JPH08271509A - Particle sorter - Google Patents

Particle sorter

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JPH08271509A
JPH08271509A JP7689095A JP7689095A JPH08271509A JP H08271509 A JPH08271509 A JP H08271509A JP 7689095 A JP7689095 A JP 7689095A JP 7689095 A JP7689095 A JP 7689095A JP H08271509 A JPH08271509 A JP H08271509A
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scattered light
forward
light intensity
angle
means
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JP3350775B2 (en )
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Yutaka Nagai
Katsuhiro Tsuchiya
勝寛 土屋
豊 永井
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Nippon Koden Corp
日本光電工業株式会社
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Abstract

PURPOSE: To sort respective cells, contained in white blood cells at low content, accurately by detecting them optically.
CONSTITUTION: The particle sorter is provided with an analyzer 10 comprising means 8, 9 for detecting forward small angle scattering light, means 13a, 14a, 15a for detecting forward large angle scattering light interposed between the means 8, 9 and side scattering light detection means 4, 5, 6, means for calculating each scattering light intensity data based on the received detection signal and storing the calculated data, means for calculating a two-dimensional distribution data from the scattering light intensity data, and means for determining the region of a specific white blood cell based on the two-dimensional distribution data of forward small angle scattering light intensity and forward large angle scattering light intensity. The particle sorter further comprises a display 11 for presenting the calculated two-dimensional distribution data of forward small angle scattering light intensity and forward large angle scattering light intensity, the two-dimensional distribution data of forward small angle scattering light intensity and side scattering light intensity, and the data of the specified white blood cell on the two-dimensional coordinate of forward small angle scattering light intensity and side scattering light intensity.
COPYRIGHT: (C)1996,JPO

Description

【発明の詳細な説明】 DETAILED DESCRIPTION OF THE INVENTION

【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。 The present invention relates to relates to, for example, particle classification device for classifying by irradiating light to the particles of white blood cells.

【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。 In general, white blood cells, lymphocytes, monocytes, neutrophils, eosinophils, composed of various cells such as basophils. 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。 These consist each cell nucleus and cytoplasm, each cell, the size, the shape is different, the size of fine particles such as granules contained in the cytoplasm, shape and quantity and the like are known to be different, respectively. 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。 It has been found that the following with respect to granules or particles contained in the content and each cell in each cell. リンパ球(25〜45%);顆粒がほとんどない。 Lymphocytes (25% to 45%); there is little granules. 単球(4〜7%) ;0.1μm以下の顆粒が均一に極めて多数存在する。 Monocytes (4~7%); 0.1μm following granules uniformly present a very large number. 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。 Neutrophils (45-61%); glycogen particles 0.05~0.2μm there are many throughout the cell. 好酸球(1〜5%) ;0.5〜1.0μmの特殊顆粒が多数存在する。 Eosinophils (1~5%); 0.5~1.0μm of specific granules there are many. 好塩基球(0〜1%) ;0.2〜1.0μmの顆粒が多数存在する。 Basophils (0~1%); 0.2~1.0μm granules there are many. ( )内の数値は、各細胞の含有率を示す。 Values ​​in brackets indicate the content of each cell. 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。 Since each of these cells is increased or decreased by the disease, based on information obtained from the granules or particles in these cells, the determination and diagnosis of disease is performed by detecting the state of each cell, the classification of white blood cells Clinical examination in the table it is useful.

【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, these white blood cells is irradiated with a laser beam, cell analysis to detect the two directions of light of the side scattered light representing the granules characteristic of the forward scattered light and the cells or the like which reflects the size of the cell analysis apparatus is widely used in general.

【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。 [0004] Within each cell, as compared to the wavelength of the laser light used, because it contains small granules or particles from larger granules or particles as described above, the scattered light scattered when laser beam irradiation there are directional as shown below. 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 If the size of the granules or particles are large relative to the wavelength; if the magnitude of the forward and side scatter granules or particles is small with respect to wavelength; when the magnitude of the forward scattered light granules or particles of comparable with the wavelength ; front, 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)。 [0005] By vary the size of the granules or particles contained in white blood cells, in the analysis by only conventional forward and side scattered light as described above, since necessarily the cells can not be clearly separated identification, applicant has It proposed a particle sorting device according to the backscattered light in addition to forward and side scatter (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。 [0006] Figure 5 is a schematic view of a particle classifying apparatus present applicant has proposed. 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。 5, flow through the collimating lens 12 having a laser light source 1 lenses 2 and 3 of the laser beam for focusing the illumination light, the mirror 13 and similar holes for reflecting backscattered light having a hole in its central portion It is applied to the chamber 4. 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。 Laser light reflected by the white blood cells contained in the sample liquid in the flow chamber 4 is scattered by such transmitted or diffraction, scattered light is incident on the detector 9 via a forward scattered light focusing lens 8, into an electric signal output is converted. 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。 The forward scattered light converging lens 8 and a detector 9 constitute a forward scattered light detector.

【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。 [0007] The flow chamber 4 is constructed as shown schematically in Figure 6A. 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。 The lumen in the funnel-shaped container 4a, the outer peripheral portion sheath liquid along the (sheath fluid) Sw flows, flows the sample liquid F containing white blood cells to be analyzed the central portion. 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。 In this case, the sheath liquid Sw and the sample liquid F is adapted not to mix the difference in flow rate in accordance with the principles of Reynolds. 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。 Sample liquid F is passing through the narrow tube portion 4b, a laser beam is irradiated.

【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、 [0008] Figure 6B shows a detail of the laser light narrow tube portion 4b is incident on white blood cells C of the sample solution in F,
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。 Here the laser light is scattered by the white blood cells C, the forward scattered light described above, side scattered light and backscattered light is 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。 Further, the scattered light scattered in the perpendicular direction in the fluidized chamber 4 is detected by the configured side scatter light detection means at the side scattered light focusing lens 5 and the detector 6, converted into an electric signal which is to be 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。 Furthermore, backward scattered light scattered in the incident direction opposite to the direction of the laser beam in a fluidized chamber 4, the mirror 13, the backscattered light detecting means composed of backscattered light converging lens 14 and a detector 15 It is detected, and output is converted into an electric signal.

【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。 [0011] forward is detected by the detectors 9,6 and 15, the detection signal corresponding to the side and back-scattered light is stored is input to the analyzer 10, the display device is calculated scattered light intensity of each is is output to the 11. 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。 Display device 11 displays the front, the side and rear scattering intensity data on a two-dimensional coordinates on the screen. 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。 Then, it is possible to perform forward, sideways, by displaying a scattergram by combining the scattered light intensity of the rear on the two-dimensional coordinates, a classification from the distribution of the cells contained in the leukocytes.

【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。 [0012] Thus, the conventional particle sorting apparatus described above, the forward scattered light scattered from the white blood cells, to perform the classification of white blood cells containing small granules or particles by detecting the side scattered light and backscattered light in that it was.

【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。 [SUMMARY OF THE INVENTION However, the front white blood cells containing small granules or particles may be carried out a certain cell classification by detecting the side and back-scattered light, monocytes and basophils low cell of content, such as spheres was difficult to clearly identified or classified buried high content lymphocytes and neutrophils. 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。 Accordingly, the present invention has been made in view of the above problems, and an object thereof is to provide a particle classifier that with few relatively content cells can be accurately classified.

【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、 Particle sorting device according to claim 1 of the present invention In order to achieve the above object, according to the light irradiated to the particles in the particle classifying apparatus performs particles classification by detecting the scattered light from the particles, forward a forward scattered light detector for detecting the small-angle scattered light,
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、 Is disposed between the forward scattered light detecting means and the side scattered light detecting means, the forward high-angle scattered light detecting means for detecting a forward high angle scattered light, is output from the front small-angle, forward large angle and side scatter light detection means , forward small-angle scattered light, and storage means for calculating and storing the scattered light intensity data by entering respectively a detection signal corresponding to the side scattered light and forward high angle scattered light, forward small-angle, forward large angle and side scatter means for calculating a two-dimensional distribution data of the two-dimensional distribution data and forward small-angle scattered light intensity and side scattered light intensity of the forward small-angle scattered light intensity and the forward high-angle scattered light intensity from the intensity data, forward small-angle scattered light intensity and a forward large angle 2D the distribution data and the analyzing means comprising a means for determining the area of ​​a particular white blood cell, calculated by the forward small-angle scattered light intensity and the forward high-angle two-dimensional distribution data of the scattered light intensity of the scattered light intensity,
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。 Is constituted by a display means for displaying the data of the two-dimensional distribution data and the particular leukocyte cell of the forward small-angle scattered light intensity and side scattered light intensity on the two-dimensional coordinates of the forward small-angle scattered light intensity and side scattered light intensity that.

【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。 [0015] The invention according to claim 2 is the particle sorting apparatus according to claim 1, characterized in that a small white blood cells with content specific white blood cells.

【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。 [0016] The invention according to claim 3 is the particle sorting apparatus according to claim 1 or 2, characterized by being less white blood cells with content and monocyte and / or basophil.

【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。 [Action] In the invention according to claim 1, when performing classification by irradiating a laser beam to white blood cells, to detect the forward small-angle scattered light by forward scattering light detecting means, the forward high-angle scattered light by the forward high angle scattered light detecting means detecting side scattered light by the side scattered light detection means detects the. 分析手段により、側方、 The analysis means, the side,
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。 Output from the front and forward high angle scattered light detecting means, the forward small-angle, stores the scattered light intensity data of people each lateral and forward high-angle, two-dimensional distribution data and forward of the forward small-angle scattered light intensity and the forward high-angle scattered light intensity calculated two-dimensional distribution data of the small-angle scattered light intensity and side scattered light intensity, forward small-angle and side to determine the area of ​​specific white blood cells from the two-dimensional distribution data of the forward small-angle scattered light intensity and the forward high-angle scattered light intensity and outputs as a two-dimensional distribution data of the rectangular scattered light intensity. 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、 Two-dimensional distribution data that was calculated forward small-angle scattered light intensity and the forward high-angle scattered light intensity,
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、 The scattered light intensity data of the two-dimensional distribution data and the particular leukocyte cell of the forward small-angle scattered light intensity and side scattered light intensity,
表示手段の画面上に2次元座標上に表示する。 Display on the two-dimensional coordinates on the screen of the display unit.

【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。 [0018] In the invention according to claim 2, selects the smaller white blood cells of content as a specific leukocyte cell.

【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。 [0019] In the invention according to claim 3, selects monocytes or basophils or both as small white blood cells with 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。 EXAMPLES Referring to the drawings, a description will be given of an embodiment of a particle sorting apparatus of the present invention. 図1は、本発明の実施例の構成を示すブロック図である。 Figure 1 is a block diagram showing a configuration of an embodiment of the present invention. 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。 1, parts corresponding to those in FIG. 5 are denoted 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。 [0021] The present example has a side-scattered light detecting means comprising backward scattered light detection lens 5 and the detector 6 shown in FIG. 5, the hole in its central portion, a mirror 13 for reflecting backscattered light, swapping the positions of the backscattered light detecting means comprising backward scattered light detection lens 14 and the detector 15, using the back-scattered light detecting means as a forward high angle scattered light detecting means, and the forward small-angle detection means of the forward scattered light detector those configured for use as a. 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。 Other configurations are the same as the conventional example shown in FIG. 5, description will not be repeated.

【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。 [0022] That is, has a hole in the center, a mirror 13a for reflecting forward large angle scattered light, forward high-angle scattered light focusing lens 14a and a forward high angle scattered light for focusing the forward high angle scattered light reflected by the mirror 13a composed forward high angle scattered light detecting means by a detector 15a for converting into an electric signal.

【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。 Next, the operation of the particle sorting apparatus of the present invention described above. 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 Laser light emitted from the laser light source 1 is focused on the laser light focusing lens 2 and 3, white blood cells C that passes through the narrow tube portion 4b of the flow chamber 4 (Fig. 6)
(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。 Per (Figure 6), the scattered light component scattered laterally is input to the detector 6 is detected by the lens 5 detection side scattered light. 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。 From the detector 6, it is outputted as a detection signal corresponding side scattered light component is converted into an electric signal 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。 Further, direct light incident on the front of the flow chamber 4 (the left in the figure) is shielded by the irradiation light stopper 7, the scattered light other than the direct light is collimated by the scattering light collimator lens 12, It is reflected by the mirror 13a. 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。 The reflected scattered light is input to the detector 15a is detected by the forward high angle scattered light detection lens 14a as the forward high angle scattered light component is converted into an electrical signal. 検出器15aからは、 From the detector 15a is
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。 Is output as a detection signal corresponding to the forward high 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。 Furthermore, the forward small-angle scattering light component having passed through the central hole of the mirror 13 is input to the detector 9 is detected by the forward scattered light detection lens 8. 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。 Detector 9 outputs a detection signal corresponding to the forward small-angle scattered light intensity and converts the forward small-angle scattered light component into an electrical signal.

【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。 [0026] Each detector 6,9,15a described above is constituted by a photoelectric conversion element such as a respective photodiode, for example.

【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。 The front small-angle detected by the detector 6,9,15A, detection signal corresponding to the side and forward high angle scattered light is input to the analyzer 10. 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。 Analyzer 10, input forward small-angle scattered light intensity data from the detected signals were, side scattered light intensity data and forward high angle scattered light intensity data calculated to the store, and forward small-angle and forward high angle scattered light intensity 2 dimensional coordinate data, we calculate two-dimensional coordinate data of the forward small-angle scattered light intensity and side scattered light intensity. 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。 Further, the analyzer 10 is calculated by the forward small-angle scattered light intensity data and forward high angle scattered light intensity data, and sequentially outputs the forward small-angle scattered light intensity data and side scattered light intensity data, or an operation unit of the analyzer 10 output to the display device 11 by the operation (not shown), is displayed as the light intensity scattered distribution diagram of the two-dimensional coordinates on the screen (scattergram).

【0028】図2は、上記実施例によるスキャッタグラムを示す。 [0028] Figure 2 shows a scattergram by the examples. 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。 Figure 2A, the abscissa forward high angle scattered light intensity is taken up vertical axis forward small-angle scattered light intensity, for example lymphocytes, monocytes, neutrophils, a scattergram containing basophils.
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。 Further, FIG. 2B, due to the forward small-angle scattered light intensity (ordinate axis) and side scattered light intensity (abscissa) is a scattergram of the leukocyte cells in Figure 2A.

【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。 [0029] In the scattergram of FIG. 2A and 2B, lymphocytes, monocytes, neutrophils, distribution of cells such as basophils appear interspersed, at a certain identification allows a between each cell the case, the identification of small content such as monocytes and basophils 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。 [0030] Generally, monocytes, have immune function, If this is increasing, it is determined that there is tuberculosis, measles, diseases of acute hepatitis and the like, also, basophils, when increased, thyroid function There is suspicion of degradation and chronic myeloid leukemia, are considered to have diseases such as reduction to the hyperthyroidism. 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。 Therefore, as described above, it is important that even a small content separated identify monocytes or basophils or both.

【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。 [0031] In the scattergram of the normal specimen shown in Figures 2A and 2B, for example, for monocytes and basophils are mixed, it is not possible to clearly identify the two, forward small-angle scattered light intensity and forward large angle to display the scattergram of scattered light intensity, as shown in Figure 2C, to determine the region where monocytes and basophils are mixed (region surrounded by a solid line), the analyzer 10, monocytes and basophils and outputs only the display device 11 the scattered light intensity data of the sphere.

【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。 [0032] In this case, by displaying a scattergram of the forward scattered light intensity (ordinate axis) and side scattered light intensity (horizontal axis), as shown in FIG. 2D, the scattered light intensity of monocytes and basophils data can be identified clearly separated. 図2C Figure 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。 Those determined by monocyte region and scattergram of the normal leukocytes lines identify basophils area shown in lines and Figure 2D showing a region shown in, it is automatically designated by the previously stored control program 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. Further, FIG. 3, for example, in MDS patients scattergram basophils increase multi analyte (myelodysplastic syndrome), taken ordinate the forward scattered light intensity either as in FIG 2, the forward high-angle scattered It has a light intensity and a side scattered light intensity and the horizontal axis. 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。 Figure 3A, lymphocytes, monocytes, neutrophils, a scattergram of the forward small-angle scattered light intensity basophils are mixed and forward high angle scattered light intensity, FIG. 3B is a front small-angle scattered light intensity in FIG. 3A is a scattergram by the 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。 [0034] Figure 3C is for the scattergram of forward small-angle scattered light intensity and the forward high-angle scattered light intensity in FIG. 3A, showing a line that specifies the mixed region of monocytes and basophils. 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。 Further, FIG. 3D, from the specified area by FIG. 3C, taking the scattergram of the forward small-angle scattered light intensity and side scattered light intensity scattergram of basophils up multiple samples of the region identified basophils it 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。 [0035] FIG. 4 is, for example, shows a scattergram of monocytes isolated sample, there is a following method, shown as a method for separating monocytes separated analytes. 抗凝固処理した血液と6%(W/ Anticoagulated blood and 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。 Physiological saline containing V) Dextran500 10: 1 mixture, a greenhouse for 40 minutes after standing, collecting plasma layer. 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 This was layered over the NycoPrep (registered trademark) 1.068, and centrifuged for 15 minutes at 600G, NycoPrep
(登録商標名)の上層から単球浮遊液を採取する。 And collecting the monocyte suspension from the upper layer of the (registered trade name). 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。 Monocytes suspension was diluted with physiological saline containing 0.13% (W / V) EDTA and 1% FCS, and centrifuged 7 minutes at 600G, for cleaning. 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。 Adjustment sample is suspended in autologous plasma to remove the platelets through 0.2μm filters, and monocytes isolated specimens.

【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 [0036] Figure 4A, lymphocytes, a scattergram of monocytes, neutrophils, good front basophils are mixed small-angle scattered light intensity (vertical axis) and a forward high-angle scattered light intensity (horizontal axis), FIG. 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。 Is a scattergram by the forward small-angle scattered light intensity in FIG. 4A (vertical axis) and side scattered light intensity (horizontal axis).

【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。 Further, FIG. 4C, the scattergram of the forward small-angle scattered light intensity and the forward high-angle scattered light intensity in FIG. 4A, a line designating a mixed region of monocytes and basophils is shown one. 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. 4D, from the specified area by FIG 4C, takes the scattergram of the forward small-angle scattered light intensity and side scattered light intensity, a scattergram identify regions of monocytes.

【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 in the foregoing, according to the present invention described in claim 1, the forward small-angle scattered light intensity, forward large angle scattered light intensity, by using the scattergram of side-scattered light intensity, the white blood cells various cells can be clearly separated identification.

【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 described in claim 2, it is easy to separate the identification of small white blood cells with content.

【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。 [0040] Furthermore, according to the present invention described in claim 3, there is a monocyte or basophils or advantages both separation identification is clear.

【図面の簡単な説明】 BRIEF DESCRIPTION OF THE DRAWINGS

【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。 1 is a block diagram showing a configuration of an embodiment of a particle sorting apparatus 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。 It shows a scattergram of a normal sample according to an embodiment of FIG. 1. 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。 3 is a diagram showing a scattergram of basophils increase multiple specimens of MDS patients 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。 Shows a scattergram of monocytes separated analytes by the embodiment of FIG. 1;

【図5】従来例の構成を示すブロック図である。 5 is a block diagram showing a conventional configuration.

【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。 6 is a diagram showing the details of the present invention and flow chamber used in the particle classification device of the conventional example.

【符号の説明】 DESCRIPTION OF SYMBOLS

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 1 laser light source and third lenses 4 flow chamber 5 side scatter light detection lens 8 forward scattered light detection lens 6,9,15a detector 7 irradiation light stopper 10 analyzer for irradiating light converging (analyzing means) 11 Display device (display means) 12 scattered light collimator lens 13a mirrors 14a forward high angle scattered light detection lens

Claims (3)

    【特許請求の範囲】 [The claims]
  1. 【請求項1】 粒子に光を照射し、該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、 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 1. A light is irradiated to the particles, the particle classification device for performing the particle classification by detecting the light scattered by the particles, and the forward scattered light detecting means for detecting the forward small-angle scattered light, the forward scattered light is disposed between the detecting means and the side scattered light detecting means, the forward high-angle scattered light detecting means for detecting a forward high angle scattered light, the forward small-angle, are outputted from the front large-angle and side scatter light detection means, forward Small angle scattered light, and storage means for calculating and storing the scattered light intensity data by entering respectively a detection signal corresponding to the side scattered light and forward high angle scattered light, the forward small-angle, forward large angle and side scatter light intensity two-dimensional distribution data and means for calculating the two-dimensional distribution data of the forward small-angle scattered light intensity and side scattered light intensity, the forward small-angle scattered light intensity and the forward large angle of the forward small-angle scattered light intensity from the data and forward high angle scattered light intensity of the scattered light intensity 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。 And analysis means comprising a means for determining a region of specific white blood cells from the two-dimensional distribution data, two-dimensional distribution data of the forward small-angle scattered light intensity is the calculated and the forward high-angle scattered light intensity, and the forward small-angle scattered light intensity and characterized by comprising a display means for displaying the data of the two-dimensional distribution data and the specified white blood cells of the side scattered light intensity on the two-dimensional coordinates of the forward small-angle scattered light intensity and side scattered light intensity particle classification device to.
  2. 【請求項2】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請求項1記載の粒子分類装置。 2. A specific particle classification apparatus according to claim 1, wherein the white blood cells was less white blood cells with content.
  3. 【請求項3】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請求項1又は2記載の粒子分類装置。 3., characterized in that fewer white blood cells of content and monocyte and / or basophil claim 1 or 2 particle classification apparatus according.
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US7130046B2 (en) * 2004-09-27 2006-10-31 Honeywell International Inc. Data frame selection for cytometer analysis
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JP4649231B2 (en) * 2005-02-28 2011-03-09 株式会社カネカ Flow cytometer, a method analysis of cells, cell analysis program, reference gate setting method in the sensitivity setting method and positive rate determination method of the fluorescence detector
WO2014084930A1 (en) * 2012-11-30 2014-06-05 Beckman Coulter, Inc. Tuberculosis screening using cpd data
CN104949910A (en) * 2015-05-29 2015-09-30 广州埃克森生物科技有限公司 5-Part differential hematology analyzer optical system

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