooth of the capture comb-shaped electrode is arranged in an overhanging state in the space. That is, a groove 32 is formed in the sample trapping part of the trapping preparation 31, and the teeth 33a of the comb-shaped trapping electrode 33 and the teeth 34a of the comb-shaped trapping electrode 34 protrude from both sides of the groove 32, and are opposed to each other. There is. An appropriate voltage is applied between these comb-shaped trapping electrodes 33 and 34 via terminals 36 and 37, and the comb-shaped trapping electrodes 33 and 34
An electric field is applied between each tooth 33a and 34a. Therefore, the sample 35 is captured between the teeth 33a and 34a of the capturing comb-shaped electrodes 33 and 34 without being crushed two-dimensionally.
【００１９】そこで、試料を分散させた溶液を凹溝３２
に導入する。すると、試料３５は捕捉される際、表面の
電界の影響を受けて配列し、櫛形電極の各歯３３ａと各
歯３４ａの間に捕捉される。その後、そのまま液中で観
察を行うか、もしくは、液を取り除き、ドライな状態で
ＳＴＭやＡＦＭの探針３８による観察を行う。図５は本
発明の第５の実施例を示す生体関連物質の捕捉装置とそ
れを用いた観察方法を示す図である。Therefore, the solution in which the sample is dispersed is poured into the groove 32.
to be introduced. Then, when the sample 35 is captured, it is arranged under the influence of the electric field on the surface and is captured between each tooth 33a and each tooth 34a of the comb-shaped electrode. Thereafter, observation is performed directly in the liquid, or the liquid is removed and observation is performed in a dry state using an STM or AFM probe 38. FIG. 5 is a diagram illustrating a biological substance capture device and an observation method using the device according to a fifth embodiment of the present invention.
【００２０】この図に示すように、この実施例において
は、図４の生体関連物質の捕捉装置に、更に、凹溝３２
の底部に複数の帯状の電極３９を形成する。そこで、こ
れらの電極３９に印加される電位を制御することによっ
て、捕捉された試料３５を一斉に回転（変位）させ、同
一の姿勢（回転角）で探針３８により、該試料３５を観
察することができる。As shown in this figure, in this embodiment, the biological substance trapping device of FIG.
A plurality of strip-shaped electrodes 39 are formed on the bottom of the electrode. Therefore, by controlling the potential applied to these electrodes 39, the captured samples 35 are rotated (displaced) all at once, and the samples 35 are observed with the probe 38 in the same posture (rotation angle). be able to.
【００２１】また、上記生体関連物質の捕捉装置により
、試料が捕捉されたことを以下のようにして確認するこ
とができる。
（１）図２及び図３に関して試料２３が捕捉用針２１，
２２間に捕捉されたことを確認する場合、捕捉用針２１
，２２にＡＦＭの探針としての力検出機能を持たせた状
態で、捕捉用針２１，２２を微小に振動させ、その振幅
が大きく変化した時を持って捕捉が行われたと見ること
ができる。
（２）捕捉用針のアライメント
図２及び図３に関して一対の捕捉用針は試料が小さいほ
ど厳密な相互のアライメントを行う必要がある。そのた
めに捕捉用針にＡＦＭ探針としての力検出機能を持たせ
る。そこで、一方の捕捉用針を用いて他方の捕捉用針を
観察することによって相互の頂点を一致させることがで
きる。これによって厳密なアライメントが可能となり、
既知の非常に微小な領域に試料を捕捉することができる
。[0021]Furthermore, it is possible to confirm that a sample has been captured by the biological substance capture device as described below. (1) Regarding FIGS. 2 and 3, the sample 23 is the capturing needle 21,
If you want to confirm that it has been captured between 22 and 22, use the capture needle 21
, 22 have a force detection function as an AFM probe, the acquisition needles 21 and 22 are slightly vibrated, and when the amplitude changes significantly, it can be determined that acquisition has occurred. . (2) Alignment of Capture Needles Regarding FIGS. 2 and 3, the smaller the sample, the more strictly the pair of capture needles must be aligned with each other. For this purpose, the capturing needle is provided with a force detection function as an AFM probe. Therefore, by observing the other capturing needle using one capturing needle, it is possible to match the mutual apexes. This allows for precise alignment,
The sample can be captured in a known, extremely small area.
【００２２】なお、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いて種々の変形が可能
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。Note that the present invention is not limited to the above-mentioned 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.
【００２３】[0023]
【発明の効果】以上、詳細に説明したように、本発明に
よれば、生体関連物質である試料の捕捉面（プレパラー
ト面）に沿って電界をかけ、線状の同一試料が列状に捕
捉されるようにしたので、同一試料が整然と列状に捕捉
されることにより、試料の観察結果の再現性が検証され
、試料の同定の確度の向上を図ることができる。Effects of the Invention As described above in detail, according to the present invention, an electric field is applied along the capture surface (preparation surface) of a sample, which is a biological substance, and the same linear sample is captured in a row. Since the same sample is captured in an orderly row, the reproducibility of the sample observation results can be verified and the accuracy of sample identification can be improved.
【００２４】また、試料の捕捉のための針を一対用意し
、その間に静電界をかけることによって線状の試料を一
対の針の間に捕捉する。それにより、プレパラートを用
いずに試料の観察を行うことが可能になり、試料をより
本来の姿に近い形で観察することができる。更に、捕捉
用針の間に捕捉された試料を回転する静電界をかけて回
転させる。それにより、観察用探針の先端有効半径によ
る像の丸まりの影響を低減することができる。Further, a pair of needles for capturing a sample is prepared, and a linear sample is captured between the pair of needles by applying an electrostatic field between them. Thereby, it becomes possible to observe the sample without using a preparation, and the sample can be observed in a form closer to its original appearance. Furthermore, a rotating electrostatic field is applied to rotate the sample captured between the capturing needles. Thereby, the influence of image rounding due to the effective radius of the tip of the observation probe can be reduced.
【００２５】従って、本発明によれば、生体関連物質の
捕捉を迅速、かつ的確に行うことができ、益々増大する
生体関連物質の観察の信頼性を向上させると共に、その
自動化を図ることができ、本発明によってもたらされる
効果は著大である。[0025] Therefore, according to the present invention, it is possible to quickly and accurately capture biological substances, improve the reliability of the observation of biological substances, which are increasingly increasing, and to automate the observation. , the effects brought about by the present invention are significant.
【図１】本発明の第１の実施例を示す生体関連物質の観
察方法を示す図である。FIG. 1 is a diagram illustrating a method for observing biological substances according to a first embodiment of the present invention.
【図２】本発明の第２の実施例を示す生体関連物質の捕
捉装置とそれを用いた観察方法を示す図である。FIG. 2 is a diagram illustrating a biologically related substance capturing device and an observation method using the same, showing a second embodiment of the present invention.
【図３】本発明の第３の実施例を示す生体関連物質の捕
捉装置とそれを用いた観察方法を示す図である。FIG. 3 is a diagram illustrating a biologically related substance capturing device and an observation method using the same according to a third embodiment of the present invention.
【図４】本発明の第４の実施例を示す生体関連物質の捕
捉装置とそれを用いた観察方法を示す図である。FIG. 4 is a diagram illustrating a bio-related substance capturing device and an observation method using the same, showing a fourth embodiment of the present invention.
【図５】本発明の第５の実施例を示す生体関連物質の捕
捉装置とそれを用いた観察方法を示す図である。FIG. 5 is a diagram illustrating a biologically related substance capturing device and an observation method using the same according to a fifth embodiment of the present invention.
【図６】従来の試料をグラファイト上に固定して観察す
る状態を示す斜視図である。FIG. 6 is a perspective view showing a state in which a conventional sample is fixed on graphite and observed.
【図７】従来のＸ線回析による生体物質としての微小管
の斜視図である。FIG. 7 is a perspective view of microtubules as a biological material obtained by conventional X-ray diffraction.
【図８】従来の生体物質のＳＴＭイメージを示す図であ
る。FIG. 8 is a diagram showing a conventional STM image of a biological material.
１１，３１ 捕捉用プレパラート１２，１３，３
３，３４ 捕捉用櫛形電極１２ａ，１３ａ，３３
ａ，３４ａ 櫛形電極の歯１４，２３，３５
試料（生体関連物質）１５，１６，２５，２６，３
６，３７ 端子１７，２４，３８ 探針
２１，２２ 捕捉用針
２７ ラジアル電極
３２ 凹溝
３９ 帯状の電極11, 31 Capture preparation 12, 13, 3
3, 34 Comb-shaped capture electrodes 12a, 13a, 33
a, 34a Comb-shaped electrode teeth 14, 23, 35
Samples (biological substances) 15, 16, 25, 26, 3
6, 37 Terminals 17, 24, 38 Probes 21, 22 Capture needle 27 Radial electrode 32 Concave groove 39 Band-shaped electrode
Claims (10)
ートに形成される対向した一対の櫛形電極と、（ｃ）該
一対の櫛形電極間に電界をかけ、該櫛形電極の歯の間に
生体関連物質を捕捉する手段とを具備する生体関連物質
の捕捉装置。Claim 1: Applying an electric field between (a) a preparation, (b) a pair of opposing comb-shaped electrodes formed on the preparation, and (c) the pair of comb-shaped electrodes, and applying an electric field between the teeth of the comb-shaped electrodes. 1. A biologically related substance capturing device, comprising: a means for capturing a biologically related substance.
捉部に凹溝を形成し、前記生体関連物質を吊り橋状に捕
捉してなる請求項１記載の生体関連物質の捕捉装置。2. The biologically related substance trapping device according to claim 1, wherein a concave groove is formed in the biologically related substance trapping portion of the preparation to trap the biologically related substance in the form of a suspension bridge.
、該電極の電位を制御することにより前記生体関連物質
を変位させることを特徴とする請求項１記載の生体関連
物質の捕捉装置。3. The biologically related substance trapping device according to claim 1, wherein a plurality of electrodes are disposed at the bottom of the groove, and the biologically related substance is displaced by controlling the potential of the electrodes. .
対の櫛形電極間に電界をかけ、該櫛形電極の歯の間に生
体関連物質を捕捉し、該捕捉された生体関連物質を探針
を走査して観察する生体関連物質の観察方法。4. Applying an electric field between a pair of opposing comb-shaped electrodes formed on a preparation, trapping a bio-related substance between the teeth of the comb-shaped electrode, and scanning the captured bio-related substance with a probe. A method for observing biologically related substances.
、該凹溝の両側に対向した一対の櫛形電極間に電界をか
け、該櫛形電極の歯の間に生体関連物質を吊り橋状に捕
捉し、該捕捉された生体関連物質を探針を走査して観察
する生体関連物質の観察方法。5. A concave groove is formed in the capture portion of the preparation, an electric field is applied between a pair of comb-shaped electrodes facing each other on both sides of the concave groove, and biologically related substances are captured between the teeth of the comb-shaped electrodes in the form of a suspension bridge. A method for observing a biologically-related substance, in which the captured biologically-related substance is observed by scanning with a probe.
、該凹溝の両側に対向した一対の櫛形電極間に電界をか
け、該櫛形電極の歯の間に生体関連物質を吊り橋状に捕
捉し、前記凹溝の底部に電極を設け、該電極の電位を制
御し、該捕捉された生体関連物質を回転させながら探針
を走査して観察する生体関連物質の観察方法。6. Forming a concave groove in the trapping portion of the preparation, applying an electric field between a pair of comb-shaped electrodes facing each other on both sides of the concave groove, and capturing biological substances between the teeth of the comb-shaped electrodes in the form of a suspension bridge. A method for observing a living body-related substance, wherein an electrode is provided at the bottom of the groove, the electric potential of the electrode is controlled, and the captured living body-related substance is scanned and observed with a probe while rotating.
針と、（ｂ）該一対の捕捉用針間に電界をかけ、生体関
連物質を捕捉する手段とを設け、（ｃ）該生体関連物質
を三次元的に観察することを特徴とする生体関連物質の
捕捉装置。7. A method comprising: (a) a pair of capturing needles facing each other in a three-dimensional space; (b) means for applying an electric field between the pair of capturing needles to capture biological substances; (c) A biologically related substance capturing device characterized in that the biologically related substance is observed three-dimensionally.
ル電極を形成してなる請求項７記載の生体関連物質の捕
捉装置。8. The biologically related substance trapping device according to claim 7, wherein a radial electrode is formed around the biologically related substance.
界をかけ、生体関連物質を捕捉し、該生体関連物質を探
針により三次元的に観察する生体関連物質の観察方法。9. A method for observing a biologically related substance, in which an electric field is applied between a pair of trapping needles facing each other in space, a biologically related substance is captured, and the biologically related substance is observed three-dimensionally with a probe.
電界をかけ、生体関連物質を捕捉し、前記生体関連物質
を中心としたラジアル電極を設け、該ラジアル電極への
電位の印加により、生体関連物質を回転させながら探針
により三次元的に観察する生体関連物質の観察方法。10. Applying an electric field between a pair of capturing needles facing each other in space to capture a biological substance, providing a radial electrode centered on the biological substance, and applying a potential to the radial electrode, A method for observing biological substances in three dimensions using a probe while rotating the biological substances.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP859891A JP2937498B2 (en) | 1991-01-28 | 1991-01-28 | Apparatus for capturing biological substances and method for observing biological substances using the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP859891A JP2937498B2 (en) | 1991-01-28 | 1991-01-28 | Apparatus for capturing biological substances and method for observing biological substances using the same |
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| Publication Number | Publication Date |
|---|---|
| JPH04250332A true JPH04250332A (en) | 1992-09-07 |
| JP2937498B2 JP2937498B2 (en) | 1999-08-23 |
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ID=11697408
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005501251A (en) * | 2001-08-24 | 2005-01-13 | アプレラ コーポレイション | Working with analytes using electric fields |
-
1991
- 1991-01-28 JP JP859891A patent/JP2937498B2/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005501251A (en) * | 2001-08-24 | 2005-01-13 | アプレラ コーポレイション | Working with analytes using electric fields |
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|---|---|
| JP2937498B2 (en) | 1999-08-23 |
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