JPS6151569A - Cell identifying device - Google Patents
Cell identifying deviceInfo
- Publication number
- JPS6151569A JPS6151569A JP59174505A JP17450584A JPS6151569A JP S6151569 A JPS6151569 A JP S6151569A JP 59174505 A JP59174505 A JP 59174505A JP 17450584 A JP17450584 A JP 17450584A JP S6151569 A JPS6151569 A JP S6151569A
- Authority
- JP
- Japan
- Prior art keywords
- fluorescence
- spectral distribution
- scattered light
- light
- flow
- 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
- 230000003595 spectral effect Effects 0.000 claims abstract description 24
- 230000005284 excitation Effects 0.000 claims description 14
- 239000000835 fiber Substances 0.000 claims description 9
- 230000003287 optical effect Effects 0.000 claims description 6
- 239000013307 optical fiber Substances 0.000 abstract description 6
- 230000007423 decrease Effects 0.000 abstract description 2
- 230000001678 irradiating effect Effects 0.000 abstract 2
- 210000004027 cell Anatomy 0.000 description 44
- 238000005259 measurement Methods 0.000 description 10
- 238000010586 diagram Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000001514 detection method Methods 0.000 description 3
- 239000002253 acid Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000001506 fluorescence spectroscopy Methods 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- UJKPHYRXOLRVJJ-MLSVHJFASA-N CC(O)C1=C(C)/C2=C/C3=N/C(=C\C4=C(CCC(O)=O)C(C)=C(N4)/C=C4\N=C(\C=C\1/N\2)C(C)=C4C(C)O)/C(CCC(O)=O)=C3C Chemical class CC(O)C1=C(C)/C2=C/C3=N/C(=C\C4=C(CCC(O)=O)C(C)=C(N4)/C=C4\N=C(\C=C\1/N\2)C(C)=C4C(C)O)/C(CCC(O)=O)=C3C UJKPHYRXOLRVJJ-MLSVHJFASA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000002902 bimodal effect Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000022534 cell killing Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 210000004881 tumor cell Anatomy 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
- G01N15/1456—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
- G01N15/1459—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals 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)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
Description
【発明の詳細な説明】
［産業上の利用分野］
本発明は、フローサイトメータ等に用いられ、蛍光剤に
よって標識された細胞を識別するだめの細胞識別装置に
関するものである。DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a cell identification device used in a flow cytometer or the like to identify cells labeled with a fluorescent agent.
［従来の技術］
フローサイトメータとは医学分野で近年注目されている
測定装ガであり、高速で流れる細胞浮遊溶液にレーザー
ビーム等を照射し、そのｎｋ乱先による光電信号を検出
し、細胞の性質・構造等を解明する装置である。[Prior art] A flow cytometer is a measurement instrument that has recently attracted attention in the medical field.It irradiates a rapidly flowing cell suspension solution with a laser beam, etc., detects the photoelectric signal from the NK scattering end, and detects cells. This is a device that elucidates the properties and structure of
従来の細胞識別装置は流通部を通過しシース液に包まれ
た細胞にレーザービームを照射し、その結果生ずる前方
及び側方散乱光により、細胞の形状・大きさ・屈折率等
の粒子的性質を１１）ることができる、また、１゛１１
°光剤により染色され得る細胞に対しては、照射光と直
角な方向の側方散乱光から細胞の蛍光を検出することに
より、細胞解析のための重要な情報を求めることができ
る。Conventional cell identification devices irradiate laser beams onto cells that pass through a flow section and are wrapped in sheath liquid, and the resulting forward and side scattered light is used to determine particle properties such as cell shape, size, and refractive index. 11) can be done, and 1゛11
For cells that can be stained with photoreagents, important information for cell analysis can be obtained by detecting cell fluorescence from side-scattered light in a direction perpendicular to the irradiated light.
一般的に、細胞から放射される蛍光をグイロイツクミラ
ーにより赤色と緑色の蛍光に分解し、緑色蛍光強度でデ
オキシリポ核酸（Ｄ Ｎ Ａ）量を、赤色蛍光強度でリ
ポ核酸（ＲＮＡ）量、タンパク質量を計測する。このよ
うに、従来の装置〆Ｌはキ１１°光を赤／緑（又は赤／
青）の２色によって区別する２色法で測定し、それらの
強度及びその比、或いはこれらと前記散乱光による粒子
的性質とを組み合わせて、細胞を識別するための情報と
している。Generally, the fluorescence emitted from cells is separated into red and green fluorescence using a gray mirror, and the green fluorescence intensity is used to determine the amount of deoxyliponucleic acid (DNA), the red fluorescence intensity is used to determine the amount of liponucleic acid (RNA), Measure the amount of protein. In this way, the conventional device 〆L converts the 11° light into red/green (or red/green).
It is measured using a two-color method that distinguishes cells by two colors (blue), and their intensities and their ratios, or a combination of these and the particle properties of the scattered light, are used as information for identifying cells.
ところで、このような測定を行う場合に、流れている細
胞に入射するレーザービームの大きさは、細胞と同程度
か或いは少し太き目にしている。つまり、細胞の流れに
昨直な方向をＸ方向、流れに平行な方向をＺ方向とする
と、レーザービームの照射域はＸ方向では流通部の壁間
の凹陥よりも小さく、かつ流通部を通過中の細胞のｌＱ
れのゆらぎ、即ち流れの中心軸からのずれによる影響が
無視できる程度の長さ１例えば１００〜２００ ｐｍに
なるようにしている。また、Ｚ方向には通常は細胞の大
きさ程度１例えば１０〜３０７１ｍ、或いはスリットス
キャンニング方式の場合では細胞よりも十分小さい１〜
２ｋＬｍ程度に集光するようにしている。そのために、
第１図の従来の励起光学系の構成図に示すように、２枚
の凸円柱レンズｌ、２を組み合わせて、流通部における
レーザービームの照射域Ｆを調整するようになっている
。By the way, when performing such measurements, the size of the laser beam incident on the flowing cells is set to be about the same size as the cells, or a little thicker. In other words, if the direction perpendicular to the flow of cells is the X direction, and the direction parallel to the flow is the Z direction, the irradiation area of the laser beam is smaller than the depression between the walls of the flow section in the X direction, and it passes through the flow section. lQ of cells in
The length 1, for example, 100 to 200 pm is made such that the influence of fluctuations in the flow, that is, the influence of deviation from the central axis of the flow can be ignored. In addition, in the Z direction, normally the size of the cell is 1, for example, 10 to 3071 m, or in the case of the slit scanning method, the size is 1 to 3,071 m, which is sufficiently smaller than the cell.
The light is focused to about 2 kLm. for that,
As shown in the configuration diagram of a conventional excitation optical system in FIG. 1, two convex cylindrical lenses 1 and 2 are combined to adjust the irradiation area F of the laser beam in the flow section.
このような従来の２色法では、検出する蛍光の波長帯が
限られるために、対象とする細胞に適した蛍光剤が限定
される上に、多種類の細胞が混在する検出に対してはそ
の分解能が低下する。また、最近注目を集めている癌細
胞に選択的に集まるヘマトポルフィリン誘導体（ＨＰＤ
）による腫瘍細胞の識別では、対象細胞によって２峰性
又は３ＩＩ＋￥性を示すので正確な検出が不可能となる
。更に、強い励起光には殺細胞効果があるため、励起光
強度には限界があって強い蛍光が得られないことなど幾
つかの欠点がある。In such conventional two-color methods, the wavelength band of fluorescence to be detected is limited, which limits the number of fluorescent agents suitable for the target cells, and it is difficult to detect a mixture of many types of cells. Its resolution decreases. In addition, hematoporphyrin derivatives (HPD), which selectively gather in cancer cells, have recently attracted attention.
), accurate detection becomes impossible because the target cells exhibit bimodal or 3II+\ characteristics. Furthermore, since strong excitation light has a cell-killing effect, there is a limit to the intensity of the excitation light and there are several drawbacks, such as the inability to obtain strong fluorescence.
［発明の目的］
本発明のＩ−１的は、上述のような従来装置の欠点を除
去するために、流れている細胞の蛍光１１１１定Ｉ１１
を間を長くする手段と、連続的に蛍光波長を測定する手
段とを設け、微弱蛍光の分光分布を単−流通部中で繰り
返し測定し、その結果を演算することにより雑音の少な
い精度の高い蛍光の分光分布が得られ、ＡＩ６定対象を
広範囲にできる細胞識別装置を提供することにある。[Objective of the Invention] I-1 of the present invention is to eliminate the drawbacks of the conventional device as described above.
By providing a means for increasing the time interval and a means for continuously measuring the fluorescence wavelength, the spectral distribution of weak fluorescence is repeatedly measured in a single flow section, and the results are calculated, thereby achieving high accuracy with less noise. An object of the present invention is to provide a cell identification device that can obtain a spectral distribution of fluorescence and can target a wide range of AI6 targets.
［発明の４１１要］
この目的を達成するための本発明の要旨は、検体に照射
する励起用光ビームの照射域の形状を、検体の流れの方
向を長辺とし流れに垂直な方向を短辺とする長方形とす
る励起手段と、検体から発する蛍光を前記長辺に沿って
律わ夕的に測光する集光手段と、該集光手段によって得
られる光の分光分布を測定する分光分布測定手段とを具
備することを特徴とする細胞識別装置である。[411 Summary of the Invention] The gist of the present invention to achieve this object is to set the shape of the irradiation area of the excitation light beam irradiated to the specimen so that the long side is in the direction of the flow of the specimen and the short side is in the direction perpendicular to the flow. excitation means having a rectangular shape as sides; light collecting means for uniformly measuring fluorescence emitted from the specimen along the long sides; and spectral distribution measurement for measuring the spectral distribution of light obtained by the light collecting means. A cell identification device characterized by comprising: means.
［発明の実施例］
次に、本発明をｆｔ５ｚ図以下に図示の実施例に基づい
て詳細に説明する。[Embodiments of the Invention] Next, the present invention will be described in detail based on embodiments illustrated below with ft5z diagrams.
本発明の実施例は流れている細胞上にその流れ方向を長
手方向とする長方形の形状に集光するための励起手段、
細胞から発せられる蛍光を測定器に集光するための集光
手段、この集光手段によって集光された蛍光を測定する
ための分光分布測定手段、前方及び側方散乱光を測定す
る散乱光測定子役とから構成されている。An embodiment of the present invention includes an excitation means for focusing light onto flowing cells in a rectangular shape whose longitudinal direction is the flow direction;
A condensing means for concentrating fluorescence emitted from cells onto a measuring device, a spectral distribution measuring means for measuring the fluorescence condensed by the condensing means, and a scattered light measurement for measuring forward and side scattered light. It is made up of child actors.
励起手段は第２図に示すように入射ビームＬａと流通部
Ｓとの間に、Ｚ方向の照射ビームの幅を規定する凹円柱
レンズ１０と凸円柱レンズ１１が光源側から順次に配列
され、更にＸ方向の照射ビ−１、Ｌｂの’Ｎ＋ｆを規ル
°するための凸円柱レンズ１２が設置されている。As shown in FIG. 2, the excitation means includes a concave cylindrical lens 10 and a convex cylindrical lens 11 that define the width of the irradiation beam in the Z direction, which are sequentially arranged from the light source side between the incident beam La and the circulation part S. Furthermore, a convex cylindrical lens 12 is installed for regulating 'N+f' of the irradiation beam 1, Lb in the X direction.
集光手段は流通部Ｓにおいて照射ビームしｂにより励起
された細胞から発せられる蛍光を集光するために、Ｚ方
向に並べたオプティカルファイバ群１３と集光機能を有
する１本のファイバ１４とから成り、ファイ／＜群１３
は末端で１束とされファイバ１４に結合されている。The condensing means includes a group of optical fibers 13 arranged in the Z direction and one fiber 14 having a condensing function in order to condense the fluorescence emitted from the cells excited by the irradiation beam b in the flow section S. becomes, phi/<group 13
are combined into one bundle at the end and coupled to the fiber 14.
分光分布＋！ＩＩＩ定手段は第３図に示すように、分光
Ｗ ｌ ５、チャンネルプレートｌ ６ 、−次元フ第
１・センサアレイ１７とから構成されており、ファイバ
１４は分光器１５に接続され、その射出面にチャンネル
プレート１６が設けられ、更に一次元フオドセンサアレ
イ１７の受光面がチャンネルプレート１６の出力面に接
続されている。Spectral distribution +! As shown in FIG. 3, the III determining means is composed of a spectrometer W l 5, a channel plate l 6, and a -dimensional first sensor array 17, and the fiber 14 is connected to a spectrometer 15, and its output A channel plate 16 is provided on the surface, and a light receiving surface of a one-dimensional food sensor array 17 is connected to the output surface of the channel plate 16.
分光分布測定手段によって得られたデータを演算処理す
るために、センサアレイ１７に高速アナログ・デジタル
変換器１６、記憶加算装置１９゜インターフェイス２０
．ＭＰＵ （マイクロプロセッサユニット）２１、表示
装置２２が順次に直列的に接り′シされている。またイ
ンターフェイス２０には、前方散乱光データ部２３と側
方散乱光データ部２４が接続されている。In order to process the data obtained by the spectral distribution measuring means, the sensor array 17 includes a high-speed analog-to-digital converter 16, a storage adder 19 and an interface 20.
．． An MPU (microprocessor unit) 21 and a display device 22 are sequentially connected in series. Further, a forward scattered light data section 23 and a side scattered light data section 24 are connected to the interface 20.
散乱光測定手段は第４図に示すように、流通部Ｓの近傍
に設置され、照射ビームＬｂの細胞番こより散乱された
散乱光の一部を集光するための対物レンズ２５の直前と
、この対物レンズ２５の焦点位置のそれぞれにリング状
スリット２６．２７が配置され、スリット２７の直後に
受光面の小さなフォトダイオード２８が配置されている
。このＪｕｌｌ定手段は前方散乱光用と側方散乱光用に
２組用し１られ、前方散乱光の測定結果は前方散乱光デ
ータ部２３に、側方散乱光による結果は側方散乱光デー
タ部２４に出力される。As shown in FIG. 4, the scattered light measuring means is installed near the flow section S, and is located immediately before an objective lens 25 for condensing a part of the scattered light scattered by the cell number of the irradiation beam Lb. Ring-shaped slits 26 and 27 are arranged at each focal point of the objective lens 25, and a photodiode 28 with a small light-receiving surface is arranged immediately after the slits 27. Two sets of this Jull determination means are used, one for forward scattered light and one for side scattered light, and the measurement results for forward scattered light are stored in the forward scattered light data section 23, and the results for side scattered light are stored in side scattered light data. It is output to section 24.
励起手段においては、照射ビームＬｂのＺ方向の幅は円
柱レンズ１０、ｌｌにより細胞流の層流が乱れない範囲
で、かつ測定に十分な長さ１例えば１０ｍｍ程度のもの
とされている。Ｘ方向は円柱レンズｔｏ、ｔｔでは入射
ビームＬａと同じ幅で通過し、円柱レンズ１２により従
来例と同程度の幅の約１００〜２００ＪＬｍのビーム幅
とされている。このように、円柱レンズ１０．１１，１
２を組合わせることにより、流通部Ｓに所定の形状の平
面波による照射ビームＬｂが形成される。流通部Ｓを通
過する細胞は、細胞流の圧力、流速、細胞濃度等を調整
することにより、照射ビームＬｂ中をＺ方向に２個以上
重なって入らないようになっている。また、照射ビーム
Ｌｂを通過中の細胞は連続的に励起され、蛍光はほぼ定
常状態で発光されることになる。In the excitation means, the width of the irradiation beam Lb in the Z direction is within a range where the laminar flow of the cell flow is not disturbed by the cylindrical lenses 10 and 11, and the length is sufficient for measurement, for example, about 10 mm. In the X direction, the beam passes through the cylindrical lenses to and tt with the same width as the incident beam La, and the cylindrical lens 12 makes the beam width about 100 to 200 JLm, which is about the same width as the conventional example. In this way, the cylindrical lens 10.11,1
By combining 2, an irradiation beam Lb of a plane wave having a predetermined shape is formed in the circulation portion S. By adjusting the pressure, flow rate, cell concentration, etc. of the cell flow, two or more cells passing through the flow section S are prevented from entering the irradiation beam Lb in an overlapping manner in the Z direction. Further, cells passing through the irradiation beam Lb are continuously excited, and fluorescence is emitted in a substantially steady state.
集光手段においては、オプティカルファイバ群１３の個
々のファイバは、受光域が互いに重なり合うように配列
されている。即ち、励起手段と集光手段とにより、照射
ビームＬｂ中を通過するＭ＋１胞は、常に１１光を発し
、そのイｉ￥光はその位置に応じたオプティカルファイ
バ群↑１３の何れかのファイバで集められ、ファイバ１
４からは連続した”Ｉｔ光がイ１＃られることになる。In the light condensing means, the individual fibers of the optical fiber group 13 are arranged so that their light receiving areas overlap with each other. That is, due to the excitation means and the condensing means, the M+1 cell that passes through the irradiation beam Lb always emits 11 lights, and the 1 light is sent to any one of the optical fiber groups ↑13 depending on its position. collected, fiber 1
From 4 onwards, continuous "It light" will be emitted.
分光分布１１１１ 足手段においては、分光器１５で回
折された波長成分がチャンネルプレー）１６の対応する
チャンネルに入射し、そこで増幅されて一次元フオドセ
ンサアレイ１７の対応する画素に入力される。この一次
元フォトセンサアレイ１７は高速で走査されるため、細
胞が照射ビームＬｂ中を通過中に繰り返し分光データを
得ることができる。一次元フォトセンサアレイ１７の出
力は高速アナログ・デジタル変換器１８によりデジタル
量に変換され、記憶加算装置１９において波長ごとに前
走査データに加算され記憶される。このように、走査ご
との各波長での信号を高速でデジタル変換して記憶して
おき走査ごとに加算すれば、微弱な蛍光であってもその
分光分布特性を高いＳ／Ｎ比で測定することが可能とな
る。Spectral Distribution 1111 In the foot means, the wavelength components diffracted by the spectrometer 15 enter the corresponding channels of the channel plate 16, where they are amplified and input to the corresponding pixels of the one-dimensional food sensor array 17. Since this one-dimensional photosensor array 17 is scanned at high speed, spectral data can be repeatedly obtained while the cells are passing through the irradiation beam Lb. The output of the one-dimensional photosensor array 17 is converted into a digital quantity by a high-speed analog-to-digital converter 18, which is added to the previous scan data for each wavelength in a storage adder 19 and stored. In this way, if the signals at each wavelength for each scan are digitally converted at high speed and stored, and then added for each scan, the spectral distribution characteristics of even weak fluorescence can be measured with a high S/N ratio. becomes possible.
散乱光測定手段においては詳細な説明は省略するが、従
来と同様な０．２〜２度程度の角度の散乱光を他の影響
を受けずに測定し、散乱光データを１１＃ることができ
る。Although a detailed explanation of the scattered light measurement means will be omitted, it is possible to measure scattered light at an angle of about 0.2 to 2 degrees without being influenced by other factors, and to convert the scattered light data into 11#s, similar to conventional methods. can.
照射ビームＬｂ中の細胞の通過が終了した時点で、蛍光
データはインターフェイス２０を介してＭＰＵ２１に取
り込まれる。この終了時点はここでは説明しない検出手
段により、照射ビームしｂ［［１への細胞の出入りを検
知し、その検知信号を用いて容易に知ることができる。When the passage of the cells in the irradiation beam Lb is completed, the fluorescence data is taken into the MPU 21 via the interface 20. This end point can be easily determined by detecting the entry and exit of cells into the irradiation beam b[[1 by a detection means not described here, and using the detection signal.
ＭＰＵ２１では異なるタイミングでインターフェイス２
０を介してｌｉｄ乱光データｔ：１１２３．２４から取
り込んだ散乱光データを、蛍光データと合わせて解析を
行い、その結果を表示装置／（２２に出力する。MPU21 uses interface 2 at different timings.
The scattered light data taken in from the lid scattered light data t: 1123.24 through 0 is analyzed together with the fluorescence data, and the results are output to the display device/(22).
以北のことをまとめると、蛍光標識された細胞が長方形
の照射ビームＬｂ中に入ると、先ず散乱光測定手段によ
り前方散乱光強度、側方散乱光強度が７１１１定され、
インターフェイス２０を介してＭＰＵ２１に取り込まれ
る。一方、照射ビームＬｂ中に細胞が入ったことが検知
されると蛍光分布測定が開始され、一次元フォトセンサ
アレイ１７が繰り返し走査され、走査ごとに得られる蛍
光分光分布を細胞が照射ビームＬｂから抜は出るまで加
算する。その結果、得られた高いＳ／Ｎ比の蛍光分光分
布データが、インターフェイス２０を介してＭＰＵ２１
に取り込まれる。この一連の動作は細胞が照射ビームＬ
ｂ中を通過するごとに繰り返される。ＭＰＵ２１では、
繰り返し得られるｌｉｄ乱光データとイ１？光分光分布
データとを解析し、その結果を表示装置２２に表示する
。To summarize further, when a fluorescently labeled cell enters the rectangular irradiation beam Lb, the forward scattered light intensity and side scattered light intensity are first determined by the scattered light measuring means,
The data is taken into the MPU 21 via the interface 20. On the other hand, when it is detected that a cell has entered the irradiation beam Lb, fluorescence distribution measurement is started, and the one-dimensional photosensor array 17 is repeatedly scanned, and the fluorescence spectral distribution obtained for each scan is detected when the cell is exposed to the irradiation beam Lb. Add the number until it is removed. As a result, the obtained fluorescence spectral distribution data with a high S/N ratio is transmitted to the MPU 21 via the interface 20.
be taken in. This series of operations causes the cells to be exposed to the irradiated beam L.
It is repeated each time it passes through b. In MPU21,
Lid scattering data obtained repeatedly and i1? The optical spectral distribution data is analyzed and the results are displayed on the display device 22.
この装置は上述のような作用を行う励起手段と集光手段
と分光分布Ｉ！１１定手段定石段ていればよく、例えば
集光手段は実施例のように、オプチカルファイバではな
くレンズにより構成することも可能である。This device consists of an excitation means, a light condensing means, and a spectral distribution I! For example, the condensing means may be composed of a lens instead of an optical fiber as in the embodiment.
［発明の効果］
以上説明したように本発明に係る細胞識別装置は、微弱
な蛍光であっても連続的に高いＳ／Ｎ比で分光分布１１
１１定を可能としたことにより、従来の２色法による計
測結果に比較して得られる情報が飛躍的に増大し、測定
対象を広範囲にすることができ、より多種類の細胞が混
在する検体の識別が可能となり、特にＨＰ誘導体による
腫瘍細胞の分類に好適に用いることができる。[Effects of the Invention] As explained above, the cell identification device according to the present invention continuously analyzes the spectral distribution 11 with a high S/N ratio even with weak fluorescence.
By making it possible to measure 11 constants, the amount of information obtained is dramatically increased compared to measurement results using the conventional two-color method, and the measurement target can be expanded to a wider range, allowing samples containing a wider variety of cells to be measured. It can be used particularly for classifying tumor cells based on HP derivatives.
第１図は従来の励起光学系の構成図、第２図以下は本発
明に係る細胞識別装置の一実施例を示し、第２図はその
励起手段及び集光手段の構成図、第３図は分光分布ＡＩ
４定手段のブロック回路構成図、第４図はｌｌ＆乱光測
定手段の構成図である。
符号１０．１１．１２は円柱レンズ、１３はオプティカ
ルファイバ群、１４はファイバ、１５は分光器、１６は
チャンネルプレート、１７は一次元フオドセンサアレイ
、１８は高速アナログ・デジタル変換器、１９は記憶加
算装置、２０はインクフェイス、２１はＭＰＵ、２２は
表示装置、２３は前方散乱光データ部、２４は側方散乱
光データ部、４２５は対物レンズ、２８はフォトダイオ
ードである。
特許出願人 キャノン株式会社
代 理 人 弁理士 Ｕ 比 谷 征 彦゛・１ 。FIG. 1 is a configuration diagram of a conventional excitation optical system, FIG. 2 and the following diagrams show an embodiment of the cell identification device according to the present invention, FIG. is the spectral distribution AI
FIG. 4 is a block circuit diagram of the 4 constant means, and FIG. 10, 11, 12 are cylindrical lenses, 13 is an optical fiber group, 14 is a fiber, 15 is a spectrometer, 16 is a channel plate, 17 is a one-dimensional food sensor array, 18 is a high-speed analog-to-digital converter, and 19 is a 20 is an ink face, 21 is an MPU, 22 is a display device, 23 is a forward scattered light data section, 24 is a side scattered light data section, 425 is an objective lens, and 28 is a photodiode. Patent applicant: Canon Co., Ltd. Representative Patent attorney: Yukihiko Hitani 1.
Claims (1)
検体の流れの方向を長辺とし流れに垂直な方向を短辺と
する長方形とする励起手段と、検体から発する蛍光を前
記長辺に沿って連続的に測光する集光手段と、該集光手
段によって得られる光の分光分布を測定する分光分布測
定手段とを具備することを特徴とする細胞識別装置。 ２、前記照射域の長辺の長さは、検体の層流が乱されな
い範囲とした特許請求の範囲第１項に記載の細胞識別装
置。 ３、前記励起手段は、検体の流れの方向への照射光拡大
光学系と、流れに垂直な方向への照射光集光光学系とか
ら構成した特許請求の範囲第１項に記載の細胞識別装置
。 ４、前記集光手段は、前記長方形照射域の長辺に沿って
直線状に配列したファイバ列と、該ファイバ列の出力を
集束するための他の１本のファイバとから構成した特許
請求の範囲第１項に記載の細胞識別装置。 ５、前記分光分布測定手段は、分光器、光増幅器、一次
元フォトセンサアレイから構成した特許請求の範囲第１
項に記載の細胞識別装置。 ６、前記一次元フォトセンサアレイは高速で繰り返して
走査するようにした特許請求の範囲第５項に記載の細胞
識別装置。[Claims] 1. The shape of the irradiation area of the excitation light beam irradiated onto the specimen is
an excitation means having a rectangular shape with a long side in the direction of the flow of the specimen and a short side in the direction perpendicular to the flow; a light collecting means for continuously measuring fluorescence emitted from the specimen along the long side; and the light collecting means. 1. A cell identification device comprising: spectral distribution measuring means for measuring the spectral distribution of light obtained by the means. 2. The cell identification device according to claim 1, wherein the length of the long side of the irradiation area is within a range where the laminar flow of the sample is not disturbed. 3. Cell identification according to claim 1, wherein the excitation means comprises an optical system for expanding the irradiated light in the direction of the flow of the specimen and an optical system for concentrating the irradiated light in the direction perpendicular to the flow. Device. 4. The light focusing means is constituted by a fiber array arranged in a straight line along the long side of the rectangular irradiation area, and another fiber for focusing the output of the fiber array. The cell identification device according to scope 1. 5. Claim 1, wherein the spectral distribution measuring means comprises a spectrometer, an optical amplifier, and a one-dimensional photosensor array.
The cell identification device described in section. 6. The cell identification device according to claim 5, wherein the one-dimensional photosensor array scans repeatedly at high speed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59174505A JPS6151569A (en) | 1984-08-22 | 1984-08-22 | Cell identifying device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP59174505A JPS6151569A (en) | 1984-08-22 | 1984-08-22 | Cell identifying device |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS6151569A true JPS6151569A (en) | 1986-03-14 |
Family
ID=15979674
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP59174505A Pending JPS6151569A (en) | 1984-08-22 | 1984-08-22 | Cell identifying device |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6151569A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6336147A (en) * | 1986-07-30 | 1988-02-16 | Shimadzu Corp | Gel electrophotographic device |
| JPH03170846A (en) * | 1989-11-09 | 1991-07-24 | General Atomic Co | Micropipette adapter and optical measuring method using the same |
| JPH04335135A (en) * | 1991-05-11 | 1992-11-24 | Horiba Ltd | Particulate counter |
| US5428451A (en) * | 1989-12-07 | 1995-06-27 | Diatec Instruments A/S | Process and apparatus for counting particles |
| US6731100B1 (en) * | 1997-05-05 | 2004-05-04 | Chemometec A/S | Method and a system for determination of somatic cells in milk |
| JP2006250686A (en) * | 2005-03-10 | 2006-09-21 | Mitsui Eng & Shipbuild Co Ltd | Flow site meter and laser beam irradiation method |
| JP2007527997A (en) * | 2004-03-06 | 2007-10-04 | マイケル トレイナー， | Method and apparatus for determining particle size and shape |
| JP2011503551A (en) * | 2007-11-05 | 2011-01-27 | アボット・ラボラトリーズ | Method and apparatus for rapidly counting and identifying biological particles in a stream |
| JP2016522409A (en) * | 2013-06-03 | 2016-07-28 | エックストラリス・テクノロジーズ・リミテッド | Particle detection system and related method |
| JP2019148504A (en) * | 2018-02-27 | 2019-09-05 | シスメックス株式会社 | Particle measuring apparatus and particle measuring method |
-
1984
- 1984-08-22 JP JP59174505A patent/JPS6151569A/en active Pending
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6336147A (en) * | 1986-07-30 | 1988-02-16 | Shimadzu Corp | Gel electrophotographic device |
| JPH03170846A (en) * | 1989-11-09 | 1991-07-24 | General Atomic Co | Micropipette adapter and optical measuring method using the same |
| US5428451A (en) * | 1989-12-07 | 1995-06-27 | Diatec Instruments A/S | Process and apparatus for counting particles |
| JPH04335135A (en) * | 1991-05-11 | 1992-11-24 | Horiba Ltd | Particulate counter |
| US6731100B1 (en) * | 1997-05-05 | 2004-05-04 | Chemometec A/S | Method and a system for determination of somatic cells in milk |
| JP2007527997A (en) * | 2004-03-06 | 2007-10-04 | マイケル トレイナー， | Method and apparatus for determining particle size and shape |
| JP2012103259A (en) * | 2004-03-06 | 2012-05-31 | Michael Trainer | Methods and apparatus for determining size and shape of particles |
| JP2016026301A (en) * | 2004-03-06 | 2016-02-12 | トレイナー， マイケルＴＲＡＩＮＥＲ， Ｍｉｃｈａｅｌ | Method and apparatus for determining size and shape of particles |
| JP2006250686A (en) * | 2005-03-10 | 2006-09-21 | Mitsui Eng & Shipbuild Co Ltd | Flow site meter and laser beam irradiation method |
| JP2011503551A (en) * | 2007-11-05 | 2011-01-27 | アボット・ラボラトリーズ | Method and apparatus for rapidly counting and identifying biological particles in a stream |
| JP2016522409A (en) * | 2013-06-03 | 2016-07-28 | エックストラリス・テクノロジーズ・リミテッド | Particle detection system and related method |
| JP2019148504A (en) * | 2018-02-27 | 2019-09-05 | シスメックス株式会社 | Particle measuring apparatus and particle measuring method |
| WO2019167499A1 (en) * | 2018-02-27 | 2019-09-06 | シスメックス株式会社 | Particle measuring device and particle measuring method |
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