JP2002357534A - Method for measuring particle - Google Patents
Method for measuring particleInfo
- Publication number
- JP2002357534A JP2002357534A JP2002078547A JP2002078547A JP2002357534A JP 2002357534 A JP2002357534 A JP 2002357534A JP 2002078547 A JP2002078547 A JP 2002078547A JP 2002078547 A JP2002078547 A JP 2002078547A JP 2002357534 A JP2002357534 A JP 2002357534A
- Authority
- JP
- Japan
- Prior art keywords
- particles
- particle
- image
- opaque
- suspension
- 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.)
- Pending
Links
- 239000002245 particle Substances 0.000 title claims abstract description 201
- 238000000034 method Methods 0.000 title claims abstract description 19
- 239000000725 suspension Substances 0.000 claims abstract description 78
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- 239000010419 fine particle Substances 0.000 claims abstract description 23
- 238000003384 imaging method Methods 0.000 claims abstract description 7
- 238000002156 mixing Methods 0.000 claims abstract description 5
- 239000011521 glass Substances 0.000 claims description 26
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 23
- 229910052799 carbon Inorganic materials 0.000 claims description 23
- 230000001678 irradiating effect Effects 0.000 claims description 9
- 238000005259 measurement Methods 0.000 claims description 7
- 239000011859 microparticle Substances 0.000 claims description 3
- 238000000691 measurement method Methods 0.000 claims 1
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- 238000009826 distribution Methods 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- 238000012545 processing Methods 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 5
- 239000008213 purified water Substances 0.000 description 5
- 238000010191 image analysis Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 239000004925 Acrylic resin Substances 0.000 description 3
- 229920000178 Acrylic resin Polymers 0.000 description 3
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 3
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000010445 mica Substances 0.000 description 3
- 229910052618 mica group Inorganic materials 0.000 description 3
- 238000001000 micrograph Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- OGIDPMRJRNCKJF-UHFFFAOYSA-N titanium oxide Inorganic materials [Ti]=O OGIDPMRJRNCKJF-UHFFFAOYSA-N 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 238000003703 image analysis method Methods 0.000 description 2
- 229910052809 inorganic oxide Inorganic materials 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 229910004298 SiO 2 Inorganic materials 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 239000011362 coarse particle Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 230000005588 protonation Effects 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
- G01N15/147—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle the analysis being performed on a sample stream
Landscapes
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Image Analysis (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、粒子測定方法に関
し、特に透明粒子を撮像して画像解析する技術に関す
る。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for measuring particles, and more particularly, to a technique for imaging transparent particles and analyzing the images.
【０００２】[0002]
【従来の技術】塗料や化粧品等に混入される透明粒子や
透明フレーク（flake）の品質を管理する上で、それら
の形状やサイズを測定して管理することは非常に重要で
ある。従来の粒子画像解析法においては、粒子像の背景
に対する輝度差を用いて、ある閾値で粒子像をデジタル
化（２値化）し、得られた２値化画像を解析する（例え
ば、特開平８−１３６４３９号公報参照）。2. Description of the Related Art In controlling the quality of transparent particles and flakes mixed in paints and cosmetics, it is very important to measure and control their shapes and sizes. In a conventional particle image analysis method, a particle image is digitized (binarized) at a certain threshold value using a luminance difference of the particle image with respect to the background, and the obtained binarized image is analyzed (for example, 8-136439).
【０００３】[0003]
【発明が解決しようとする課題】ここで、図１を用い
て、不透明粒子の粒子画像と透明粒子の粒子画像につい
て説明する。図１の左側の不透明粒子の粒子画像Ａは、
２値化閾値Ｔｈに対して粒子輝度信号ＳＡが十分に大き
い。そのため２値化後の粒子信号ＰＡは実際の粒子の大
きさを正確に反映した値となっている。それに対して、
図１の右側の透明粒子の粒子画像Ｂは、２値化閾値Ｔｈ
に対して粒子輝度信号ＳＢが非常に小さい。そのため、
２値化後の粒子信号ＰＢは実際の粒子の大きさを反映し
た値とならず、２つの小さい粒子として捉えられてしま
う。すなわち、粒子と背景の輝度差が小さい透明性が高
い粒子の場合、正確な画像解析を行うことは非常に困難
である。Here, a particle image of opaque particles and a particle image of transparent particles will be described with reference to FIG. The particle image A of the opaque particles on the left side of FIG.
The particle luminance signal SA is sufficiently large with respect to the binarization threshold Th. Therefore, the particle signal PA after binarization has a value that accurately reflects the actual particle size. On the other hand,
The particle image B of the transparent particles on the right side of FIG.
The particle brightness signal SB is very small. for that reason,
The particle signal PB after binarization does not have a value reflecting the actual size of the particle, and is regarded as two small particles. That is, in the case of a highly transparent particle having a small difference in brightness between the particle and the background, it is very difficult to perform accurate image analysis.
【０００４】従来のフロー式粒子分析装置を用いて、ガ
ラス粒子を測定した場合、単一粒子像の比較的輝度差の
大きい粒子輪郭の一部、或いは粒子表面の凹凸の一部を
別々の粒子像と誤認識し、それらを画像解析してしまう
結果、以下の不具合が発生する。 １）単一粒子像の一部を１つの粒子像として画像処理す
る。つまり、図2に示すように透明粒子の画像のほとん
どが、線状画像として認識される。これが、図4に示さ
れるような不透明粒子の画像であれば、正しく認識され
るはずである。 ２）誤認識された粒子像は円形度が極端に低値となる。
つまり、そのほとんどが線状画像として認識される結
果、円形度＝（粒子像と同じ投影面積値を持つ円の周囲
長）／（粒子投影像の周囲長）が低くなる。当然、これ
らの誤認識された粒子像から得られた解析結果は誤った
値となる。When glass particles are measured using a conventional flow-type particle analyzer, a part of a particle contour having a relatively large difference in luminance of a single particle image or a part of irregularities on a particle surface is separated into separate particles. As a result of erroneously recognizing the images and analyzing the images, the following problems occur. 1) Image processing is performed on a part of a single particle image as one particle image. That is, as shown in FIG. 2, most of the images of the transparent particles are recognized as linear images. If this is an image of opaque particles as shown in FIG. 4, it should be correctly recognized. 2) The particle degree of erroneous recognition has an extremely low circularity.
That is, as a result, most of them are recognized as linear images, the circularity = (perimeter of a circle having the same projected area value as the particle image) / (perimeter of the particle projection image) is reduced. Of course, the analysis results obtained from these misrecognized particle images will have incorrect values.
【０００５】本発明は上記事情に鑑みてなされたもの
で、透明粒子に不透明微粒子を付着させることにより、
画像処理法において背景との十分な輝度差を確保し、正
確な画像解析を可能とする粒子測定方法を提供するもの
である。[0005] The present invention has been made in view of the above circumstances, and by attaching opaque fine particles to transparent particles,
It is an object of the present invention to provide a particle measuring method which ensures a sufficient luminance difference from a background in an image processing method and enables accurate image analysis.
【０００６】[0006]
【課題を解決するための手段】本発明は、水に接触する
と第１表面電位を生じる透明粒子を水に分散させ第１懸
濁液を作成する工程と、水に接触すると第１表面電位と
極性の異なる第２表面電位を生じる不透明微粒子を水に
分散させ第２懸濁液を作成する工程と、不透明微粒子が
透明粒子に静電的に付着して透明粒子を不透明化するよ
うに第１および第２懸濁液を混合する工程と、不透明化
粒子に光を照射して撮像する工程と、撮像された画像を
解析する工程を含む粒子測定方法を提供するものであ
る。According to the present invention, there is provided a process for preparing a first suspension by dispersing transparent particles, which generate a first surface potential when contacted with water, in water; Forming a second suspension by dispersing opaque microparticles having a second surface potential having different polarities in water; And a step of mixing the second suspension, irradiating the opacified particles with light to capture an image, and analyzing the captured image.
【０００７】[0007]
【発明の実施の形態】この発明による粒子測定方法は、
水に接触すると第１表面電位を生じる透明粒子を水に分
散させ第１懸濁液を作成する工程と、水に接触すると第
１表面電位と極性の異なる第２表面電位を生じる不透明
微粒子を水に分散させ第２懸濁液を作成する工程と、不
透明微粒子が透明粒子に静電的に付着して透明粒子を不
透明化するように第１および第２懸濁液を混合する工程
と、不透明化粒子に光を照射して撮像する工程と、撮像
された画像を解析する工程を含むことを特徴とする。こ
の発明の測定対象の透明粒子としては、ガラス、アクリ
ル樹脂又はマイカからなる粒子やフレークが挙げられ
る。不透明微粒子としては、酸化チタンやカーボンから
なる微粒子が挙げられる。この発明における粒子、微粒
子への帯電は、次のような現象Ａ、Ｂ（北原文雄他著
「ゼータ電位」第94頁、1995年１月、サイエンティスト
社、参照）に着目されてなされたものである。 Ａ．ガラス、酸化チタン、マイカのような無機酸化物の
粒子は、水と接すると水和を起こして、必ずＯＨ基を有
する。つまり、その水のｐＨ値によって粒子の表面電位
が変化する。例えば、ＳｉＯ2粒子を例にとると次のよ
うに表される。 ＝Ｓｉ・ＯＨ＋Ｈ+⇒＝Ｓｉ・ＯＨ2 +＋ＯＨ- ＝Ｓｉ・ＯＨ＋ＯＨ-⇒＝Ｓｉ・Ｏ-＋Ｈ2Ｏ+Ｈ+ 従って、低いｐＨではプロトン付加により正の電位を帯
び、ｐＨが高くなるとＯＨ基からのプロトンの引き抜き
で負に帯電する。 Ｂ．アクリル樹脂やカーボンからなる粒子は、粒子表面
にフェノール性ＯＨ基やＣＯＯＨ基などの酸性基などを
もつので、水中では負に帯電するが、ｐＨによって表面
電位の極性を反転させることはできない。従って、この
発明において、第１懸濁液の中の透明粒子がガラスやマ
イカのような無機酸化物からなる場合には、それらの表
面電位は現象Ａに基づいて正電位をもつ。このとき、第
２懸濁液中の不透明微粒子としてカーボン粒子を用いる
と、現象Ｂに基づいて、表面電位を負電位にすることが
できる。また、第１懸濁液中の透明粒子がアクリル樹脂
からなる場合は、その表面電位は、現象Ｂに基づいて、
負電位となる。この場合には、第２懸濁液中の不透明微
粒子に酸化チタン粒子を用いると現象Ａに基づいてその
表面電位を正電位にすることができる。なお、第１およ
び第２懸濁液に用いる水としては、精製水を挙げること
ができる。必要であれば、原理Ａに基づいて、第１また
は第２懸濁液のｐＨを変化させて表面電位を制御するこ
とができる。また、第１又は第２懸濁液のｐＨの調整
は、予め水のｐＨを調整することにより行ってもよい。
水又は懸濁液のｐＨの調整には、水酸化カリウム、水酸
化ナトリウム、アンモニア、酢酸、塩酸、硝酸等の酸や
塩基を用いることができるが、粒子又は微粒子と反応し
ないものを用いることが肝要である。本発明における粒
子および微粒子の表面電位とは、それらが、水中に十分
に分散している状態にあるときの表面電位を意味する。
その電位は、ゼータ（Zeta）電位測定において、透明粒
子と不透明粒子とは、互いに極性の異なる３０ｍＶ以上
の表面電位を有することが好ましい。なお、ゼータ電位
の測定は、電気泳動法を用いた市販のゼータ電位計、例
えばZETASIZER（Malvern Instruments Ltd.製）を用い
ることができる。BEST MODE FOR CARRYING OUT THE INVENTION
A step of preparing a first suspension by dispersing, in water, transparent particles that generate a first surface potential when in contact with water; Forming a second suspension by dispersing the first and second suspensions such that the opaque fine particles electrostatically adhere to the transparent particles to make the transparent particles opaque; A step of irradiating the activated particles with light to capture an image, and a step of analyzing the captured image. Examples of the transparent particles to be measured in the present invention include particles and flakes made of glass, acrylic resin, or mica. Opaque fine particles include fine particles made of titanium oxide or carbon. The electrification of the particles and fine particles in the present invention is made by paying attention to the following phenomena A and B (see "Zeta potential", page 94, Jan. 1995, Scientist, etc.) by Fumio Kitahara et al. is there. A. Particles of inorganic oxides such as glass, titanium oxide, and mica hydrate upon contact with water and always have OH groups. That is, the surface potential of the particles changes depending on the pH value of the water. For example, taking the SiO 2 particles as an example, they are expressed as follows. = Si.OH + H + ⇒ = Si.OH 2 + + OH − = Si.OH + OH − ⇒ = Si.O − + H 2 O + H + Therefore, at a low pH, a positive potential is obtained by protonation, and when the pH becomes high, OH It becomes negatively charged by the extraction of a proton from the group. B. Particles made of an acrylic resin or carbon have an acidic group such as a phenolic OH group or a COOH group on the particle surface, and thus are negatively charged in water. However, the polarity of the surface potential cannot be reversed by pH. Therefore, in the present invention, when the transparent particles in the first suspension are made of an inorganic oxide such as glass or mica, their surface potential has a positive potential based on the phenomenon A. At this time, if carbon particles are used as the opaque fine particles in the second suspension, the surface potential can be made negative based on the phenomenon B. Further, when the transparent particles in the first suspension are made of an acrylic resin, the surface potential thereof is based on the phenomenon B,
The potential becomes negative. In this case, if titanium oxide particles are used as the opaque fine particles in the second suspension, the surface potential can be made positive based on the phenomenon A. In addition, purified water can be mentioned as water used for the first and second suspensions. If necessary, based on principle A, the surface potential can be controlled by changing the pH of the first or second suspension. Adjustment of the pH of the first or second suspension may be performed by adjusting the pH of water in advance.
To adjust the pH of water or the suspension, an acid or base such as potassium hydroxide, sodium hydroxide, ammonia, acetic acid, hydrochloric acid, or nitric acid can be used, but one that does not react with particles or fine particles can be used. It is important. The surface potential of the particles and fine particles in the present invention means the surface potential when they are sufficiently dispersed in water.
As for the potential, in the measurement of the zeta potential, it is preferable that the transparent particles and the opaque particles have a surface potential of 30 mV or more having different polarities from each other. The zeta potential can be measured using a commercially available zeta potentiometer using electrophoresis, for example, ZETASIZER (manufactured by Malvern Instruments Ltd.).
【０００８】この発明の測定対象の透明粒子は中位径
（mean diameter）で１〜１００μｍであることが好ま
しい。The transparent particles to be measured in the present invention preferably have a mean diameter of 1 to 100 μm.
【０００９】不透明微粒子は、撮像装置の分解能より小
さいことが必要であるため、直径が中位径で５０〜２０
０ｎｍが好ましい。つまり、不透明微粒子が透明粒子に
対して十分に小さければ、画像解析時に不透明微粒子が
透明粒子の大きさ・形状に及ぼす影響が他の誤差要因に
対して十分無視できる。なお、不透明粒子の懸濁液は、
カーボンインクを用いて調製されてもよい。Since the opaque fine particles need to be smaller than the resolution of the image pickup device, they have a median diameter of 50 to 20.
0 nm is preferred. That is, if the opaque fine particles are sufficiently smaller than the transparent particles, the influence of the opaque fine particles on the size and shape of the transparent particles during image analysis can be sufficiently ignored with respect to other error factors. The suspension of opaque particles is
It may be prepared using carbon ink.
【００１０】光を照射する工程おいては、パルス発光す
るストロボやレーザ光源などを利用できる。In the step of irradiating light, a strobe or a laser light source which emits a pulse can be used.
【００１１】撮像する工程おいては、一般的な２次元画
像を撮像するビデオカメラなどを利用できる。In the image capturing step, a video camera or the like for capturing a general two-dimensional image can be used.
【００１２】解析する工程においては、ＣＰＵ、ＲＯ
Ｍ、ＲＡＭ、およびＩ／Ｏポートからなるマイクロコン
ピュータあるいは市販のパーソナルコンピュータを利用
することができる。画像を解析する工程が、画像を２値
化して画像データとし、画像データに基づいて粒子の円
形度を算出する工程を含んでもよい。In the step of analyzing, CPU, RO
A microcomputer including an M, a RAM, and an I / O port or a commercially available personal computer can be used. The step of analyzing the image may include a step of binarizing the image into image data and calculating a circularity of the particle based on the image data.
【００１３】不透明化粒子に光を照射して撮像する工程
が、不透明化粒子をフローセルに流す工程と、フローセ
ル中を流れる不透明化粒子に光を照射して撮像する工程
を含んでもよい。微粒子付粒子をフローセルに流す工程
においては、粒子懸濁液の流れをシース液で包んで流す
ことにより流体力学的効果によって、細いあるいは偏平
な流れに変換することができるシースフローセルを利用
できる。[0013] The step of irradiating the opacifying particles with light and imaging may include the step of flowing the opacifying particles to the flow cell and the step of irradiating light to the opacifying particles flowing in the flow cell and imaging. In the step of flowing the particles with fine particles into the flow cell, a sheath flow cell that can be converted into a thin or flat flow by a hydrodynamic effect by wrapping the flow of the particle suspension with a sheath liquid and flowing the same can be used.
【００１４】実施例 (1) 測定対象となる透明ガラス粒子懸濁液の調製 平均径（mean diameter）が約７０μｍの透明ガラスフ
レーク０．１０ｍｇを約２５ｍＬの精製水（ｐＨ：６．
５）に分散させ懸濁液を調製する。Example (1) Preparation of suspension of transparent glass particles to be measured 0.10 mg of transparent glass flake having a mean diameter of about 70 μm was added to about 25 mL of purified water (pH: 6.
Disperse in 5) to prepare a suspension.
【００１５】(2) 不透明微粒子懸濁液の調製 市販のカーボンインク（濃度０．２重量％、カーボン粒
子中位径１００ｎｍ）１ｍＬを約５０ｍＬの精製水（ｐ
Ｈ：６．５）で希釈しカーボン粒子懸濁液を調製する。(2) Preparation of opaque fine particle suspension 1 mL of commercially available carbon ink (concentration: 0.2% by weight, median diameter of carbon particles: 100 nm) is added to about 50 mL of purified water (p
H: 6.5) to prepare a carbon particle suspension.
【００１６】(3) 不透明微粒子を付着させた透明ガラス
粒子懸濁液の調製 (1)の透明ガラス粒子懸濁液２５ｍＬと(2)のカーボン粒
子懸濁液５ｍＬを混合する。(3) Preparation of suspension of transparent glass particles to which opaque fine particles are adhered 25 mL of the suspension of transparent glass particles of (1) and 5 mL of the suspension of carbon particles of (2) are mixed.
【００１７】(4) ゼータ電位の測定 (1)、(2)、(3)の懸濁液をゼータ電位計（マルバーン社
製）で測定する。(1)で得られた粒子懸濁液と、(2)で得
られた粒子懸濁液のゼータ電位は、＋４１．６ｍＶ、−
３７．５ｍＶとなる。＋４１．６ｍＶのゼータ電位をも
つ透明ガラス粒子懸濁液(1)と、−３７．５ｍＶのゼー
タ電位をもつカーボン粒子懸濁液(2)を混合した結果、
混合試料(3)のゼータ電位は−３３．２ｍＶとなりほぼ
カーボン粒子懸濁液のそれと同程度になる。つまり、こ
れは両者の表面電位の差によって透明ガラス粒子表面に
カーボン粒子が吸着され、それによって、透明ガラス粒
子表面の見かけ上の電位がカーボン粒子のそれと置き換
わる結果である。(4) Measurement of zeta potential The suspensions of (1), (2) and (3) are measured with a zeta potential meter (Malvern). The zeta potential of the particle suspension obtained in (1) and the particle suspension obtained in (2) are +41.6 mV, −
It becomes 37.5 mV. As a result of mixing a transparent glass particle suspension (1) having a zeta potential of +41.6 mV and a carbon particle suspension (2) having a zeta potential of -37.5 mV,
The zeta potential of the mixed sample (3) is -33.2 mV, which is almost the same as that of the carbon particle suspension. In other words, this is a result of the carbon particles being adsorbed on the surface of the transparent glass particles due to the difference in surface potential between the two, thereby replacing the apparent potential of the surface of the transparent glass particles with that of the carbon particles.
【００１８】(5) フロー式粒子画像分析装置による測定 (1)の粒子懸濁液、(3)の粒子懸濁液をそれぞれフロー式
粒子分析装置で測定し、得られた撮像画像を図２、図４
に、円形度分布を図５、図７に示す。(1)の透明ガラス
粒子懸濁液のみを測定した場合の撮像画像は、図２に示
したように、粒子像と背景の輝度差が小さい。そのため
に粒子像の一部が１つの線状粒子として誤認識される。
つまり、図５に示したように、解析結果の円形度分布に
おいて円形度が下限値０．４に偏って集中し、誤った粒
子像を画像解析していることが明らかである。それに対
して、(3)の混合懸濁液の場合、撮像画像は図4に示した
ように、粒子像と背景の輝度差が大きい。そのために１
つの粒子像の全体がひとつの粒子として認識される。つ
まり、図７に示したように、円形度分布において、円形
度が実際の粒子形状に相応した領域に分布しており、粒
子画像を正確に画像解析していることがわかる。(5) Measurement by flow type particle image analyzer The particle suspension of (1) and the particle suspension of (3) were each measured by a flow type particle analyzer, and the obtained captured image was shown in FIG. , FIG.
5 and 7 show the circularity distribution. In the captured image obtained by measuring only the transparent glass particle suspension of (1), as shown in FIG. 2, the difference in luminance between the particle image and the background is small. Therefore, a part of the particle image is erroneously recognized as one linear particle.
That is, as shown in FIG. 5, it is apparent that the circularity is concentrated to the lower limit value 0.4 in the circularity distribution of the analysis result, and that an erroneous particle image is image-analyzed. In contrast, in the case of the mixed suspension (3), the captured image has a large difference in luminance between the particle image and the background, as shown in FIG. 1 for that
The whole of one particle image is recognized as one particle. That is, as shown in FIG. 7, in the circularity distribution, the circularity is distributed in a region corresponding to the actual particle shape, and it can be seen that the particle image is accurately analyzed.
【００１９】比較例 ここまでは、逆極性のゼータ電位を持った粒子懸濁液の
混合によって、粒子の表面電位による付着現象について
説明したが、次に比較例として、同じ粒子を用いて粒子
懸濁液のゼータ電位を同極性に調整した場合について実
験を試みる。Comparative Example Up to this point, the adhesion phenomenon due to the surface potential of particles by mixing particle suspensions having zeta potentials of opposite polarities has been described. Next, as a comparative example, the same particles were used to suspend particles. Experiments are performed on the case where the zeta potential of the suspension is adjusted to the same polarity.
【００２０】(6) ｐＨ１０の透明ガラス粒子懸濁液の調
製 平均径が約７０μｍの透明ガラスフレーク０．１０ｍｇ
を約２５ｍＬの精製水に分散させ、０．１モルのＮａＯ
ＨでｐＨ１０に調整して粒子懸濁液を調製する。透明ガ
ラス粒子のゼータ電位は、ＮａＯＨの添加によるｐＨ
（potential of hydrogen）調整によって、＋符号から
−符号へ極性が変化する。(6) Preparation of a suspension of transparent glass particles having a pH of 10 0.10 mg of transparent glass flakes having an average diameter of about 70 μm
Is dispersed in about 25 mL of purified water, and 0.1 M NaO
Adjust the pH to 10 with H to prepare a particle suspension. The zeta potential of the transparent glass particles is determined by the pH due to the addition of NaOH.
By adjusting (potential of hydrogen), the polarity changes from the plus sign to the minus sign.
【００２１】(7) ｐＨ１０の不透明微粒子懸濁液の調製 市販のカーボンインク１ｍＬを５０ｍＬの精製水で希釈
し、０．１モルのＮａＯＨでｐＨ１０に調整してカーボ
ン粒子懸濁液試料を調製する。カーボン粒子懸濁液のゼ
ータ電位は、ＮａＯＨの添加によるｐＨ調整を行った場
合にも極性変化は生じない。(7) Preparation of opaque fine particle suspension of pH 10 1 mL of commercially available carbon ink is diluted with 50 mL of purified water and adjusted to pH 10 with 0.1 M NaOH to prepare a carbon particle suspension sample. . The polarity of the zeta potential of the carbon particle suspension does not change even when the pH is adjusted by adding NaOH.
【００２２】(8) ｐＨ１０の透明ガラス粒子懸濁液とｐ
Ｈ１０のカーボンの粒子懸濁液の混合物の調製する。 (6)の透明ガラス粒子の粒子懸濁液２５ｍＬと(7)のカー
ボン粒子懸濁液５ｍＬを混合する。(8) pH 10 transparent glass particle suspension and p
Prepare a mixture of H10 carbon particle suspension. 25 mL of the transparent glass particle suspension (6) and 5 mL of the carbon particle suspension (7) are mixed.
【００２３】(9) ゼータ電位の測定 (6)、(7)、(8)の粒子懸濁液をゼータ電位計でそれぞれ
測定する。(6)の粒子懸濁液のゼータ電位は−４０．８
ｍＶ、(7)の粒子懸濁液のゼータ電位は−２７．０ｍ
Ｖ、(8)の粒子懸濁液のゼータ電位は−２８．１ｍＶと
なる。(8)の粒子懸濁液のゼータ電位は、(7)の粒子懸濁
液のゼータ電位に置き換わるように見えるが、これは
(7)の粒子懸濁液の粒子数が圧倒的に多いためである。(9) Measurement of zeta potential The particle suspensions of (6), (7) and (8) are each measured with a zeta potential meter. The zeta potential of the particle suspension of (6) is -40.8
mV, the zeta potential of the particle suspension of (7) is -27.0 m
V, the zeta potential of the particle suspension of (8) is -28.1 mV. The zeta potential of the particle suspension of (8) appears to be replaced by the zeta potential of the particle suspension of (7).
This is because the number of particles in the particle suspension of (7) is overwhelmingly large.
【００２４】(10) フロー式粒子画像分析装置による測
定 (8)の粒子懸濁液をフロー式粒子分析装置で測定し、得
られた撮像画像を図３に、円形度分布を図６に示す。図
３に示したように、(8)の粒子懸濁液においては、ガラ
ス粒子へのカーボン粒子の表面電荷による吸着が発生せ
ず、測定結果はカーボン粒子を混入させない場合と同様
に、画像処理において誤認識が生じる。つまり、(8)の
粒子懸濁液は円形度が図６に示したように、下限値０．
４以下に集中し、粒子像の誤認識によって、粒子像の一
部を線状画像として画像解析していることがわかる。(10) Measurement by Flow Type Particle Image Analyzer The particle suspension of (8) is measured by a flow type particle analyzer, and the obtained captured image is shown in FIG. 3 and the circularity distribution is shown in FIG. . As shown in FIG. 3, in the particle suspension of (8), the adsorption of the carbon particles to the glass particles due to the surface charge did not occur, and the measurement results showed that the image processing was the same as in the case where the carbon particles were not mixed. Causes misrecognition. In other words, the particle suspension of (8) has a circularity of lower limit of 0. 0, as shown in FIG.
It is understood that a part of the particle image is image-analyzed as a linear image due to erroneous recognition of the particle image.
【００２５】本発明の粒子測定法によれば、透明なガラ
ス粒子に逆表面電位の不透明微粒子を付着させることに
より、撮像画像における背景と粒子像との間に十分な輝
度差が確保でき、粒子の正確な画像解析が行える。According to the particle measuring method of the present invention, by attaching opaque fine particles having an opposite surface potential to transparent glass particles, a sufficient luminance difference between the background and the particle image in the captured image can be secured. Can perform accurate image analysis.
【００２６】フロー式粒子画像分析装置の構成と動作 ここで、前述のフロー式粒子画像分析装置について説明
する。この装置では、図８に示すように、まず粒子懸濁
液はダイヤフラムポンプ等の吸引手段（図示していな
い）によって吸引ピペット１から吸引され、サンプルフ
ィルター２を通りフローセル５の上部の試料チャージン
グライン３へ引き込まれる。サンプルフィルター２によ
って、懸濁液中の粗大な粒子やごみが取り除かれ、流路
の細い（狭い）フローセル５が詰まらないようにしてい
る。Structure and operation of the flow type particle image analyzer The flow type particle image analyzer described above will now be described. In this apparatus, first, as shown in FIG. 8, a particle suspension is sucked from a suction pipette 1 by suction means (not shown) such as a diaphragm pump, and then passed through a sample filter 2 to charge a sample on an upper part of a flow cell 5. Pulled into line 3. The sample filter 2 removes coarse particles and dust in the suspension, and prevents the flow cell 5 having a narrow (narrow) flow path from being clogged.
【００２７】チャージングライン３に引き込まれた粒子
懸濁液は、シースシリンジ４を動作させることによって
フローセル５に導かれ、図９に示すようにサンプルノズ
ル５ａの先端から懸濁液が少しずつ押し出される。それ
と同時にシース液ＳＬもシース液ボトル６からシース液
チャンバー７を介してフローセル５に送り込まれ、粒子
懸濁液はそのシース液ＳＬで取り囲まれ、図９に示すよ
うに、液体力学的に懸濁液流は偏平に絞られてフローセ
ル５の内を流れ、廃液チャンバー１４を介して排出され
る。このように偏平に絞られた懸濁液流に対して、スト
ロボ８からパルス光を１／３０秒ごとに周期的に照射す
ることによって、１／３０秒ごとに粒子の静止画像が対
物レンズ９を介してビデオカメラ１０で撮像される。The particle suspension drawn into the charging line 3 is guided to the flow cell 5 by operating the sheath syringe 4, and the suspension is gradually extruded from the tip of the sample nozzle 5a as shown in FIG. It is. At the same time, the sheath liquid SL is also sent from the sheath liquid bottle 6 to the flow cell 5 via the sheath liquid chamber 7, and the particle suspension is surrounded by the sheath liquid SL and hydrodynamically suspended as shown in FIG. The liquid flow is flattened, flows through the flow cell 5, and is discharged through the waste liquid chamber 14. By irradiating the flattened suspension flow with pulse light periodically from the strobe 8 every 1/30 second, a still image of particles is obtained every 1/30 second. Is captured by the video camera 10 via the.
【００２８】ビデオカメラ１０からの画像信号は、画像
処理装置１１で処理され、モニターテレビ１２に表示さ
れる。１３は各種の操作等を行うためのキーボードであ
る。An image signal from the video camera 10 is processed by the image processing device 11 and displayed on the monitor television 12. Reference numeral 13 denotes a keyboard for performing various operations and the like.
【００２９】そして、画像処理装置１１では、次のよう
にして画像処理が行われる。画像信号は、画像処理装置
１１に取り込まれて画像データとしてＡ／Ｄ変換され
る。次に、画像データをある適当なスレシホールドレベ
ルで２値化し、画像メモリに格納する。Then, the image processing device 11 performs image processing as follows. The image signal is taken into the image processing device 11 and A / D converted as image data. Next, the image data is binarized at an appropriate threshold level and stored in an image memory.
【００３０】次に、図１０のように２値化された粒子像
に対してエッジ点かどうかを判定するとともに、着目し
ているエッジ点に対して隣合うエッジ点がどの方向にあ
るかの情報、すなわちチェインコードを生成する。次
に、このチェインコードを参照しながら図１０に示すよ
うに粒子像のエッジトレースを行い、各粒子像の総画素
数、総エッジ数、斜めエッジ数を求める。Next, it is determined whether or not the binarized particle image is an edge point as shown in FIG. 10, and the direction of the edge point adjacent to the focused edge point is determined. Generate information, ie, chain code. Next, referring to the chain code, an edge trace of the particle image is performed as shown in FIG. 10, and the total number of pixels, the total number of edges, and the number of oblique edges of each particle image are obtained.
【００３１】各粒子像に対して求められた総画素数、総
エッジ数、斜めエッジ数から、下記の式によって各粒子
像の投影面積Ｓと周囲長Ｌを求める。From the total number of pixels, the total number of edges, and the number of oblique edges obtained for each particle image, the projection area S and the perimeter L of each particle image are obtained by the following equation.
【００３２】図１０に示すように、２値画像の周囲のエ
ッジの中心を結んでできる枠内の面積Ｓおよび枠の長さ
（周期長Ｌ）は、１画素当たりの面積を１とした場合、 面積Ｓ＝総画素数−（総エッジ数×０．５）−１……(1) 周囲長Ｌ＝（総エッジ数−斜めエッジ数）＋（斜めエッジ数×√２）……(2)As shown in FIG. 10, the area S in the frame formed by connecting the centers of the edges around the binary image and the length of the frame (period length L) are assuming that the area per pixel is 1. Area S = Total number of pixels− (Total number of edges × 0.5) −1 (1) Perimeter L = (Total number of edges−Number of oblique edges) + (Number of oblique edges × √2) (2) )
【００３３】次に、上記面積Ｓと周囲長Ｌを用いて円相
当径と円形度を求める。円相当径とは、粒子像の投影面
積と同じ面積を持つ円を想定し、その円の直径のことで
あり、式(3)で表される。円形度とは、例えば式(4)で定
義される値であり、粒子像が円形の時に円形度は１にな
り、粒子像が細長くなればなるほど円形度は小さい値に
なる。Next, using the area S and the perimeter L, the equivalent circle diameter and circularity are determined. The equivalent circle diameter is a diameter of the circle assuming a circle having the same area as the projection area of the particle image, and is represented by Expression (3). The circularity is, for example, a value defined by Expression (4). When the particle image is circular, the circularity is 1, and the smaller the particle image is, the smaller the circularity is.
【００３４】 円相当径＝（粒子投影像面積値／π）1/2×２……(3) 円形度＝（粒子像と同じ投影面積値を持つ円の周囲長）／粒子投影像の周囲長 ……(4)Circle equivalent diameter = (particle projected image area value / π) 1/2 × 2 (3) circularity = (perimeter of circle having the same projected area value as particle image) / periphery of particle projected image Long ...... (4)
【００３５】各粒子像の円相当径や円形度が求められれ
ば、次にその値をもとにして粒度分布や円形度分布を作
成し、モニタテレビ１２へ出力する。なお、上記フロー
式粒子画像分析装置には、シスメックス社製ＦＰＩＡ−
２１００を用いることができる。When the equivalent circle diameter and circularity of each particle image are obtained, a particle size distribution and circularity distribution are created based on the values and output to the monitor television 12. The above-mentioned flow type particle image analyzer includes FPIA- manufactured by Sysmex Corporation.
2100 can be used.
【００３６】[0036]
【発明の効果】この発明によれば、透明粒子に不透明微
粒子を付着して不透明化し撮像するので、撮像された粒
子画像と背景との輝度差が大きくなり、透明粒子の形態
に関する解析を精度よく行うことができる。According to the present invention, since the opaque fine particles are attached to the transparent particles to make the image opaque and the image is taken, the brightness difference between the image of the captured particle and the background is increased, and the analysis on the morphology of the transparent particle can be accurately performed. It can be carried out.
【図１】不透明粒子画像と透明粒子画像の２値化の説明
図である。FIG. 1 is an explanatory diagram of binarization of an opaque particle image and a transparent particle image.
【図２】従来の方法によるガラス粒子懸濁液中の粒子の
撮像画像（顕微鏡写真）である。FIG. 2 is a photographed image (micrograph) of particles in a glass particle suspension according to a conventional method.
【図３】ｐＨ１０における鱗片状ガラス粒子懸濁液とカ
ーボンインク粒子懸濁液の混合物中の粒子の撮像画像
（顕微鏡写真）である。FIG. 3 is a photographed image (micrograph) of particles in a mixture of a flaky glass particle suspension and a carbon ink particle suspension at pH 10.
【図４】本発明方法によるガラス粒子懸濁液中の粒子の
撮像画像（顕微鏡写真）である。FIG. 4 is a photographed image (micrograph) of particles in a glass particle suspension according to the method of the present invention.
【図５】図２に対応するガラス粒子懸濁液中の粒子の円
形度分布である。FIG. 5 is a circularity distribution of the particles in the glass particle suspension corresponding to FIG. 2;
【図６】ｐＨ１０におけるガラス粒子懸濁液とカーボン
粒子懸濁液の混合物中の粒子の円形度分布である。FIG. 6 is a circularity distribution of particles in a mixture of a glass particle suspension and a carbon particle suspension at pH 10.
【図７】本発明方法により得られるガラス粒子懸濁液中
の粒子の円形度分布である。FIG. 7 is a circularity distribution of particles in a glass particle suspension obtained by the method of the present invention.
【図８】この発明の方法に用いるシース式粒子画像解析
装置の構成説明図である。FIG. 8 is an explanatory diagram of a configuration of a sheath-type particle image analyzer used in the method of the present invention.
【図９】図８の要部詳細図である。9 is a detailed view of a main part of FIG.
【図１０】図８に示す装置の画像解析法を説明する説明
図である。FIG. 10 is an explanatory diagram illustrating an image analysis method of the apparatus shown in FIG.
１ 吸引ピペット ２ サンプルフィルター ３ チャージングライン ４ シースシリンジ ５ フローセル ５ａ サンプルノズル ６ シース液ボトル ７ シース液チャンバー ８ ストロボ ９ 対物レンズ １０ ビデオカメラ １１ 画像処理装置 １２ モニターテレビ １３ キーボード DESCRIPTION OF SYMBOLS 1 Suction pipette 2 Sample filter 3 Charging line 4 Sheath syringe 5 Flow cell 5a Sample nozzle 6 Sheath liquid bottle 7 Sheath liquid chamber 8 Strobe 9 Objective lens 10 Video camera 11 Image processing device 12 Monitor television 13 Keyboard
Claims (11)
明粒子を水に分散させ第１懸濁液を作成する工程と、水
に接触すると第１表面電位と極性の異なる第２表面電位
を生じる不透明微粒子を水に分散させ第２懸濁液を作成
する工程と、不透明微粒子が透明粒子に静電的に付着し
て透明粒子を不透明化するように第１および第２懸濁液
を混合する工程と、不透明化粒子に光を照射して撮像す
る工程と、撮像された画像を解析する工程を含む粒子測
定方法。1. a step of preparing a first suspension by dispersing transparent particles that generate a first surface potential when contacted with water in water, and a second surface potential having a polarity different from the first surface potential when contacted with water. Dispersing the resulting opaque microparticles in water to form a second suspension, and mixing the first and second suspensions such that the opaque microparticles electrostatically adhere to the transparent particles to opaque the transparent particles. A particle measuring method, which includes a step of irradiating the opaque particles with light to capture an image, and a step of analyzing the captured image.
１記載の粒子測定方法。2. The method according to claim 1, wherein the transparent particles are glass flakes.
項１記載の粒子測定方法。3. The method according to claim 1, wherein the opaque fine particles are carbon particles.
の極性を有し、第２懸濁液中の不透明粒子の表面電位が
負の極性を有する請求項１記載の粒子測定方法。4. The particle measurement according to claim 1, wherein the surface potential of the transparent particles in the first suspension has a positive polarity, and the surface potential of the opaque particles in the second suspension has a negative polarity. Method.
が３０ｍＶ以上である請求項１記載の粒子測定方法。5. The method according to claim 1, wherein the surface potential of the transparent particles and the opaque fine particles is 30 mV or more.
し、不透明微粒子５０〜２００ｎｍの中位径を有する請
求項１記載の粒子測定方法。6. The method according to claim 1, wherein the transparent particles have a median diameter of 1 to 100 μm and opaque fine particles have a median diameter of 50 to 200 nm.
成される請求項１記載の粒子測定方法。7. The method according to claim 1, wherein the second suspension is formed using carbon ink.
程が、不透明化粒子をフローセルに流す工程と、フロー
セル中を流れる不透明化粒子に光を照射して撮像する工
程からなる請求項１記載の粒子測定方法。8. The step of irradiating the opaque particles with light and imaging the opaque particles includes the steps of flowing the opaque particles through a flow cell and irradiating the opaque particles with light and imaging the flow particles. The particle measurement method described in the above.
カメラを用いて行われる請求項１記載の粒子測定方法。9. The method according to claim 1, wherein the step of imaging the opaque particles is performed using a video camera.
して画像データとし、画像データに基づいて粒子の円形
度を算出する工程を含む請求項１記載の粒子測定方法。10. The particle measuring method according to claim 1, wherein the step of analyzing the image includes a step of binarizing the image into image data and calculating a circularity of the particle based on the image data.
方のｐＨを制御して、第１および第２表面電位の少なく
とも一方を調整する工程をさらに備える請求項１記載の
粒子測定方法。11. The method according to claim 1, further comprising the step of controlling at least one of the first and second suspensions to adjust at least one of the first and second surface potentials.
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| JPS5481848A (en) * | 1977-11-16 | 1979-06-29 | Matsushita Electric Ind Co Ltd | Conductive image forming particles |
| JPS6017454A (en) * | 1983-07-08 | 1985-01-29 | Matsushita Electric Ind Co Ltd | Image forming grains |
| JPH085543A (en) * | 1994-04-21 | 1996-01-12 | Hitachi Ltd | Method for monitoring dyeing liquid for particle analysis and calibration method for particle analysis |
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