JPH0478938B2 - - Google Patents
Info
- Publication number
- JPH0478938B2 JPH0478938B2 JP61112985A JP11298586A JPH0478938B2 JP H0478938 B2 JPH0478938 B2 JP H0478938B2 JP 61112985 A JP61112985 A JP 61112985A JP 11298586 A JP11298586 A JP 11298586A JP H0478938 B2 JPH0478938 B2 JP H0478938B2
- Authority
- JP
- Japan
- Prior art keywords
- flow cell
- flow
- reflection
- refractive index
- 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.)
- Expired - Lifetime
Links
- 239000002245 particle Substances 0.000 claims description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 239000000463 material Substances 0.000 claims 1
- 230000003287 optical effect Effects 0.000 description 7
- 230000004888 barrier function Effects 0.000 description 5
- 238000003384 imaging method Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 239000012491 analyte Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1456—Electro-optical investigation, e.g. flow cytometers without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
- G01N15/1459—Electro-optical investigation, e.g. flow cytometers without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Electro-optical investigation, e.g. flow cytometers
- G01N15/1434—Electro-optical investigation, e.g. flow cytometers using an analyser being characterised by its optical arrangement
- G01N2015/144—Imaging characterised by its optical setup
- G01N2015/1443—Auxiliary imaging
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6482—Sample cells, cuvettes
Description
【発明の詳細な説明】
［産業上の利用分野］
本発明は、表面に光学的処理を施した粒子解析
装置用フローセルに関するものである。DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a flow cell for a particle analysis device whose surface is optically treated.
［従来の技術］
フローセルメータ等に用いられる従来の粒子解
析装置用フローセルでは、フローセルの中央部の
例えば200μｍ×200μｍの微少な断面を有する流
通部内を、シース液に包まれて通過する検体粒子
に照射光を照射し、その結果生ずる前方及び側方
散乱光により、検体粒子の形状・大きさ・屈折率
等の粒子的性質を得ることが可能である。[Prior Art] In a conventional flow cell for a particle analyzer used in a flow cell meter, etc., sample particles passing through a flow section surrounded by a sheath liquid and having a minute cross section of, for example, 200 μm x 200 μm, are located in the center of the flow cell. It is possible to obtain particle properties such as shape, size, refractive index, etc. of sample particles by irradiating the irradiation light and using the resulting forward and side scattered light.
一般に、レーザー光はAr+レーザーの488nｍの
波長光が使用されており、蛍光は可視光域全体に
渡つている。レーザー光源からのレーザー光Ｌが
フローセル１内に入射した場合に、第２図に示す
フローセル１の入射方向のａ側、ｂ側の面でフレ
ネル反射して数％の反射が生ずる。従つて、ａ
側、ｂ側の面での反射は複数回の反射を繰り返し
た後に、集光レンズに入射し光検出器へ到達し、
この光は検体粒子が存在しない場合でも、光検出
器の出力信号の直流成分として現われることにな
る。この直流成分は検出のダイナミツクレンジを
低下させるという欠点があると共に、レーザー光
の雑音による光信号の変動も出力として大きくな
るので、検体粒子の検出精度に与える影響も大き
い。 Generally, Ar + laser light with a wavelength of 488 nm is used as the laser light, and the fluorescence spans the entire visible light range. When laser light L from a laser light source enters the flow cell 1, Fresnel reflection occurs on the a side and b side surfaces of the flow cell 1 in the direction of incidence shown in FIG. 2, resulting in reflection of several percent. Therefore, a
After the reflection from the side and b side surfaces repeats multiple reflections, it enters the condenser lens and reaches the photodetector.
This light will appear as the DC component of the photodetector output signal even in the absence of analyte particles. This DC component has the disadvantage of reducing the dynamic range of detection, and also increases the fluctuation of the optical signal due to noise in the laser beam as an output, which has a large effect on the detection accuracy of the sample particles.
そこで、既に特開昭61−66947号公報に開示さ
れているように、本出願人はフローセルの表面に
反射防止膜を施して有害光を減少する発明を提案
した。 Therefore, as already disclosed in Japanese Unexamined Patent Publication No. 61-66947, the present applicant proposed an invention in which an antireflection film is applied to the surface of a flow cell to reduce harmful light.
しかしながら、実際にはフローセル１の流通部
１ａの内面においても、表面ほどではないにして
も反射が生じ、その影響を無視することはできな
い。 However, in reality, reflection also occurs on the inner surface of the flow section 1a of the flow cell 1, although not as much as on the surface, and its influence cannot be ignored.
［発明の目的］
本発明の目的は、フローセルの内面反射による
有害光を減少する粒子解析装置用フローセルを提
供することにある。[Object of the Invention] An object of the present invention is to provide a flow cell for a particle analyzer that reduces harmful light due to internal reflection of the flow cell.
［発明の概要］
上述の目的を達成するための本発明の要旨は、
フローセル内の流通部を流れる検体粒子に光を照
射し、検体粒子から得られる散乱光又は蛍光を測
光する装置に使用されるフローセルであつて、前
記流通部の内面に反射防止膜を施したことを特徴
とする粒子解析装置用フローセルである。[Summary of the invention] The gist of the present invention for achieving the above object is as follows:
A flow cell used in a device that irradiates light onto sample particles flowing through a flow section in the flow cell and measures scattered light or fluorescence obtained from the sample particles, the flow cell having an anti-reflection coating applied to the inner surface of the flow section. This is a flow cell for a particle analysis device, which is characterized by:
［発明の実施例］
本発明を図示の実施例に基づいて詳細に説明す
る。[Embodiments of the Invention] The present invention will be described in detail based on illustrated embodiments.
第１図は光学系の構成図であり、フローセル１
の流通部１ａ内を高速層流となつたシース液に包
まれて、液体力学的焦点合わせが行われたサンプ
ル液が通過し、この流れと直交する方向にレーザ
ー光源２が配置されている。このレーザー光源２
から照射されたレーザー光Ｌを流通部１ａに導く
ために、光軸０１上に結像レンズ３が配置されて
おり、更に検体粒子から得られる前方散乱光を測
定するために、フローセル１を挟んで結像レンズ
３と反対側の光軸０１上にストツパ４、集光レン
ズ５、光検出器６が順次に配列されている。ま
た、検体粒子の流れの方向と光軸０１にそれぞれ
直交する方向である光軸０２上に、フローセル１
側から集光レンズ７、コリメートレンズ８、波長
選択特性を有するダイクロイツクミラー９及び反
射ミラー１０が順次に配列されている。そして、
ダイクロイツクミラー９の反射方向にはバリアフ
イルタ１１、集光レンズ１２、光検出器１３が、
反射ミラー１０の反射方向にはバリアフイルタ１
４、集光レンズ１５、光検出器１６が配列されて
いる。 Figure 1 is a configuration diagram of the optical system, and the flow cell 1
A sample liquid that has been hydrodynamically focused passes through the flow section 1a surrounded by the sheath liquid in a high-speed laminar flow, and a laser light source 2 is arranged in a direction perpendicular to this flow. This laser light source 2
An imaging lens 3 is arranged on the optical axis 01 in order to guide the laser beam L irradiated from the analyte to the flow section 1a, and an imaging lens 3 is arranged on the optical axis 01 with the flow cell 1 in between to measure the forward scattered light obtained from the sample particles. A stopper 4, a condensing lens 5, and a photodetector 6 are sequentially arranged on the optical axis 01 on the opposite side from the imaging lens 3. In addition, a flow cell 1 is placed on the optical axis 02, which is a direction perpendicular to the flow direction of the sample particles and the optical axis 01.
A condensing lens 7, a collimating lens 8, a dichroic mirror 9 having wavelength selection characteristics, and a reflecting mirror 10 are arranged in this order from the side. and,
In the reflection direction of the dichroic mirror 9, a barrier filter 11, a condensing lens 12, and a photodetector 13 are installed.
A barrier filter 1 is provided in the reflection direction of the reflection mirror 10.
4. A condenser lens 15 and a photodetector 16 are arranged.
第１図において、レーザー光源２からのレーザ
ー光Ｌは結像レンズ３でフローセル１の流通部１
ａ付近に結像される。フローセル１を流れるサン
プル液の中の検体粒子に当つたレーザー光Ｌによ
る散乱光のうち、サイズを調べるのに適した前方
散乱光は集光レンズ５により光検出器６に集めら
れる。なお、この場合にストツパ４はレーザー光
Ｌの直接光をカツトする役割を果している。 In FIG. 1, a laser beam L from a laser light source 2 passes through an imaging lens 3 to a flow section 1 of a flow cell 1.
The image is formed near a. Among the scattered light caused by the laser beam L hitting the sample particles in the sample liquid flowing through the flow cell 1, forward scattered light suitable for examining the size is collected by the condenser lens 5 onto the photodetector 6. In this case, the stopper 4 plays a role of cutting off the direct light of the laser light L.
また、検体粒子が蛍光標識されている場合に
は、90゜方向に設けられている集光レンズ７、コ
リメートレンズ８、ダイクロイツクミラー９、バ
リアフイルタ１１，１４、集光レンズ１２，１５
を用いて、複数チヤンネルの蛍光測定が可能であ
る。即ち、光検出器１３，１６からの信号を解析
することにより、蛍光標識粒子の分析分類が可能
となる。このとき、バリアフイルタ１１，１４は
それぞれ必要な波長域の光のみを通す特性を有し
ている。 In addition, when the sample particles are fluorescently labeled, the condenser lens 7, collimator lens 8, dichroic mirror 9, barrier filters 11, 14, condenser lenses 12, 15 provided in the 90° direction
Multi-channel fluorescence measurement is possible using this method. That is, by analyzing the signals from the photodetectors 13 and 16, the fluorescent labeled particles can be analyzed and classified. At this time, each of the barrier filters 11 and 14 has a characteristic of passing only light in a necessary wavelength range.
ここで、流通部１ａの内面には反射防止膜がコ
ーテイングされている。この反射防止膜はフロー
セル１を構成する例えばガラスの屈折率とサンプ
ル液の主成分である水の屈折率との中間の屈折率
を有することが必要である。また内面へのコーテ
イングは、コーテイングした複数個のガラスブロ
ツクを組立てることによりフローセル１を製作す
ればよい。 Here, the inner surface of the flow section 1a is coated with an antireflection film. This anti-reflection film needs to have a refractive index that is intermediate between the refractive index of, for example, glass constituting the flow cell 1 and the refractive index of water, which is the main component of the sample liquid. Further, the inner surface may be coated by manufacturing the flow cell 1 by assembling a plurality of coated glass blocks.
この内面へのコーテイングと共に、フローセル
１のａ側、ｂ側の面に特定波長域に対し反射防止
効果の高い反射防止膜を施すことによつて、入出
射面におけるフレネル反射を小さくし、有害光を
カツトすることが可能となる。また、ｃ側の面は
蛍光測定用の光が出射して行く方向であり、ここ
には使用波長域に渡つて反射の少ない反射防止膜
をコーテイングすることにより、集光の効率を高
めることができる。同様に、ｄ側の面においても
反射防止膜により反射率を低下させることが好ま
しい。 In addition to this coating on the inner surface, an anti-reflection film that has a high anti-reflection effect in a specific wavelength range is applied to the a-side and b-side surfaces of the flow cell 1, thereby reducing Fresnel reflection on the input and output surfaces and blocking harmful light. It becomes possible to cut. In addition, the c-side surface is the direction in which the light for fluorescence measurement is emitted, and by coating this with an anti-reflection film that has low reflection over the wavelength range used, it is possible to increase the efficiency of light collection. can. Similarly, it is preferable to reduce the reflectance on the d-side surface with an antireflection film.
［発明の効果］
以上説明したように本発明に係る粒子解析装置
用フローセルは、フローセルに使用レーザー光の
波長の反射を特に低下させる反射防止膜をフロー
セルの内面にコーテイングすることにより有害光
が低減できる。[Effects of the Invention] As explained above, the flow cell for a particle analyzer according to the present invention reduces harmful light by coating the inner surface of the flow cell with an anti-reflection film that particularly reduces the reflection of the wavelength of the laser light used in the flow cell. can.
図面は本発明に係る粒子解析装置用フローセル
の実施例を示し、第１図はその構成図、第２図は
フローセルの斜視図である。
符号１はフローセル、１ａは流通部、２はレー
ザー光源、３は結像レンズ、４はストツパ、５，
７，１２，１５は集光レンズ、６，１３，１６は
光検出器、９はダイクロイツクミラー、１０は反
射ミラー、１１，１４はバリアフイルタである。
The drawings show an embodiment of a flow cell for a particle analyzer according to the present invention, and FIG. 1 is a configuration diagram thereof, and FIG. 2 is a perspective view of the flow cell. 1 is a flow cell, 1a is a flow section, 2 is a laser light source, 3 is an imaging lens, 4 is a stopper, 5,
7, 12, and 15 are condenser lenses; 6, 13, and 16 are photodetectors; 9 is a dichroic mirror; 10 is a reflecting mirror; and 11, 14 are barrier filters.
Claims (1)
を照射し、検体粒子から得られる散乱光又は蛍光
を測光する装置に使用されるフローセルであつ
て、前記流通部の内面に反射防止膜を施したこと
を特徴とする粒子解析装置用フローセル。 ２ 前記反射防止膜は前記フローセルの材質の屈
折率と水の屈折率との中間の屈折率を有するもの
とした特許請求の範囲第１項に記載の粒子解析装
置用フローセル。 ３ 前記フローセルの外面に反射防止膜を施した
特許請求の範囲第１項に記載の粒子解析装置用フ
ローセル。[Scope of Claims] 1. A flow cell used in a device that irradiates light onto sample particles flowing through a flow path in the flow cell and measures scattered light or fluorescence obtained from the sample particles, wherein the flow cell has an inner surface of the flow section. A flow cell for particle analysis equipment characterized by being coated with an anti-reflection film. 2. The flow cell for a particle analyzer according to claim 1, wherein the antireflection film has a refractive index intermediate between the refractive index of the material of the flow cell and the refractive index of water. 3. The flow cell for a particle analyzer according to claim 1, wherein an antireflection film is applied to the outer surface of the flow cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61112985A JPS62269042A (en) | 1986-05-17 | 1986-05-17 | Flow cell for particle analyser |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61112985A JPS62269042A (en) | 1986-05-17 | 1986-05-17 | Flow cell for particle analyser |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS62269042A JPS62269042A (en) | 1987-11-21 |
| JPH0478938B2 true JPH0478938B2 (en) | 1992-12-14 |
Family
ID=14600521
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61112985A Granted JPS62269042A (en) | 1986-05-17 | 1986-05-17 | Flow cell for particle analyser |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62269042A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013250203A (en) * | 2012-06-01 | 2013-12-12 | Denso Corp | Fluid component sensor |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0592701U (en) * | 1992-04-22 | 1993-12-17 | 株式会社キング製作所 | Spectroscopic cell |
| DE102012108158B4 (en) * | 2012-09-03 | 2016-03-17 | Johann Wolfgang Goethe-Universität | Capillary cell, assembly and method for receiving, positioning and examining a microscopic sample |
-
1986
- 1986-05-17 JP JP61112985A patent/JPS62269042A/en active Granted
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013250203A (en) * | 2012-06-01 | 2013-12-12 | Denso Corp | Fluid component sensor |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS62269042A (en) | 1987-11-21 |
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