JPS5913697B2 - Cancer diagnostic equipment using a fluorescent polarization photometric microscope - Google Patents

Cancer diagnostic equipment using a fluorescent polarization photometric microscope

Info

Publication number
JPS5913697B2
JPS5913697B2 JP52036200A JP3620077A JPS5913697B2 JP S5913697 B2 JPS5913697 B2 JP S5913697B2 JP 52036200 A JP52036200 A JP 52036200A JP 3620077 A JP3620077 A JP 3620077A JP S5913697 B2 JPS5913697 B2 JP S5913697B2
Authority
JP
Japan
Prior art keywords
light
cancer
fluorescence
polarization
cancer diagnostic
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
Application number
JP52036200A
Other languages
Japanese (ja)
Other versions
JPS53122489A (en
Inventor
一郎 沢村
保博 中村
健 米窪
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Olympus Corp
Original Assignee
Olympus Optical Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Olympus Optical Co Ltd filed Critical Olympus Optical Co Ltd
Priority to JP52036200A priority Critical patent/JPS5913697B2/en
Priority to DE19782813317 priority patent/DE2813317A1/en
Publication of JPS53122489A publication Critical patent/JPS53122489A/en
Publication of JPS5913697B2 publication Critical patent/JPS5913697B2/en
Expired legal-status Critical Current

Links

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6445Measuring fluorescence polarisation
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/0059Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/41Detecting, measuring or recording for evaluating the immune or lymphatic systems
    • A61B5/414Evaluating particular organs or parts of the immune or lymphatic systems
    • A61B5/415Evaluating particular organs or parts of the immune or lymphatic systems the glands, e.g. tonsils, adenoids or thymus
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/21Polarisation-affecting properties
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/645Specially adapted constructive features of fluorimeters
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/0004Microscopes specially adapted for specific applications

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Pathology (AREA)
  • Immunology (AREA)
  • Chemical & Material Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Analytical Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Engineering & Computer Science (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Surgery (AREA)
  • Animal Behavior & Ethology (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Medical Informatics (AREA)
  • Biomedical Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Optics & Photonics (AREA)
  • Endocrinology (AREA)
  • Vascular Medicine (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Microscoopes, Condenser (AREA)

Description

【発明の詳細な説明】 本発明は蛍光偏光測光顕微鏡を用いたガン診断装置に関
するものである。
DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a cancer diagnostic device using a fluorescence polarization photometric microscope.

最近被検者より採取した血液にガン細胞より取出したた
ん白を混ぜたものをFDA(Fluoresceind
iacetate)にて染色し、これに偏光した励起光
をあて蛍光を発せしめ、この蛍光の偏光度を測定するこ
とによつてガンの診断を行なう方法が発見された。
The FDA (Fluoresceind.
A method has been discovered for diagnosing cancer by staining with acetate, emitting fluorescence by exposing it to polarized excitation light, and measuring the degree of polarization of this fluorescence.

それはガン患者のリンパ球はガン抗原(ガンたん白)に
よつて特異的に反応し、リンパ球がこの抗原ありいはこ
れと類似の物質に接すると何らかの刺激を受ける。そし
てこのリンパ球の刺激によつて起される細胞質の変化が
極めて短時間に細胞の蛍光偏光という物理的性質の変化
とし5 て検出されることによる。したがつて、前述の
ように被検者の血液からリンパ球だけを分離しこれにガ
ンたん白を混せたものをFDAで染色しこれに偏光した
励起光をあてた時に発する蛍光のうち励起光と同じ振動
方向の成分をI”’これと垂直の10振動方向をもつ成
分を11とすると、I’ にQI、P■−(Qは定数) I″+QI、 で表わされる蛍光偏光度Pを求めれば良い。
The lymphocytes of cancer patients react specifically to cancer antigens (cancer proteins), and when the lymphocytes come into contact with this antigen or a substance similar to it, they are stimulated in some way. Changes in the cytoplasm caused by this lymphocyte stimulation are detected in an extremely short time as changes in the physical properties of the cells, such as fluorescence polarization. Therefore, as mentioned above, only lymphocytes are separated from the blood of a subject, mixed with cancer protein, stained with FDA, and when polarized excitation light is applied to the lymphocytes, the excitation of the fluorescence emitted is If the component with the same vibration direction as the light is I"' and the component with 10 vibration directions perpendicular to this is 11, then I' is QI, P - (Q is a constant) I" + QI, the degree of fluorescence polarization P expressed as All you have to do is ask for it.

この蛍光偏光度Pの値は正常人では1.19〜1.59
で15あるのに対し、ガン患者では0.66〜0.86
である。そしてこの方法はガンたん白を変えることによ
り被検者がガン患者の場合、ガンの種類も判定すること
が出来る。本発明は上述の診断方法をもとにして、試料
に20偏光した励起光をあて蛍光を生ぜしめ、この蛍光
の偏光度を電気的に処理することによつてガンであるか
否かの診断を自動的に行ない得るようにしたガン診断装
置を提供するものである。
The value of this fluorescence polarization degree P is 1.19 to 1.59 in normal people.
In cancer patients, the number is 15, whereas in cancer patients it is 0.66 to 0.86.
It is. If the subject is a cancer patient, this method can also determine the type of cancer by changing the cancer protein. The present invention is based on the above-described diagnostic method, in which a sample is exposed to 20 polarized excitation light to generate fluorescence, and the degree of polarization of this fluorescence is electrically processed to diagnose whether or not it is cancer. To provide a cancer diagnostic device that can automatically perform the following.

以下本発明装置の詳細な内容を説明すると、図25にお
いて1は光源、2はコレクターレンズ、3はシャッター
、4はコンデンサーレンズでこれらで一般の観察用透過
照明系を構成する。
The details of the apparatus of the present invention will be described below. In FIG. 25, 1 is a light source, 2 is a collector lens, 3 is a shutter, and 4 is a condenser lens, which constitute a general transmitted illumination system for observation.

5は試料、6は対物レンズである。5 is a sample, and 6 is an objective lens.

又7は超高圧水銀灯等よりなる蛍光測光用光源、8はコ
レクターレンズ、309はシャッター、10は偏光子、
11は励起フィルター、12は励起光を反射し蛍光を透
過するような特性を有するダイクロイツクミラダで対物
レンズ6を含めこれらで落射蛍光偏光測光用照明系を構
成し、偏光した励起光にて対物レンズ6によ35り試料
5を照明する。又13は励起光を吸収する吸収フィルタ
ー、14は半透過プリズム、15は接眼レンズ、16は
ウオラストンプリズム等の検光子、17a,17bは受
光素子、18a,18bは増巾器、19は演算処理回路
、20は表示装置である。このような光学系において、
前述のようなガンか否かを判定すべき試料をおきまずシ
ヤツタ一3を開き、シヤツタ一9を閉じた状態で、試料
を観察用透過照明系にて照明し、接眼レンズ15にて観
察しながら対物レンズのピット合わせを行ない又、試料
中の測光すべき部分を定める等する。
Further, 7 is a light source for fluorescence photometry such as an ultra-high pressure mercury lamp, 8 is a collector lens, 309 is a shutter, 10 is a polarizer,
11 is an excitation filter, and 12 is a dichroic mirada having the characteristic of reflecting excitation light and transmitting fluorescence, which together with objective lens 6 constitute an illumination system for epifluorescence polarization photometry, and uses polarized excitation light to The sample 5 is illuminated by the objective lens 6 . 13 is an absorption filter that absorbs excitation light; 14 is a semi-transparent prism; 15 is an eyepiece; 16 is an analyzer such as a Wollaston prism; 17a and 17b are light receiving elements; 18a and 18b are amplifiers; The arithmetic processing circuit 20 is a display device. In such an optical system,
First, the sample to be determined as to whether it is cancer or not is placed, the shutter 13 is opened, the shutter 19 is closed, the sample is illuminated with the observation transmission illumination system, and the sample is observed with the eyepiece 15. While doing so, the pits of the objective lens are aligned, and the part of the sample to be photometered is determined.

次にシヤツタ一3を閉じシヤツタ一9を開いて落射蛍光
偏光測光照明系による照明を行なう。これによつて偏光
された励起光にて照明された試料は蛍光を発し、この蛍
光はウオラストンプリズム16を介して受光素子17a
,17bによりその直交する二つの偏光成分が受光され
、増巾器、演算処理回路を通して前述の偏光度Pが算出
され、ガンであるか否かが判定され、その結果が表示装
置20に表示される。以上説明したように本発明のガン
診断装置によれば、その結果力咄動的に表示されること
は勿論顕微鏡による測光を行なうので、試料は極めて微
量で良く、ガンの集団検診に用いれば極めて有効である
Next, the shutter 13 is closed, the shutter 19 is opened, and illumination is performed using an epifluorescence polarization photometric illumination system. The sample illuminated with the polarized excitation light emits fluorescence, and this fluorescence is transmitted through the Wollaston prism 16 to the light receiving element 17a.
, 17b, the two orthogonal polarized components are received, and the aforementioned degree of polarization P is calculated through an amplifier and an arithmetic processing circuit, it is determined whether or not it is cancer, and the result is displayed on the display device 20. Ru. As explained above, according to the cancer diagnostic device of the present invention, the results are not only displayed dynamically but also photometrically measured using a microscope, so the amount of sample required is extremely small, and if used for mass cancer screening, it will be extremely It is valid.

又観察用照明系および蛍光偏光測光用照明系に夫々シヤ
ツタ一を設けて、簡単な操作で両照明系を切換え使用す
るようにしたので、例えば励起光による試料の照明は測
光時のみ行なうことが出来蛍光消光による影響を防止し
得る等、精度の高い判定が可能である。周、実施例の説
明では測光照明系として落射照明を用いているが、透過
照明にすることも可能である。
In addition, a shutter is provided for each of the observation illumination system and the fluorescence polarization photometry illumination system, so that both illumination systems can be switched and used with a simple operation, making it possible, for example, to illuminate the sample with excitation light only during photometry. It is possible to make highly accurate judgments, such as by being able to prevent the effects of fluorescence quenching. Although epi-illumination is used as the photometric illumination system in the description of the embodiment, it is also possible to use transmitted illumination.

又検光子にはウオラストンプリズムを用い二成分に分割
して両成分を検出しているが、検光子を90分回転させ
これと受光素子とを同期させることにより両成分を検出
するようにしても良い。又、蛍光が弱いために受光素子
としては高感度のものが用いられるが、この受光素子に
強い観察用の照明光が入射することは好ましくない。し
たがつて受光素子の前方適宜位置にもシヤツタ一を設け
、測光の時だけ光が受光素子に入射するようにしても良
い。
Also, a Wollaston prism is used in the analyzer to separate the two components and detect both components, but by rotating the analyzer for 90 minutes and synchronizing this with the light receiving element, both components can be detected. It's okay. Further, since the fluorescence is weak, a highly sensitive light receiving element is used, but it is not preferable that strong illumination light for observation be incident on this light receiving element. Therefore, a shutter may be provided at an appropriate position in front of the light receiving element so that light enters the light receiving element only during photometry.

【図面の簡単な説明】[Brief explanation of the drawing]

図面は本発明のガン診断装置の構成を示す図である。 1・・・光源、3・・・シャツタ一、5・・・試料、6
・・・対物レンズ、7・・・測光用光源、9・・・シヤ
ツタ一10・・・偏光子、11・・・励起フイルタ一、
12・・・ダイクロイツクミラー 16・・・検光子、
17a,17b・・・受光素子。
The drawing is a diagram showing the configuration of the cancer diagnostic device of the present invention. 1...Light source, 3...Shaft, 5...Sample, 6
...Objective lens, 7...Photometry light source, 9...Shutter 10...Polarizer, 11...Excitation filter 1,
12...Dichroic mirror 16...Analyzer,
17a, 17b... Light receiving elements.

Claims (1)

【特許請求の範囲】[Claims] 1 シャッターを有する観察用照明系と、シャッター、
偏光子および励起フィルターを有する蛍光偏光測光照明
系と、被検者のリンパ球にガン細胞のタンパクを混ぜた
試料に上記測光照明系により励起光を照射しこれにより
発した蛍光で検光子を通つた光を受光する受光素子と、
上記受光素子よりの電気信号をもとに偏光度を算出する
演算回路とを備えた蛍光偏光測光顕微鏡によるガン診断
装置。
1. An observation illumination system having a shutter;
A fluorescence polarization photometry illumination system with a polarizer and an excitation filter is used to irradiate excitation light onto a sample of lymphocytes from the subject mixed with cancer cell proteins, and the fluorescence emitted by this is passed through an analyzer. a light-receiving element that receives the ivy light;
A cancer diagnostic device using a fluorescence polarization photometric microscope, comprising an arithmetic circuit that calculates the degree of polarization based on the electric signal from the light receiving element.
JP52036200A 1977-04-01 1977-04-01 Cancer diagnostic equipment using a fluorescent polarization photometric microscope Expired JPS5913697B2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP52036200A JPS5913697B2 (en) 1977-04-01 1977-04-01 Cancer diagnostic equipment using a fluorescent polarization photometric microscope
DE19782813317 DE2813317A1 (en) 1977-04-01 1978-03-28 Cancer diagnosis device with direct indication - uses photometric polarisation microscope and fluorescent light reception and analysis system giving stabilised measurements

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP52036200A JPS5913697B2 (en) 1977-04-01 1977-04-01 Cancer diagnostic equipment using a fluorescent polarization photometric microscope

Related Child Applications (1)

Application Number Title Priority Date Filing Date
JP69885A Division JPS60166845A (en) 1985-01-07 1985-01-07 Diagnosis of cancer

Publications (2)

Publication Number Publication Date
JPS53122489A JPS53122489A (en) 1978-10-25
JPS5913697B2 true JPS5913697B2 (en) 1984-03-31

Family

ID=12463082

Family Applications (1)

Application Number Title Priority Date Filing Date
JP52036200A Expired JPS5913697B2 (en) 1977-04-01 1977-04-01 Cancer diagnostic equipment using a fluorescent polarization photometric microscope

Country Status (2)

Country Link
JP (1) JPS5913697B2 (en)
DE (1) DE2813317A1 (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4592361A (en) * 1982-06-28 1986-06-03 The Johns Hopkins University Electro-optical device and method for monitoring instantaneous singlet oxygen concentration produced during photoradiation using pulsed excitation and time domain signal processing
AU562556B2 (en) * 1982-06-28 1987-06-11 Johns Hopkins University, The Electro-optical device for monitoring instantaneous singlet oxygen concentration produced during the treatment of cancer by means of photochemotherapy
JPS60420A (en) * 1983-06-17 1985-01-05 Olympus Optical Co Ltd Fluorescence microphotometric device
AT387860B (en) * 1985-09-16 1989-03-28 Optical Sensing Technology METHOD AND DEVICE FOR TUMOR DIAGNOSIS BY MEANS OF SERA
EP1610671B1 (en) 2003-03-18 2013-08-21 The General Hospital Corporation Polarized light devices and methods
CA2539184A1 (en) 2003-09-19 2005-03-31 The General Hospital Corporation Fluorescence polarization imaging devices and methods
DE102011018725A1 (en) * 2011-04-20 2012-10-25 Carl Zeiss Jena Gmbh Optical arrangement with luminometer for detecting luminescence of samples, has emission filter formed as absorbent color filter without reflective layer, particularly without dichroic layers, where emission filter is arranged in beam path

Also Published As

Publication number Publication date
DE2813317A1 (en) 1978-10-05
JPS53122489A (en) 1978-10-25

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