JP2006234559A - Flow site meter - Google Patents

Flow site meter Download PDF

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JP2006234559A
JP2006234559A JP2005049160A JP2005049160A JP2006234559A JP 2006234559 A JP2006234559 A JP 2006234559A JP 2005049160 A JP2005049160 A JP 2005049160A JP 2005049160 A JP2005049160 A JP 2005049160A JP 2006234559 A JP2006234559 A JP 2006234559A
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flow
fine particles
flow path
sample liquid
flow cell
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Masujiro Hisatani
益士郎 久谷
Nariyuki Nakada
成幸 中田
Hiroyoshi Hayashi
弘能 林
Akihide Ito
彰英 伊藤
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Mitsui Engineering and Shipbuilding Co Ltd
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Mitsui Engineering and Shipbuilding Co Ltd
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<P>PROBLEM TO BE SOLVED: To suppress the occurrence of liquid droplets in which two particulates different in kinds are taken and to enhance accuracy of fraction collection. <P>SOLUTION: In this flow site meter equipped with a main body part for individually discriminating particulates in a process for allowing a sample liquid 14, in which a large number of particulates are floated, to flow through a flow cell 20 and the fraction collection means 28 of particulates on the discharge side of the sample liquid of the flow cell 20, a partition means 38 for throttling a flow channel 36 of the sample liquid connecting the flow cell 20 and the fraction collection means 28 is attached to the flow channel 36. A controller 40 controls the operation of the partition means 38 so as to be matched with the passing time of the particulates, which are discriminated in the main body part, through the partition means 38. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

本発明はフローサイトメータに係り、特にサンプル液の排出側に微粒子の分取手段を具備したフローサイトメータに関する。   The present invention relates to a flow cytometer, and more particularly to a flow cytometer provided with a means for collecting fine particles on the discharge side of a sample liquid.

フローサイトメータは、サンプル液を透明なフローセル内に細長く流すことによって、サンプル液中に存在する細胞、微生物、マイクロビーズなどの微粒子をその特性に応じて光学的に識別し、計数するようにした測定解析装置である。また、必要に応じてサンプル液の排出側に分取手段を取り付け、識別した微粒子を分取手段によって分取することもできる。   The flow cytometer is designed to optically identify and count microparticles such as cells, microorganisms, and microbeads that exist in the sample solution by flowing the sample solution into a transparent flow cell. It is a measurement analysis device. Further, if necessary, a sorting means can be attached to the sample liquid discharge side, and the identified fine particles can be sorted by the sorting means.

図４は一般的なフローサイトメータの概念図である。フローチャンバ１０にはインサーションノズル１２が奥深く挿入され、このインサーションノズル１２の下端開口から微粒子を含んだサンプル液１４がフローチャンバ１０内に注入される。フローチャンバ１０にはシース液の供給口１６が接続している。供給口１６から流れ込んだシース液１８がサンプル液１４を包むようにしてフローチャンバ１０の下部に設けたフローセル２０に向けて細長い押し出し流を形成する。この際シース液１８とサンプル液１４の流量を制御することにより、フローセル２０には微粒子が１個ずつ縦に並んだ状態で流れるようにする。   FIG. 4 is a conceptual diagram of a general flow cytometer. An insertion nozzle 12 is inserted deeply into the flow chamber 10, and a sample liquid 14 containing fine particles is injected into the flow chamber 10 from the lower end opening of the insertion nozzle 12. A sheath liquid supply port 16 is connected to the flow chamber 10. The sheath liquid 18 flowing from the supply port 16 wraps the sample liquid 14 and forms an elongated extruded flow toward the flow cell 20 provided at the lower part of the flow chamber 10. At this time, the flow rates of the sheath liquid 18 and the sample liquid 14 are controlled so that the fine particles flow in the flow cell 20 one by one in a vertically aligned state.

フローセル２０の位置にはレーザ光源２２が配置されており、フローセル２０内にレーザ光を照射している。サンプル液１４中に微粒子が存在するとレーザ光が散乱する。また、微粒子に予め蛍光色素を付与しておくとレーザ光の照射によって微粒子が蛍光を発する。レーザ光源２２の対向位置（又は側向位置）には発生した散乱光や蛍光を測定する検出器２４が配置されている。検出器２４で測定した散乱光や蛍光をデータ処理器２６で解析することによってフローセル２０を通過した個々の微粒子の種類を特定するとともに、所定時間当たりにフローセル２０を通過した微粒子をその種類毎にカウントする。   A laser light source 22 is disposed at the position of the flow cell 20 and irradiates the flow cell 20 with laser light. When fine particles are present in the sample liquid 14, the laser light is scattered. In addition, if a fluorescent dye is previously applied to the fine particles, the fine particles emit fluorescence when irradiated with laser light. A detector 24 for measuring the generated scattered light and fluorescence is arranged at a position (or a side position) opposite to the laser light source 22. By analyzing the scattered light and fluorescence measured by the detector 24 with the data processor 26, the type of each fine particle that has passed through the flow cell 20 is specified, and the fine particle that has passed through the flow cell 20 per predetermined time is identified for each type. Count.

フローセル２０のサンプル液の排出側には微粒子の分取手段２８が設けられている。分取手段２８は制御器２９と荷電器３０と一対の偏向板３２ａ，３２ｂと３個の採取容器３４ａ，３４ｂ，３４ｃとによって構成される。超音波振動などによってフローセル２０の下端ノズルから連続的に滴下する液滴は例えば目的の微粒子１個を含む液滴、微粒子を含まない液滴、目的外の微粒子１個を含む液滴の３種類に区分される。したがって、分取手段２８はこの３種類の液滴をそれぞれ別の採取容器３４ａ，３４ｂ，３４ｃに分取する目的で作動する。   Fine particle sorting means 28 is provided on the sample cell discharge side of the flow cell 20. The sorting means 28 includes a controller 29, a charger 30, a pair of deflecting plates 32a and 32b, and three collection containers 34a, 34b, and 34c. The droplets continuously dropped from the lower end nozzle of the flow cell 20 by ultrasonic vibration or the like are, for example, three types of droplets including a target fine particle, a droplet not containing a fine particle, and a droplet containing a non-target fine particle. It is divided into. Therefore, the sorting means 28 operates for the purpose of sorting the three types of droplets into separate collection containers 34a, 34b, 34c.

すなわち、制御器２９はデータ処理器２６からの信号に基いて、荷電器３０の側方位置を落下する液滴３１が上記３種類の区分のいずれであるかを判別し、液滴３１が目的の微粒子１個を含む液滴である時には例えば液滴３１が−帯電するように荷電器３０を作動させる。また、液滴３１が目的外の微粒子１個を含む液滴である時には液滴３１が＋帯電するように荷電器３０を作動させる。また、液滴３１が微粒子を含まない液滴である時には液滴３１が帯電しないように荷電器３０を作動させる。なお、液滴３１が荷電器３０によって−＋に帯電しやすいように、サンプル液を包むシース液としては電解質溶液が使用される。   That is, based on the signal from the data processor 26, the controller 29 determines which of the above three types of the droplet 31 falling on the side position of the charger 30 is the target of the droplet 31. For example, the charger 30 is operated so that the droplet 31 is charged. Further, when the droplet 31 is a droplet including one undesired fine particle, the charger 30 is operated so that the droplet 31 is positively charged. Further, when the droplet 31 is a droplet not containing fine particles, the charger 30 is operated so that the droplet 31 is not charged. An electrolyte solution is used as the sheath liquid that wraps the sample liquid so that the droplet 31 is easily charged to − + by the charger 30.

荷電器３０の下方には一対の偏向板３２ａ，３２ｂが末広がりに配置されている。偏向板３２ａは＋極板であり、偏向板３２ｂは−極板である。したがって、荷電器３０によって−帯電した液滴３１は偏向板３２ａ，３２ｂ間の空間を落下する過程で＋極板である偏向板３２ａ側に引き寄せられ、偏向板３２ａの下方に配置された採取容器３４ａに採取される。一方、荷電器３０によって＋帯電した液滴３１は−極板である偏向板３２ｂ側に引き寄せられ、偏向板３２ｂの下方に配置された採取容器３４ｃに採取される。帯電していない液滴３１は中央の採取容器３４ｂに採取される。   A pair of deflecting plates 32 a and 32 b are arranged below the charger 30 so as to spread outwardly. The deflection plate 32a is a positive electrode plate, and the deflection plate 32b is a negative electrode plate. Therefore, in the process of dropping the space between the deflecting plates 32a and 32b, the droplet 31 charged by the charger 30 is drawn toward the deflecting plate 32a that is the positive electrode plate, and the collection container disposed below the deflecting plate 32a. Taken at 34a. On the other hand, the droplet 31 positively charged by the charger 30 is drawn toward the deflecting plate 32b, which is a negative electrode, and is collected in a collecting container 34c disposed below the deflecting plate 32b. The uncharged droplet 31 is collected in the central collection container 34b.

その結果、目的の微粒子は採取容器３４ａに、目的外の微粒子は採取容器３４ｃに分取される。また、上記と同様の操作によって、例えば第１目的の微粒子を採取容器３４ａに、第２目的の微粒子を採取容器３４ｃに、目的外の微粒子を採取容器３４ｂに分取することも可能である。このようにして、サンプル液中の微粒子を目的に応じて２〜３種類に分取することができる（以上、例えば非特許文献１又は特許文献１参照）。
ＦＣＭの原理入門講座、［online］、ベックマンコールター社，［平成１７年１月２５日検索］、インターネット＜ＵＲＬ：http//www.bc-cytometry.com/FCM/fcmprinciple.html＞ 特表２００３−５１２６０５号公報 As a result, target fine particles are collected in the collection container 34a, and non-target fine particles are collected in the collection container 34c. Further, by the same operation as described above, for example, it is possible to sort the first target fine particles into the collection container 34a, the second target fine particles into the collection container 34c, and the non-target fine particles into the collection container 34b. In this way, the fine particles in the sample liquid can be sorted into two or three types according to the purpose (see, for example, Non-Patent Document 1 or Patent Document 1).
FCM Principles Introduction Course, [online], Beckman Coulter, Inc. [searched on January 25, 2005], Internet <URL: http // www.bc-cytometry.com / FCM / fcmprinciple.html> Special table 2003-512605 gazette

しかしながら、上記従来技術に係るフローサイトメータではサンプル液中の微粒子相互が接近又は接触状態でフローセル２０に流れ込み、そのままフローセル２０の出口流路から排出される場合がある。このため、排出される１つの液滴中に２個の微粒子が取り込まれる場合がある。上記した分取手段２８は１つの液滴を単位として微粒子を分取するものであるから、１つの液滴に種類の異なる２個の微粒子が取り込まれている時には、これらの微粒子を分けることが不可能である。このため、種類の異なる２個の微粒子を取り込んだ液滴が発生すると、分取精度が低下するという問題点があった。   However, in the flow cytometer according to the above prior art, the fine particles in the sample liquid may flow into the flow cell 20 in an approaching or contacting state and may be discharged as they are from the outlet channel of the flow cell 20. For this reason, two fine particles may be taken into one discharged droplet. Since the above-described sorting means 28 separates the fine particles in units of one droplet, when two different types of fine particles are taken into one droplet, these fine particles can be separated. Impossible. For this reason, when the droplet which took in two fine particles from which a kind differs was generated, there was a problem that sorting accuracy fell.

本発明の目的は上記従来技術の問題点を改善し、種類の異なる２個の微粒子を取り込んだ液滴の発生を抑制し、分取精度を向上させることが可能なフローサイトメータを提供することにある。   An object of the present invention is to provide a flow cytometer that can improve the sorting accuracy by improving the sorting problems by improving the problems of the above prior art, suppressing the generation of droplets incorporating two different types of fine particles. It is in.

上記目的を達成するために本発明に係るフローサイトメータは、複数の微粒子が浮遊するサンプル液をフローセル内に流す過程で前記微粒子を個々に識別するようにした本体部と、前記フローセルのサンプル液の排出側に微粒子の分取手段とを具備したフローサイトメータにおいて、前記フローセルと分取手段とを結ぶサンプル液の流路に当該流路を絞る仕切手段を取り付けたこと特徴とする。この構成のフローサイトメータにおいては、前記本体部で識別した微粒子が前記仕切手段を通過する時刻に合わせて仕切手段の作動を制御する制御手段を具備したことが望ましい。また、前記流路には前記仕切手段を上下方向に２段に配設することができる。前記仕切手段による流路の絞り量は流路断面の７０〜８０％とし、残りの２０〜３０％の流路断面ではサンプル液は流れるが、微粒子は通過できないようにする。   In order to achieve the above object, a flow cytometer according to the present invention includes a main body configured to individually identify the fine particles in a process of flowing a sample liquid in which a plurality of fine particles are suspended in the flow cell, and a sample liquid of the flow cell. In the flow cytometer having fine particle sorting means on the discharge side, a partition means for restricting the flow path is attached to the flow path of the sample liquid connecting the flow cell and the sorting means. In the flow cytometer having this configuration, it is preferable that the flow cytometer further includes a control unit that controls the operation of the partition unit in accordance with the time when the particulates identified by the main body pass through the partition unit. Further, the partitioning means can be arranged in two stages in the vertical direction in the flow path. The amount of restriction of the flow path by the partitioning means is 70 to 80% of the cross section of the flow path, and the sample liquid flows through the remaining flow path cross section of 20 to 30%, but the fine particles cannot pass.

本発明によれば、フローセルと分取手段とを結ぶサンプル液の流路に当該流路を絞る仕切手段を取り付けた構成とされている。このため、種類の異なる微粒子が互いに接近・接触状態で仕切手段の位置を通過する際に、仕切手段を作動させることによって微粒子同士を切り離すことができる。このため、流路の下端開口から滴下する液滴には微粒子が１個づつ取り込まれることになり、後段の分取手段での分取精度が向上する。   According to the present invention, the partition means for restricting the flow path is attached to the flow path of the sample liquid connecting the flow cell and the sorting means. For this reason, when different kinds of fine particles pass through the position of the partitioning means in the state of approaching and contacting each other, the fine particles can be separated by operating the partitioning means. For this reason, the fine particles are taken in one by one in the droplets dropped from the lower end opening of the flow path, and the sorting accuracy in the subsequent sorting means is improved.

また、本発明では本体部で識別した微粒子が仕切手段を通過する時刻に合わせて仕切手段の作動を制御する制御手段を具備しているので、微粒子同士の切り離し操作を適正、確実に行うことができる。   Further, in the present invention, since the control means for controlling the operation of the partitioning means in accordance with the time when the microparticles identified by the main body passes through the partitioning means, the operation for separating the microparticles can be performed appropriately and reliably. it can.

図１は本発明に係るフローサイトメータの実施形態を示す系統図である。図１において、図４と同一の符号を付した要素は、図４に示したものと同一の構成、機能を備えており、その説明を省略する。本実施形態に係るフローセル２０のサンプル液の排出側には、フローセル２０と分取手段２８とを結ぶサンプル液の流路３６が設けられており、この流路３６に当該流路を絞る仕切手段３８が取り付けられている。仕切手段３８としては制御性、作動の敏速性の観点から圧電素子によって作動する仕切弁が好ましく採用される。仕切手段３８は本体部であるデータ処理器２６で識別した微粒子が当該仕切手段３８を通過する時刻に合わせて、その作動が制御器４０によって制御される。   FIG. 1 is a system diagram showing an embodiment of a flow cytometer according to the present invention. 1, elements having the same reference numerals as those in FIG. 4 have the same configurations and functions as those shown in FIG. 4, and descriptions thereof are omitted. A flow path 36 for sample liquid that connects the flow cell 20 and the sorting means 28 is provided on the sample liquid discharge side of the flow cell 20 according to the present embodiment, and partitioning means for restricting the flow path to the flow path 36. 38 is attached. As the partition means 38, a gate valve operated by a piezoelectric element is preferably employed from the viewpoint of controllability and quick operation. The operation of the partition unit 38 is controlled by the controller 40 in accordance with the time when the particulates identified by the data processor 26 as the main body pass through the partition unit 38.

図２は仕切手段３８の作動状況を示した説明図である。図２（１）はフローセル２０において種類の異なる微粒子Ｃと微粒子Ｄが接近又は接触状態で検出された状況を示しており、フローセル２０の下流側の流路３６には微粒子Ａと微粒子Ｂが先行して流れている。このような状況でサンプル液を流し続けると微粒子Ａや微粒子Ｂはそれぞれ一つの液滴に取り込まれて問題なく分取されるが、微粒子Ｃと微粒子Ｄは一つの液滴に取り込まれた状態で流路３６の下端開口から滴下する恐れが強い。   FIG. 2 is an explanatory view showing the operating state of the partition means 38. FIG. 2 (1) shows a situation where different types of fine particles C and fine particles D are detected in the flow cell 20 in an approaching or contacting state, and the fine particles A and B are preceded in the flow path 36 on the downstream side of the flow cell 20. It is flowing. If the sample liquid continues to flow in such a situation, the fine particles A and B are each taken in one droplet and sorted without any problem, but the fine particles C and D are taken in one droplet. There is a strong risk of dripping from the lower end opening of the flow path 36.

したがって、図２（１）に示した種類の異なる微粒子Ｃと微粒子Ｄの接近・接触状態をデータ処理器２６が検出すると、データ処理器２６は検出信号を制御器４０に送る。フローセル２０内及び流路３６を流れるサンプル液の流速は一定であるから、フローセル２０内の所定位置で検出された微粒子Ｃと微粒子Ｄが仕切手段３８の取り付け位置を通過するまでの時間は一定である。したがって、制御器４０はデータ処理器２６からの検出信号に基き、微粒子Ｃと微粒子Ｄが仕切手段３８の取り付け位置を通過する時刻に合わせて仕切手段３８が流路３６を絞る方向に仕切手段３８を作動させる。   Therefore, when the data processor 26 detects the approach / contact state between the different kinds of fine particles C and fine particles D shown in FIG. 2 (1), the data processor 26 sends a detection signal to the controller 40. Since the flow rate of the sample liquid flowing in the flow cell 20 and the flow path 36 is constant, the time until the fine particles C and D detected at a predetermined position in the flow cell 20 pass through the attachment position of the partition means 38 is constant. is there. Therefore, the controller 40 is based on the detection signal from the data processor 26, and the partitioning means 38 in such a direction that the partitioning means 38 narrows the flow path 36 in accordance with the time when the particulates C and D pass the attachment position of the partitioning means 38. Is activated.

図２（２）は仕切手段３８が流路３６を絞った直後の状況を示している。すなわち、仕切手段３８をタイミングよく作動させることによって、微粒子Ｃと微粒子Ｄとを上下に切り離すようにして流路３６が絞られる。仕切手段３８による流路３６の絞り量は流路断面積の７０〜８０％とし、残りの２０〜３０％の流路断面ではサンプル液は流れるが、微粒子Ｄは通過できないようにする。その結果、図２（３）に示したように、微粒子Ｄは仕切手段３８の上方に留まるとともに、微粒子Ｃは絞り部から流入したサンプル液に押し流され、微粒子Ｄと微粒子Ｃとの間隔が大きくなる。   FIG. 2 (2) shows a situation immediately after the partitioning means 38 throttles the flow path 36. That is, by operating the partition means 38 with good timing, the flow path 36 is narrowed so that the fine particles C and D are separated vertically. The amount of restriction of the flow path 36 by the partitioning means 38 is 70 to 80% of the cross-sectional area of the flow path, and the sample liquid flows through the remaining 20 to 30% of the cross-section of the flow path, but the fine particles D cannot pass therethrough. As a result, as shown in FIG. 2 (3), the fine particles D remain above the partitioning means 38, and the fine particles C are pushed away by the sample liquid flowing in from the constricted portion, so that the distance between the fine particles D and the fine particles C is large. Become.

そこで、仕切手段３８を開くとサンプル液の流れが回復する。微粒子Ｄと微粒子Ｃとの間隔が大きくなった結果、図２（４）に示したように、微粒子Ｃを含む液滴に微粒子Ｄが同伴することを防止できる。このため、流路３６の下端開口から滴下する液滴には微粒子Ｃや微粒子Ｄが１個づつ取り込まれることになり、後段の分取手段２８での分取精度が向上する。   Therefore, when the partition means 38 is opened, the flow of the sample liquid is recovered. As a result of the increase in the distance between the fine particles D and the fine particles C, it is possible to prevent the fine particles D from accompanying the droplets containing the fine particles C as shown in FIG. For this reason, the fine particles C and the fine particles D are taken in one by one in the droplets dropped from the lower end opening of the flow path 36, and the sorting accuracy in the subsequent sorting means 28 is improved.

なお、フローセル２０において微粒子が適正な間隔を保持して流れているか、又は同種類の微粒子が相互に接近・接触状態である時は、分取操作上の問題が発生しないので、制御器４０は仕切手段３８を格別に作動させる必要はない。   Note that when the fine particles are flowing in the flow cell 20 while maintaining an appropriate interval, or when the same kind of fine particles are close to each other and in contact with each other, there is no problem in the sorting operation. There is no need to actuate the partition means 38 specially.

図３は本発明に係るフローサイトメータの他の実施形態とその作動状況を示した説明図である。本実施形態では流路３６に仕切手段３８Ａ，３８Ｂが所定の間隔で上下２段に配設されている。図３（１）はフローセル２０において種類の異なる３個の微粒子Ｆ，Ｇ，Ｈが接近又は接触状態で検出された状況を示している。このような状況でサンプル液を流し続けると種類の異なる微粒子同士が一つの液滴に取り込まれた状態で流路３６の下端開口から滴下する恐れが強い。   FIG. 3 is an explanatory view showing another embodiment of the flow cytometer according to the present invention and its operating condition. In the present embodiment, partitioning means 38A, 38B are disposed in the flow path 36 in two upper and lower stages at a predetermined interval. FIG. 3 (1) shows a situation in which three different types of fine particles F, G, H are detected in the flow cell 20 in an approaching or contacting state. If the sample liquid is kept flowing in such a situation, there is a strong possibility that different types of fine particles are dropped from the lower end opening of the flow path 36 in a state where the fine particles are taken into one droplet.

したがって、このような状態をデータ処理器２６が検出すると、データ処理器２６は検出信号を制御器４０に送る。すると、制御器４０はデータ処理器２６からの検出信号に基き、微粒子Ｆ，Ｇ，Ｈが第１段の仕切手段３８Ａの取り付け位置を通過する時刻に合わせて流路３６を絞る方向に仕切手段３８Ａを作動させる。   Therefore, when the data processor 26 detects such a state, the data processor 26 sends a detection signal to the controller 40. Then, the controller 40 is based on the detection signal from the data processor 26, and the partition means in the direction of narrowing the flow path 36 in accordance with the time when the fine particles F, G, H pass the mounting position of the first-stage partition means 38A. Activate 38A.

図３（２）は仕切手段３８Ａが流路３６を絞った直後の状況を示している。すなわち、仕切手段３８Ａをタイミングよく作動させることによって、微粒子Ｆを微粒子Ｇ，Ｈから切り離すようにして流路３６が絞られる。その結果、図３（３）に示したように、微粒子Ｇ，Ｈは仕切手段３８Ａの上方に留まるとともに、微粒子Ｆは絞り部から流入したサンプル液に押し流され、微粒子Ｆと微粒子Ｇ，Ｈとの間隔が大きくなる。   FIG. 3 (2) shows a situation immediately after the partitioning means 38 A throttles the flow path 36. That is, by operating the partitioning means 38A with good timing, the flow path 36 is narrowed so as to separate the fine particles F from the fine particles G and H. As a result, as shown in FIG. 3 (3), the fine particles G and H remain above the partitioning means 38A, and the fine particles F are pushed away by the sample liquid flowing in from the constricted portion, and the fine particles F, the fine particles G and H, and The interval of becomes larger.

そこで、仕切手段３８Ａを開くとサンプル液の流れが回復し、図３（４）に示したように、微粒子Ｆと間隔をあけた微粒子Ｇ，Ｈが接近・接触状態で流れる。このような状況でサンプル液を流し続けると、微粒子Ｆは問題ないが、種類の異なる微粒子Ｇ，Ｈ同士が一つの液滴に取り込まれた状態で流路３６の下端開口から滴下する恐れが強い。したがって、制御器４０は微粒子Ｇ，Ｈが第２段の仕切手段３８Ｂの取り付け位置を通過する時刻に合わせて流路３６を絞る方向に仕切手段３８Ｂを作動させる。   Therefore, when the partitioning means 38A is opened, the flow of the sample liquid is restored, and as shown in FIG. 3 (4), the fine particles G and H spaced apart from each other flow in an approaching / contacting state. If the sample liquid continues to flow in such a situation, there is no problem with the fine particles F, but there is a strong possibility that the different types of fine particles G and H will be dropped from the lower end opening of the flow path 36 in a state where they are taken into one droplet. . Therefore, the controller 40 operates the partition unit 38B in the direction of narrowing the flow path 36 in accordance with the time when the fine particles G and H pass the attachment position of the second stage partition unit 38B.

図３（５）は仕切手段３８Ｂが流路３６を絞った直後の状況を示している。すなわち、仕切手段３８Ｂをタイミングよく作動させることによって、微粒子Ｇを微粒子Ｈから切り離すようにして流路３６が絞られる。その結果、図３（６）に示したように、微粒子Ｈは仕切手段３８Ｂの上方に留まるとともに、微粒子Ｇは絞り部から流入したサンプル液に押し流され、微粒子Ｇと微粒子Ｈとの間隔が大きくなる。次に図３（７）に示したように、仕切手段３８を開くとサンプル液の流れが回復し、微粒子Ｇと微粒子Ｈは適正な間隔を有して流路３６を流れることになる。   FIG. 3 (5) shows a situation immediately after the partitioning means 38 </ b> B throttles the flow path 36. That is, the flow path 36 is narrowed so that the fine particles G are separated from the fine particles H by operating the partitioning means 38B with good timing. As a result, as shown in FIG. 3 (6), the fine particles H remain above the partitioning means 38B, and the fine particles G are pushed away by the sample liquid flowing in from the constricted portion, and the interval between the fine particles G and the fine particles H is large. Become. Next, as shown in FIG. 3 (7), when the partition means 38 is opened, the flow of the sample liquid is restored, and the fine particles G and H flow through the flow path 36 with an appropriate interval.

このため、流路３６の下端開口から滴下する液滴にはフローセル２０を通過した時点では接近・接触状態であった３個の微粒子Ｆ，Ｇ，Ｈがそれぞれ間隔をあけて流路３６に流れることになる。したがって、流路３６の出口では微粒子Ｆ，Ｇ，Ｈがそれぞれ１個づつ液滴に取り込まれて滴下することになり、後段の分取手段２８での分取精度が向上する。   For this reason, in the liquid droplet dripping from the lower end opening of the flow path 36, the three fine particles F, G, and H that were in the approach / contact state at the time of passing through the flow cell 20 flow into the flow path 36 at intervals. It will be. Therefore, the fine particles F, G, and H are respectively taken into the droplets and dropped at the outlet of the flow path 36, and the sorting accuracy in the subsequent sorting unit 28 is improved.

本発明に係るフローサイトメータの実施形態を示す系統図である。It is a systematic diagram showing an embodiment of a flow cytometer according to the present invention. 仕切手段３８の作動状況を示した説明図である。It is explanatory drawing which showed the operating condition of the partition means. 本発明に係るフローサイトメータの他の実施形態とその作動状況を示した説明図である。It is explanatory drawing which showed other embodiment of the flow cytometer which concerns on this invention, and its operating condition. 従来技術の一般的なフローサイトメータの概念図である。It is a conceptual diagram of the general flow cytometer of a prior art.

符号の説明Explanation of symbols

１０………フローチャンバ、１２………インサーションノズル、１４………サンプル液、１６………（シース液の）供給口、１８………シース液、２０………フローセル、２２………レーザ光源、２４………検出器、２６………データ処理器、２８………分取手段、３０………荷電器、３２ａ，３２ｂ………偏向板、３４ａ，３４ｂ，３４ｃ………採取容器、３６………流路、３８，３８Ａ，３８Ｂ………仕切手段、４０………制御器。 10 ......... Flow chamber, 12 ......... Insertion nozzle, 14 ......... Sample liquid, 16 ......... Supply port (sheath liquid), 18 ......... Sheath liquid, 20 ......... Flow cell, 22 ... ... Laser light source, 24 ... Detector, 26 ... Data processor, 28 ... Sorting means, 30 ... Charger, 32a, 32b ......... Deflecting plates, 34a, 34b, 34c ... ... collection container, 36 ... flow path, 38, 38A, 38B ... partitioning means, 40 ... controller.

Claims (4)

複数の微粒子が浮遊するサンプル液をフローセル内に流す過程で前記微粒子を個々に識別するようにした本体部と、前記フローセルのサンプル液の排出側に微粒子の分取手段とを具備したフローサイトメータにおいて、前記フローセルと分取手段とを結ぶサンプル液の流路に当該流路を絞る仕切手段を取り付けたこと特徴とするフローサイトメータ。   A flow cytometer comprising: a main body part for individually identifying the fine particles in a process of flowing a sample liquid in which a plurality of fine particles are suspended in the flow cell; and a fine particle sorting means on the sample liquid discharge side of the flow cell The flow cytometer according to claim 1, wherein a partition means for restricting the flow path is attached to the flow path of the sample liquid connecting the flow cell and the sorting means. 前記本体部で識別した微粒子が前記仕切手段を通過する時刻に合わせて仕切手段の作動を制御する制御手段を具備したことを特徴とする請求項１に記載のフローサイトメータ。   The flow cytometer according to claim 1, further comprising a control unit that controls the operation of the partitioning unit in accordance with a time at which the particulates identified by the main body pass through the partitioning unit. 前記流路には前記仕切手段が上下方向に２段に配設されていることを特徴とする請求項１又は請求項２に記載のフローサイトメータ。   The flow cytometer according to claim 1 or 2, wherein the partitioning means is disposed in the flow path in two stages in the vertical direction. 前記仕切手段による流路を絞り量が流路断面の７０〜８０％であり、残りの２０〜３０％の流路断面ではサンプル液は流れるが、前記微粒子は通過できないようにされたことを特徴とする請求項１乃至請求項３のいずれかに記載のフローサイトメータ。   The amount of restriction in the flow path by the partitioning means is 70 to 80% of the cross section of the flow path, and the sample liquid flows in the remaining 20 to 30% of the flow path cross section, but the fine particles cannot be passed. The flow cytometer according to any one of claims 1 to 3.
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