JPH08271509A - Particle sorter - Google Patents
Particle sorterInfo
- Publication number
- JPH08271509A JPH08271509A JP7076890A JP7689095A JPH08271509A JP H08271509 A JPH08271509 A JP H08271509A JP 7076890 A JP7076890 A JP 7076890A JP 7689095 A JP7689095 A JP 7689095A JP H08271509 A JPH08271509 A JP H08271509A
- Authority
- JP
- Japan
- Prior art keywords
- scattered light
- light intensity
- angle
- small
- scattering light
- 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
- 239000002245 particle Substances 0.000 title claims abstract description 38
- 210000000265 leukocyte Anatomy 0.000 claims abstract description 35
- 238000009826 distribution Methods 0.000 claims abstract description 23
- 238000001514 detection method Methods 0.000 claims abstract description 15
- 210000001616 monocyte Anatomy 0.000 claims description 25
- 210000003651 basophil Anatomy 0.000 claims description 22
- 230000001678 irradiating effect Effects 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 abstract description 22
- 239000008187 granular material Substances 0.000 description 15
- 238000010586 diagram Methods 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- 210000004698 lymphocyte Anatomy 0.000 description 7
- 210000000440 neutrophil Anatomy 0.000 description 7
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 206010004173 Basophilia Diseases 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 210000003979 eosinophil Anatomy 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 1
- 208000032791 BCR-ABL1 positive chronic myelogenous leukemia Diseases 0.000 description 1
- 208000010833 Chronic myeloid leukaemia Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 206010020850 Hyperthyroidism Diseases 0.000 description 1
- 201000005505 Measles Diseases 0.000 description 1
- 201000003793 Myelodysplastic syndrome Diseases 0.000 description 1
- 208000033761 Myelogenous Chronic BCR-ABL Positive Leukemia Diseases 0.000 description 1
- 231100000354 acute hepatitis Toxicity 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000010419 fine particle Substances 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、例えば白血球細胞等の
粒子に光を照射して分類する粒子分類装置に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a particle classification device for irradiating particles such as white blood cells with light for classification.
【０００２】[0002]
【従来の技術】一般に、白血球細胞は、リンパ球、単
球、好中球、好酸球、好塩基球等の種々の細胞から成
る。これら各細胞は核と細胞質から成り、各細胞は、大
きさ、形状が異なると共に、細胞質に含まれる顆粒等の
微粒子の大きさ・形状・量等が夫々異なっていることが
知られている。上記各細胞の含有率及び各細胞中に含ま
れる顆粒又は粒子に関しては以下に示すことが判明して
いる。 リンパ球（２５〜４５％）；顆粒がほとんどない。 単球（４〜７％） ；０．１μｍ以下の顆粒が均
一に極めて多数存在する。 好中球（４５〜６１％） ；０．０５〜０．２μｍのグ
リコーゲン粒子が細胞全体に多数存在する。 好酸球（１〜５％） ；０．５〜１．０μｍの特殊
顆粒が多数存在する。 好塩基球（０〜１％） ；０．２〜１．０μｍの顆粒
が多数存在する。 （ ）内の数値は、各細胞の含有率を示す。 これらの各細胞が疾患により増減するので、これらの細
胞中の顆粒又は粒子から得られる情報に基づき、各細胞
の状態を検出することにより疾患の判定や診断が行わ
れ、白血球の分類は臨床検査において有益なものであ
る。2. Description of the Related Art Generally, white blood cells consist of various cells such as lymphocytes, monocytes, neutrophils, eosinophils and basophils. It is known that each of these cells is composed of a nucleus and a cytoplasm, and that each cell has a different size and shape, and that the size, shape, amount, etc. of fine particles such as granules contained in the cytoplasm are different. The content of each cell and the granules or particles contained in each cell are known to be as follows. Lymphocytes (25-45%); few granules. Monocytes (4-7%); Granules of 0.1 μm or less are uniformly present in a very large number. Neutrophils (45-61%); glycogen particles of 0.05-0.2 μm are present in large numbers throughout the cells. Eosinophils (1-5%); There are many special granules of 0.5-1.0 μm. Basophils (0-1%); There are many granules of 0.2-1.0 μm. The numerical value in () shows the content rate of each cell. Since each of these cells increases or decreases depending on the disease, the disease is judged or diagnosed by detecting the state of each cell based on the information obtained from the granules or particles in these cells. Is useful in.
【０００３】従来、これら白血球細胞にレーザ光を照射
し、細胞の大きさを反映する前方散乱光及び細胞の顆粒
特性等を表す側方散乱光の２方向の光を検出して分析す
る細胞分析装置が一般に多用されている。Conventionally, cell analysis in which these white blood cells are irradiated with laser light to detect and analyze forward scattered light that reflects the size of the cell and side scattered light that represents the granular characteristics of the cell, etc. Devices are commonly used.
【０００４】各細胞内には、使用されるレーザ光の波長
に比較して、上述したように大きな顆粒又は粒子から小
さな顆粒又は粒子が含まれているため、レーザ光照射時
に散乱する散乱光には以下に示すような方向性がある。 顆粒又は粒子の大きさが波長に対して大きい場合 ；前
方散乱光 顆粒又は粒子の大きさが波長と同程度の場合 ；前
方及び側方散乱光 顆粒又は粒子の大きさが波長に対して小さい場合 ；前
方、側方及び後方散乱光As described above, each cell contains large granules or small particles or small granules or particles as compared with the wavelength of the laser light used. Has the following directions. When the size of the granules or particles is large with respect to the wavelength; Forward scattered light When the size of the granules or particles is similar to the wavelength; When the size of the forward and side scattered light granules or particles is small with respect to the wavelength ; Forward, side and back scattered light
【０００５】白血球細胞に含まれる顆粒又は粒子の大き
さが異なることにより、前述した従来の前方及び側方散
乱光のみによる分析では、必ずしも各細胞が明確に分離
識別できないため、本願出願人は、前方及び側方散乱光
に加えて後方散乱光による粒子分類装置を提案した（特
願平６−２６８９１７）。Since the size of the granules or particles contained in the white blood cells is different, it is not always possible to clearly separate and identify each cell by the above-mentioned conventional analysis using only forward and side scattered light. We proposed a particle classification device using backscattered light in addition to forward and side scattered light (Japanese Patent Application No. 6-268917).
【０００６】図５は、本願出願人が提案した粒子分類装
置の概略図である。図５において、レーザ光源１からレ
ーザ光が照射光集束用のレンズ２、３、中央部に孔を設
けた後方散乱光を反射するミラー１３及び同様の孔を有
するコリメート用レンズ１２を介して流動室４に照射さ
れる。レーザ光は、流動室４でサンプル液に含まれる白
血球細胞により反射、透過或いは回析などにより散乱さ
れ、散乱光が前方散乱光集束用レンズ８を介して検出器
９に入射され、電気信号に変換されて出力される。この
前方散乱光集束用レンズ８及び検出器９により前方散乱
光検出手段を構成する。FIG. 5 is a schematic diagram of a particle classification apparatus proposed by the applicant of the present application. In FIG. 5, the laser light from the laser light source 1 flows through the lenses 2 and 3 for focusing the irradiation light, the mirror 13 for reflecting the backscattered light having a hole in the center, and the collimating lens 12 having the similar hole. The chamber 4 is irradiated. The laser light is scattered by the white blood cells contained in the sample liquid in the flow chamber 4 by reflection, transmission or diffraction, and the scattered light is incident on the detector 9 through the forward scattered light focusing lens 8 and converted into an electric signal. It is converted and output. The forward scattered light focusing lens 8 and the detector 9 constitute forward scattered light detecting means.
【０００７】上記流動室４は、概略的に図６Ａに示すよ
うに構成されている。内腔が漏斗状の容器４ａ内に、そ
の外周部に沿ってシース液（鞘液）Ｓｗが流れ、中央部
を分析しようとする白血球細胞を含むサンプル液Ｆが流
れる。この場合、シース液Ｓｗ及びサンプル液Ｆは、レ
イノルズの原理によりその流速の違いから混合しないよ
うになっている。サンプル液Ｆは、細管部４ｂを通過す
る際、レーザ光が照射される。The flow chamber 4 is generally constructed as shown in FIG. 6A. A sheath liquid (sheath liquid) Sw flows along the outer periphery of the container 4a having a funnel-shaped lumen, and a sample liquid F containing white blood cells to be analyzed flows in the central part. In this case, the sheath liquid Sw and the sample liquid F are not mixed due to the difference in their flow velocities due to the Reynolds principle. The sample liquid F is irradiated with laser light when passing through the thin tube portion 4b.
【０００８】また、図６Ｂは、細管部４ｂをレーザ光が
サンプル液Ｆ内の白血球細胞Ｃに入射する詳細を示し、
ここでレーザ光は白血球細胞Ｃにより散乱されて、上述
した前方散乱光、側方散乱光及び後方散乱光が、夫々の
検出手段により検出される。FIG. 6B shows the details of the laser light incident on the white blood cells C in the sample liquid F through the thin tube portion 4b.
Here, the laser light is scattered by the white blood cells C, and the above-mentioned forward scattered light, side scattered light, and back scattered light are detected by the respective detection means.
【０００９】また、流動室４で直角方向に散乱された散
乱光をを、側方散乱光集束用レンズ５及び検出器６で構
成される側方散乱光検出手段により検出され、電気信号
に変換されて出力される。The scattered light scattered in the flow chamber 4 at right angles is detected by the side scattered light detecting means composed of the side scattered light focusing lens 5 and the detector 6, and converted into an electric signal. Is output.
【００１０】更に、流動室４でレーザ光の入射方向と逆
方向に散乱される後方散乱光は、ミラー１３、後方散乱
光集束用レンズ１４及び検出器１５で構成される後方散
乱光検出手段により検出され、電気信号に変換されて出
力される。Further, the backscattered light scattered in the flow chamber 4 in the direction opposite to the incident direction of the laser light is detected by the backscattered light detecting means composed of the mirror 13, the backscattered light focusing lens 14 and the detector 15. It is detected, converted into an electric signal, and output.
【００１１】各検出器９、６及び１５により検出された
前方、側方及び後方散乱光に対応する検出信号は分析装
置１０に入力されて記憶され、夫々の散乱光強度が算定
されて表示装置１１に出力される。表示装置１１は、前
方、側方及び後方散乱強度データを画面上の２次元座標
上に表示する。そして、前方、側方、後方の散乱光強度
を組み合わせて２次元座標上にスキャッタグラムとして
表示することにより、白血球に含まれる各細胞の分布か
ら分類を行うことができる。The detection signals corresponding to the forward, side and back scattered light detected by each of the detectors 9, 6 and 15 are input to and stored in the analyzer 10, the respective scattered light intensities are calculated, and the display device is displayed. 11 is output. The display device 11 displays the front, side, and backscattering intensity data on the two-dimensional coordinates on the screen. Then, the scattered light intensities of the front, side, and rear are combined and displayed as a scattergram on the two-dimensional coordinates, so that classification can be performed from the distribution of each cell contained in the white blood cells.
【００１２】このように、上述した従来の粒子分類装置
では、白血球細胞から散乱した前方散乱光、側方散乱光
及び後方散乱光を検出して小さな顆粒又は粒子を含む白
血球細胞の分類を行うようにしていた。As described above, in the above-described conventional particle classification device, the forward scattered light, the side scattered light, and the back scattered light scattered from the white blood cells are detected to classify the white blood cells containing small granules or particles. I was doing.
【００１３】[0013]
【発明が解決しようとする課題】しかしながら、小さな
顆粒又は粒子を含む白血球細胞を前方、側方及び後方散
乱光を検出することによりある程度の細胞の分類を行う
ことができるが、単球や好塩基球などの含有率の低い細
胞は含有率の高いリンパ球や好中球に埋もれて明確に識
別或いは分類することが困難であった。従って、本発明
は上記課題に鑑みてなされたもので、比較的含有率の少
ない細胞を精度良く分類できる粒子分類装置を提供する
ことを目的とする。However, white blood cells containing small granules or particles can be classified to some extent by detecting forward, side, and backscattered light. However, monocytes and basophils can be classified. Cells with a low content such as spheres were buried in lymphocytes and neutrophils with a high content, and it was difficult to clearly identify or classify them. Therefore, the present invention has been made in view of the above problems, and an object of the present invention is to provide a particle classification device capable of accurately classifying cells having a relatively low content rate.
【００１４】[0014]
【課題を解決するための手段】本発明の請求項１に係る
粒子分類装置は、粒子に光を照射し、この粒子による散
乱光を検出して該粒子の分類を行う粒子分類装置におい
て、前方小角散乱光を検出する前方散乱光検出手段と、
前方散乱光検出手段と側方散乱光検出手段との間に配置
され、前方大角散乱光を検出する前方大角散乱光検出手
段と、前方小角、前方大角及び側方散乱光検出手段から
出力される、前方小角散乱光、側方散乱光及び前方大角
散乱光に対応する検出信号を入力して夫々の散乱光強度
データを算定して記憶する記憶手段と、前方小角、前方
大角及び側方散乱光強度データから前方小角散乱光強度
と前方大角散乱光強度の２次元分布データ並びに前方小
角散乱光強度と側方散乱光強度の２次元分布データを算
定する手段と、前方小角散乱光強度と前方大角散乱光強
度の２次元分布データから特定の白血球細胞の領域を決
定する手段とを具える分析手段と、算定された前方小角
散乱光強度と前方大角散乱光強度の２次元分布データ、
前方小角散乱光強度と側方散乱光強度の２次元分布デー
タ並びに特定された白血球細胞のデータを前方小角散乱
光強度と側方散乱光強度の２次元座標上に表示する表示
手段とにより構成される。A particle classifying apparatus according to claim 1 of the present invention is a particle classifying apparatus for illuminating particles and detecting scattered light by the particles to classify the particles. Forward scattered light detection means for detecting small angle scattered light,
It is arranged between the forward scattered light detecting means and the side scattered light detecting means, and is output from the front large angle scattered light detecting means for detecting the front large angle scattered light and the front small angle, front large angle and side scattered light detecting means. Storage means for inputting the detection signals corresponding to the front small-angle scattered light, the side scattered light, and the front large-angle scattered light to calculate and store the scattered light intensity data, and the front small-angle, front large-angle, and side scattered light. Means for calculating two-dimensional distribution data of forward small-angle scattered light intensity and forward large-angle scattered light intensity and two-dimensional distribution data of forward small-angle scattered light intensity and side-scattered light intensity from intensity data, and forward small-angle scattered light intensity and forward large-angle scattered light intensity Analysis means having means for determining a specific white blood cell region from the two-dimensional distribution data of scattered light intensity, and two-dimensional distribution data of the calculated forward small angle scattered light intensity and forward large angle scattered light intensity,
The display means for displaying the two-dimensional distribution data of the front small-angle scattered light intensity and the side scattered light intensity and the data of the identified white blood cells on the two-dimensional coordinates of the front small-angle scattered light intensity and the side scattered light intensity. It
【００１５】請求項２に係る発明は、請求項１記載の粒
子分類装置において、特定の白血球細胞を含有率の少な
い白血球細胞としたことを特徴とする。The invention according to claim 2 is the particle classification apparatus according to claim 1, characterized in that specific white blood cells are white blood cells having a low content rate.
【００１６】請求項３に係る発明は、請求項１又は２記
載の粒子分類装置において、含有率の少ない白血球細胞
を単球及び／又は好塩基球としたことを特徴とする。The invention according to claim 3 is the particle classification apparatus according to claim 1 or 2, wherein the white blood cells having a low content rate are monocytes and / or basophils.
【００１７】[0017]
【作用】請求項１に係る発明では、白血球細胞にレーザ
光を照射して分類を行う際、前方散乱光検出手段により
前方小角散乱光を検出し、前方大角散乱光検出手段によ
り前方大角散乱光を検出すると共に側方散乱光検出手段
により側方散乱光を検出する。分析手段により、側方、
前方及び前方大角散乱光検出手段から出力される、前方
小角、側方及び前方大角の夫々の散乱光強度データを記
憶し、前方小角散乱光強度と前方大角散乱光強度の２次
元分布データ並びに前方小角散乱光強度と側方散乱光強
度の２次元分布データを算定し、前方小角散乱光強度と
前方大角散乱光強度の２次元分布データから特定の白血
球細胞の領域を決定して前方小角及び側方散乱光強度の
２次元分布データとして出力する。算定された前方小角
散乱光強度と前方大角散乱光強度の２次元分布データ、
前方小角散乱光強度と側方散乱光強度の２次元分布デー
タ並びに特定された白血球細胞の散乱光強度データを、
表示手段の画面上に２次元座標上に表示する。In the invention according to claim 1, when the white blood cells are irradiated with laser light for classification, the forward scattered light detecting means detects the forward small angle scattered light, and the forward large angle scattered light detecting means detects the forward large angle scattered light. And the side scattered light is detected by the side scattered light detecting means. Sideways, by analysis means,
The scattered light intensity data for each of the small front angle, the lateral wide angle, and the large front scattered angle output from the front and front large angle scattered light detection means are stored, and the two-dimensional distribution data of the small front scattered angle light intensity and the large forward scattered angle light and the forward direction are stored. Two-dimensional distribution data of small-angle scattered light intensity and side-scattered light intensity are calculated, and a specific white blood cell region is determined from the two-dimensional distribution data of front small-angle scattered light intensity and front large-angle scattered light intensity to determine the front small-angle and side It is output as two-dimensional distribution data of the scattered light intensity. Two-dimensional distribution data of the calculated forward small-angle scattered light intensity and forward large-angle scattered light intensity,
The two-dimensional distribution data of the front small angle scattered light intensity and the side scattered light intensity and the scattered light intensity data of the identified white blood cells,
The two-dimensional coordinates are displayed on the screen of the display means.
【００１８】請求項２に係る発明では、含有率の少ない
白血球細胞を特定の白血球細胞として選択する。In the invention according to claim 2, white blood cells having a low content rate are selected as specific white blood cells.
【００１９】請求項３に係る発明では、含有率の少ない
白血球細胞として単球又は好塩基球或いは両者を選択す
る。In the invention according to claim 3, monocytes or basophils or both are selected as the white blood cells having a low content.
【００２０】[0020]
【実施例】図面を参照して、本発明の粒子分類装置の実
施例について説明する。図１は、本発明の実施例の構成
を示すブロック図である。図１において、図５と対応す
る部分には同一の符号又は類似の符号を付した。DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS An embodiment of the particle classification apparatus of the present invention will be described with reference to the drawings. FIG. 1 is a block diagram showing the configuration of an embodiment of the present invention. In FIG. 1, portions corresponding to those in FIG. 5 are designated by the same reference numerals or similar reference numerals.
【００２１】本例は、図５に示した後方散乱光検出用レ
ンズ５及び検出器６から成る側方散乱光検出手段と、中
央部に孔を有し、後方散乱光を反射するミラー１３、後
方散乱光検出用レンズ１４及び検出器１５から成る後方
散乱光検出手段との位置を入れ替え、後方散乱光検出手
段を前方大角散乱光検出手段として用い、かつ前方散乱
光検出手段を前方小角検出手段として用いるように構成
したものである。その他の構成は、図５に示した従来例
と同様であり、重複説明は省略する。In this example, the side scattered light detecting means consisting of the back scattered light detecting lens 5 and the detector 6 shown in FIG. 5 and the mirror 13 having a hole in the center and reflecting the back scattered light, The position of the backscattered light detecting means composed of the backscattered light detecting lens 14 and the detector 15 is exchanged, the backscattered light detecting means is used as the front large angle scattered light detecting means, and the front scattered light detecting means is the front small angle detecting means. It is configured to be used as. Other configurations are the same as those of the conventional example shown in FIG. 5, and thus redundant description will be omitted.
【００２２】即ち、中央に孔を有し、前方大角散乱光を
反射するミラー１３ａ、このミラー１３ａにより反射さ
れた前方大角散乱光を集束する前方大角散乱光集束用レ
ンズ１４ａ及び前方大角散乱光を電気信号に変換する検
出器１５ａにより前方大角散乱光検出手段が構成され
る。That is, a mirror 13a having a hole in the center and reflecting the front large-angle scattered light, a front large-angle scattered light focusing lens 14a for converging the front large-angle scattered light reflected by the mirror 13a, and a front large-angle scattered light are provided. The detector 15a for converting into an electric signal constitutes a forward large angle scattered light detecting means.
【００２３】次に、前述した本発明の粒子分類装置の動
作について説明する。レーザ光源１から照射されるレー
ザ光は、レーザ光集束用レンズ２及び３に集束され、流
動室４の細管部４ｂ（図６）を通過する白血球細胞Ｃ
（図６）に当たり、側方に散乱した散乱光成分が側方散
乱光検出用レンズ５により検出されて検出器６に入力さ
れる。検出器６から、側方散乱光成分が電気信号に変換
されて側方散乱光強度に対応する検出信号として出力さ
れる。Next, the operation of the above-described particle classification device of the present invention will be described. The laser light emitted from the laser light source 1 is focused on the laser light focusing lenses 2 and 3, and the white blood cell C passing through the thin tube portion 4b (FIG. 6) of the flow chamber 4
In (FIG. 6), the scattered light component scattered sideways is detected by the side scattered light detection lens 5 and input to the detector 6. From the detector 6, the side scattered light component is converted into an electric signal and output as a detection signal corresponding to the side scattered light intensity.
【００２４】また、流動室４の前方（図の左方）に入射
される直射光は照射光ストッパ７により遮蔽され、直射
光以外の散乱光が散乱光用コリメートレンズ１２により
平行光線とされ、ミラー１３ａにより反射される。反射
された散乱光は、前方大角散乱光成分として前方大角散
乱光検出用レンズ１４ａで検出されて検出器１５ａに入
力され、電気信号に変換される。検出器１５ａからは、
前方大角散乱光強度に対応する検出信号として出力され
る。Direct light incident on the front side of the flow chamber 4 (on the left side in the figure) is blocked by the irradiation light stopper 7, and scattered light other than the direct light is made into parallel rays by the scattered light collimating lens 12. It is reflected by the mirror 13a. The reflected scattered light is detected by the front large-angle scattered light detection lens 14a as a front large-angle scattered light component, is input to the detector 15a, and is converted into an electric signal. From the detector 15a,
It is output as a detection signal corresponding to the front large-angle scattered light intensity.
【００２５】更に、ミラー１３の中央部の孔を通過した
前方小角散乱光成分は、前方散乱光検出用レンズ８によ
り検出されて検出器９に入力される。検出器９は、前方
小角散乱光成分を電気信号に変換して前方小角散乱光強
度に対応する検出信号として出力する。Further, the front small-angle scattered light component that has passed through the hole in the center of the mirror 13 is detected by the front scattered light detecting lens 8 and input to the detector 9. The detector 9 converts the front small-angle scattered light component into an electric signal and outputs it as a detection signal corresponding to the front small-angle scattered light intensity.
【００２６】上述した各検出器６、９、１５ａは、夫々
例えばフォトダイオード等の光電変換素子により構成さ
れる。Each of the detectors 6, 9 and 15a described above is composed of a photoelectric conversion element such as a photodiode.
【００２７】検出器６、９、１５ａにより検出された前
方小角、側方及び前方大角散乱光に応じた検出信号は分
析装置１０に入力される。分析装置１０は、入力された
各検出信号から前方小角散乱光強度データ、側方散乱光
強度データ及び前方大角散乱光強度データを算定して記
憶し、且つ前方小角及び前方大角散乱光強度の２次元座
標データ、前方小角散乱光強度と側方散乱光強度の２次
元座標データを算定する。また、分析装置１０は、算定
された前方小角散乱光強度データと前方大角散乱光強度
データ、及び前方小角散乱光強度データと側方散乱光強
度データを順次出力し、或いは分析装置１０の操作部
（図示せず）の操作により表示装置１１に出力し、画面
上に２次元座標の光強度散乱分布図（スキャッタグラ
ム）として表示させる。Detection signals corresponding to the front small angle, side and front large angle scattered lights detected by the detectors 6, 9, 15a are input to the analyzer 10. The analyzer 10 calculates and stores the front small-angle scattered light intensity data, the side scattered light intensity data, and the front large-angle scattered light intensity data from the input detection signals, and stores the front small-angle and front large-angle scattered light intensities. Dimensional coordinate data, two-dimensional coordinate data of small-angle forward scattered light intensity and side scattered light intensity are calculated. Further, the analysis device 10 sequentially outputs the calculated front small-angle scattered light intensity data, front large-angle scattered light intensity data, and front small-angle scattered light intensity data and side scattered light intensity data, or the operation unit of the analysis device 10. It is output to the display device 11 by an operation (not shown) and is displayed on the screen as a two-dimensional coordinate light intensity scattering distribution diagram (scattergram).
【００２８】図２は、上記実施例によるスキャッタグラ
ムを示す。図２Ａ、前方小角散乱光強度を縦軸にとり前
方大角散乱光強度を横軸にとって、例えばリンパ球、単
球、好中球、好塩基球を含むスキャッタグラムである。
また、図２Ｂは、前方小角散乱光強度（縦軸）及び側方
散乱光強度（横軸）による、図２Ａの各白血球細胞のス
キャッタグラムである。FIG. 2 shows a scattergram according to the above embodiment. FIG. 2A is a scattergram including, for example, lymphocytes, monocytes, neutrophils, and basophils with the front small angle scattered light intensity on the vertical axis and the front large angle scattered light intensity on the horizontal axis.
Further, FIG. 2B is a scattergram of each white blood cell in FIG. 2A by the front small angle scattered light intensity (vertical axis) and the side scattered light intensity (horizontal axis).
【００２９】図２Ａ及び図２Ｂのスキャッタグラムで
は、リンパ球、単球、好中球、好塩基球などの各細胞の
分布が混在して表示され、各細胞間のある程度の識別は
可能とであるが、含有率の少ない例えば単球及び好塩基
球の識別は十分ではない。In the scattergrams of FIGS. 2A and 2B, the distributions of cells such as lymphocytes, monocytes, neutrophils, and basophils are displayed in a mixed manner, and it is possible to distinguish each cell to some extent. However, the discrimination of monocytes and basophils having a low content rate is not sufficient.
【００３０】一般に、単球は、免疫機能があり、これが
増加している場合は、結核、麻疹、急性肝炎等の疾患が
あると判断され、また、好塩基球は、増加した場合、甲
状腺機能低下や慢性骨髄性白血病などの疑いがあり、低
下すると甲状腺機能高進症などの疾患があると見做され
る。従って、前述したように、たとえ含有率が小さくて
も単球或いは好塩基球又は両者を分離識別することは重
要である。In general, monocytes have an immune function, and if they are increased, it is judged to have diseases such as tuberculosis, measles and acute hepatitis, and if basophils are increased, thyroid function is increased. There is a suspicion that there is a decline or chronic myelogenous leukemia, and if it falls, it is considered that there is a disease such as hyperthyroidism. Therefore, as described above, it is important to separate and identify monocytes or basophils or both, even if the content is small.
【００３１】図２Ａ及び図２Ｂに示した通常検体のスキ
ャッタグラムでは、例えば単球と好塩基球は混在してい
るため、両者を明確に識別することはできないので、前
方小角散乱光強度及び前方大角散乱光強度のスキャッタ
グラムを表示して、図２Ｃに示すように、単球と好塩基
球が混在した領域（実線で囲んだ領域）を決定し、分析
装置１０により、単球及び好塩基球の散乱光強度データ
のみを表示装置１１に出力する。In the scattergrams of the normal sample shown in FIGS. 2A and 2B, for example, monocytes and basophils are mixed, so that they cannot be clearly discriminated from each other. The scattergram of the large-angle scattered light intensity is displayed, and as shown in FIG. 2C, a region where monocytes and basophils are mixed (a region surrounded by a solid line) is determined, and the analyzer 10 analyzes the monocytes and basophils. Only the scattered light intensity data of the sphere is output to the display device 11.
【００３２】この場合、前方散乱光強度（縦軸）と側方
散乱光強度（横軸）のスキャッタグラムを表示すること
により、図２Ｄに示すように、単球と好塩基球の散乱光
強度データが明確に分離識別することができる。図２Ｃ
に示した領域を示す線と図２Ｄに示した単球領域と好塩
基球領域を識別する線は正常白血球のスキャッタグラム
によって決まるもので、予め記憶された制御プログラム
により自動的に指定することができる。In this case, by displaying the scattergram of the forward scattered light intensity (vertical axis) and the side scattered light intensity (horizontal axis), as shown in FIG. 2D, the scattered light intensity of monocytes and basophils is shown. The data can be clearly separated and identified. Figure 2C
The line indicating the region shown in Fig. 2 and the line identifying the monocyte region and the basophil region shown in Fig. 2D are determined by the scattergram of normal leukocytes, and can be automatically designated by a control program stored in advance. it can.
【００３３】また、図３は、例えば、ＭＤＳ患者（骨髄
異形成症候群）の好塩基球増多検体のスキャッタグラム
で、図２と同様にいずれも前方散乱光強度を縦軸にと
り、前方大角散乱光強度及び側方散乱光強度を横軸とし
ている．図３Ａは、リンパ球、単球、好中球、好塩基球
が混在した前方小角散乱光強度と前方大角散乱光強度の
スキャッタグラムであり、図３Ｂは、図３Ａの前方小角
散乱光強度と側方散乱光強度によるスキャッタグラムで
ある。Further, FIG. 3 is, for example, a scattergram of a basophilia specimen of an MDS patient (myelodysplastic syndrome), and in the same manner as FIG. 2, the forward scattered light intensity is plotted on the vertical axis and the forward large angle scattering is shown. The horizontal axis is the light intensity and side scattered light intensity. FIG. 3A is a scattergram of forward small-angle scattered light intensity and forward large-angle scattered light intensity in which lymphocytes, monocytes, neutrophils, and basophils are mixed, and FIG. 3B is a forward small-angle scattered light intensity of FIG. 3A. It is a scattergram by side scattered light intensity.
【００３４】図３Ｃは、図３Ａの前方小角散乱光強度と
前方大角散乱光強度のスキャッタグラムから、単球及び
好塩基球の混在領域を指定する線を示すものである。ま
た、図３Ｄは、図３Ｃにより指定された領域から、前方
小角散乱光強度と側方散乱光強度とのスキャッタグラム
をとり、好塩基球の領域を識別した好塩基球増多検体の
スキャッタグラムである。FIG. 3C shows a line designating a mixed region of monocytes and basophils from the scattergram of the small forward angle scattered light intensity and the large forward angle scattered light intensity of FIG. 3A. Further, FIG. 3D is a scattergram of the basophil-enhancing specimen in which the forward small-angle scattered light intensity and the side scattered light intensity are taken from the region designated by FIG. 3C and the basophil region is identified. Is.
【００３５】図４は、例えば単球分離検体のスキャッタ
グラムを示すものであり、単球分離検体の分離方法とし
て次に示す方法がある。抗凝固処理した血液と６％（W/
V)Ｄｅｘｔｒａｎ500 を含む生理食塩水を10：１に混合
し、温室で４０分間静置後、血漿層を採取する。これ
を、ＮｙｃｏＰｒｅｐ（登録商標名）1.068 の上に重層
し、 600Ｇで15分間遠心分離して、ＮｙｃｏＰｒｅｐ
（登録商標名）の上層から単球浮遊液を採取する。単球
浮遊液を0.13％（W/V)ＥＤＴＡと１％ＦＣＳを含む生理
食塩水で希釈し、 600Ｇで７分間遠心分離して、洗浄を
する。調整したサンプルは、 0.2μｍのフィルタを通し
て血小板を除去した自己血漿に浮遊させて、単球分離検
体とする。FIG. 4 shows, for example, a scattergram of a monocyte-separated sample, and the following method is available as a method for separating the monocyte-separated sample. Anticoagulated blood and 6% (W /
V) Physiological saline containing Dextran500 is mixed at 10: 1, and after standing still in a greenhouse for 40 minutes, a plasma layer is collected. This was overlaid onto NycoPrep® 1.068 and centrifuged at 600 G for 15 minutes to give NycoPrep
Collect the monocyte suspension from the upper layer (registered trademark). The monocyte suspension is diluted with physiological saline containing 0.13% (W / V) EDTA and 1% FCS, centrifuged at 600 G for 7 minutes, and washed. The prepared sample is suspended in autologous plasma from which platelets have been removed through a 0.2 μm filter to obtain a monocyte separation sample.
【００３６】図４Ａは、リンパ球、単球、好中球、好塩
基球が混在した前方小角散乱光強度（縦軸）と前方大角
散乱光強度（横軸）のスキャッタグラムであり、図４Ｂ
は、図４Ａの前方小角散乱光強度（縦軸）と側方散乱光
強度（横軸）によるスキャッタグラムである。FIG. 4A is a scattergram of forward small-angle scattered light intensity (vertical axis) and forward large-angle scattered light intensity (horizontal axis) in which lymphocytes, monocytes, neutrophils, and basophils are mixed, and FIG.
4A is a scattergram of the front small-angle scattered light intensity (vertical axis) and the side scattered light intensity (horizontal axis) in FIG. 4A.
【００３７】また、図４Ｃは、図４Ａの前方小角散乱光
強度と前方大角散乱光強度のスキャッタグラムから、単
球及び好塩基球の混在領域を指定する線を示ものであ
る。また、図４Ｄは、図４Ｃにより指定された領域か
ら、前方小角散乱光強度と側方散乱光強度とのスキャッ
タグラムをとり、単球の領域を識別したスキャッタグラ
ムである。Further, FIG. 4C shows a line designating a mixed region of monocytes and basophils from the scattergram of the small forward angle scattered light intensity and the large forward angle scattered light intensity of FIG. 4A. Further, FIG. 4D is a scattergram in which a monocyte region is identified by taking a scattergram of the forward small-angle scattered light intensity and the side scattered light intensity from the region designated by FIG. 4C.
【００３８】[0038]
【発明の効果】以上説明したように、請求項１記載の本
発明によれば、前方小角散乱光強度、前方大角散乱光強
度、側方散乱光強度のスキャッタグラムを用いることに
より、白血球細胞内の種々の細胞を明確に分離識別する
ことができる。As described above, according to the present invention as set forth in claim 1, by using the scattergram of the forward small angle scattered light intensity, the forward large angle scattered light intensity and the side scattered light intensity, It is possible to clearly separate and discriminate various cells of.
【００３９】また、請求項２記載の本発明によれば、含
有率の少ない白血球細胞の分離識別が容易となる。Further, according to the present invention as defined in claim 2, white blood cells having a low content rate can be easily separated and identified.
【００４０】更に、請求項３記載の本発明によれば、単
球又は好塩基球或いは両者の分離識別が明確になる利点
がある。Furthermore, according to the present invention as defined in claim 3, there is an advantage that the distinction between monocytes, basophils or both can be clearly identified.
【図１】本発明の粒子分類装置の実施例の構成を示すブ
ロック図である。FIG. 1 is a block diagram showing a configuration of an embodiment of a particle classification device of the present invention.
【図２】図１の実施例による通常検体のスキャッタグラ
ムを示す図である。FIG. 2 is a diagram showing a scattergram of a normal sample according to the embodiment of FIG.
【図３】図１の実施例によるＭＤＳ患者の好塩基球増多
検体のスキャッタグラムを示す図である。FIG. 3 is a diagram showing a scattergram of a basophilia specimen of an MDS patient according to the embodiment of FIG.
【図４】図１の実施例による単球分離検体のスキャッタ
グラムを示す図である。4 is a diagram showing a scattergram of a monocyte-separated specimen according to the embodiment of FIG. 1. FIG.
【図５】従来例の構成を示すブロック図である。FIG. 5 is a block diagram showing a configuration of a conventional example.
【図６】本発明及び従来例の粒子分類装置に使用される
流動室の細部を示す図である。FIG. 6 is a view showing details of a flow chamber used in the particle sorting apparatuses of the present invention and the conventional example.
１ レーザ光源 ２、３ 照射光収束用のレンズ ４ 流動室 ５ 側方散乱光検出用レンズ ８ 前方散乱光検出用レンズ ６、９、１５ａ 検出器 ７ 照射光ストッパ １０ 分析装置（分析手段） １１ 表示装置（表示手段） １２ 散乱光用コリメートレンズ １３ａ ミラー １４ａ 前方大角散乱光検出用レンズ DESCRIPTION OF SYMBOLS 1 Laser light source 2, 3 Lens for converging irradiation light 4 Flow chamber 5 Lens for detecting side scattered light 8 Lens for detecting forward scattered light 6, 9, 15a Detector 7 Irradiation light stopper 10 Analyzing device (analyzing means) 11 Display Device (display means) 12 Collimated lens for scattered light 13a Mirror 14a Lens for detecting large front angle scattered light
Claims (3)
を検出して該粒子の分類を行う粒子分類装置において、 前方小角散乱光を検出する前方散乱光検出手段と、 上記前方散乱光検出手段と側方散乱光検出手段との間に
配置され、前方大角散乱光を検出する前方大角散乱光検
出手段と、 上記前方小角、前方大角及び側方散乱光検出手段から出
力される、前方小角散乱光、側方散乱光及び前方大角散
乱光に対応する検出信号を入力して夫々の散乱光強度デ
ータを算定して記憶する記憶手段と、上記前方小角、前
方大角及び側方散乱光強度データから前方小角散乱光強
度と前方大角散乱光強度の２次元分布データ並びに前方
小角散乱光強度と側方散乱光強度の２次元分布データを
算定する手段と、上記前方小角散乱光強度と前方大角散
乱光強度の２次元分布データから特定の白血球細胞の領
域を決定する手段とを具える分析手段と、 上記算定された前方小角散乱光強度と前方大角散乱光強
度の２次元分布データ、前方小角散乱光強度と側方散乱
光強度の２次元分布データ並びに上記特定された白血球
細胞のデータを上記前方小角散乱光強度と側方散乱光強
度の２次元座標上に表示する表示手段とを具えることを
特徴とする粒子分類装置。1. A particle classification apparatus for irradiating particles with light and detecting scattered light from the particles to classify the particles, comprising: forward scattered light detecting means for detecting forward small-angle scattered light; and the forward scattered light. The front large-angle scattered light detecting means, which is arranged between the detecting means and the side scattered light detecting means and detects the front large-angle scattered light, and the front small angle, the front large angle, and the side scattered light detecting means, the front, Storage means for inputting detection signals corresponding to small-angle scattered light, side-scattered light, and forward large-angle scattered light to calculate and store respective scattered-light intensity data, and the above-mentioned front small-angle, front large-angle, and side-scattered light intensity Means for calculating two-dimensional distribution data of forward small-angle scattered light intensity and forward large-angle scattered light intensity and two-dimensional distribution data of forward small-angle scattered light intensity and side-scattered light intensity, and the above-mentioned forward small-angle scattered light intensity and forward large-angle scattered light Of scattered light intensity Analysis means having means for determining a specific white blood cell region from the two-dimensional distribution data; two-dimensional distribution data of the forward small angle scattered light intensity and the forward large angle scattered light intensity calculated; A display means for displaying the two-dimensional distribution data of the side scattered light intensity and the data of the identified white blood cell on the two-dimensional coordinates of the small front angle scattered light intensity and the side scattered light intensity. Particle sorting device.
細胞としたことを特徴とする請求項１記載の粒子分類装
置。2. The particle classification device according to claim 1, wherein the specific white blood cells are white blood cells having a low content rate.
は好塩基球としたことを特徴とする請求項１又は２記載
の粒子分類装置。3. The particle classification apparatus according to claim 1, wherein the white blood cells having a low content rate are monocytes and / or basophils.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP07689095A JP3350775B2 (en) | 1995-03-31 | 1995-03-31 | Particle classifier |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP07689095A JP3350775B2 (en) | 1995-03-31 | 1995-03-31 | Particle classifier |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH08271509A true JPH08271509A (en) | 1996-10-18 |
| JP3350775B2 JP3350775B2 (en) | 2002-11-25 |
Family
ID=13618245
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP07689095A Expired - Fee Related JP3350775B2 (en) | 1995-03-31 | 1995-03-31 | Particle classifier |
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| Country | Link |
|---|---|
| JP (1) | JP3350775B2 (en) |
Cited By (8)
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| US6084670A (en) * | 1997-03-11 | 2000-07-04 | Nihon Kohden Corporation | Particle analyzer and composite lens formed by integrally joining plural lens elements of different focal points |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US7092078B2 (en) | 2003-03-31 | 2006-08-15 | Nihon Kohden Corporation | Flow cytometer for classifying leukocytes and method for determining detection angle range of the same |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6084670A (en) * | 1997-03-11 | 2000-07-04 | Nihon Kohden Corporation | Particle analyzer and composite lens formed by integrally joining plural lens elements of different focal points |
| US6157500A (en) * | 1997-03-11 | 2000-12-05 | Nihon Kohden Corporation | Particle analyzer and composite lens formed by integrally joining plural lens elements of different focal points |
| US6409141B1 (en) | 1997-03-11 | 2002-06-25 | Nihon Kohden Corporation | Particle analyzer and composite lens formed by integrally joining plural lens elements of different focal points |
| US7130046B2 (en) * | 2004-09-27 | 2006-10-31 | Honeywell International Inc. | Data frame selection for cytometer analysis |
| JP2006230333A (en) * | 2005-02-28 | 2006-09-07 | Hitachi Medical Corp | Flow site meter, method for analyzing cell, cell-analyzing method, method for setting sensitivity of fluorescent light detector and method for setting standard gate in positive rate-judging method |
| JP4649231B2 (en) * | 2005-02-28 | 2011-03-09 | 株式会社カネカ | Flow cytometer, cell analysis method, cell analysis program, sensitivity setting method of fluorescence detector, and reference gate setting method in positive rate determination method |
| WO2014084930A1 (en) * | 2012-11-30 | 2014-06-05 | Beckman Coulter, Inc. | Tuberculosis screening using cpd data |
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