WO2012126346A1 - Procédé pour la détermination rapide de la teneur en adn dans un noyau cellulaire - Google Patents

Procédé pour la détermination rapide de la teneur en adn dans un noyau cellulaire Download PDF

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Publication number
WO2012126346A1
WO2012126346A1 PCT/CN2012/072540 CN2012072540W WO2012126346A1 WO 2012126346 A1 WO2012126346 A1 WO 2012126346A1 CN 2012072540 W CN2012072540 W CN 2012072540W WO 2012126346 A1 WO2012126346 A1 WO 2012126346A1
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WO
WIPO (PCT)
Prior art keywords
cells
dna content
image
nucleus
cell
Prior art date
Application number
PCT/CN2012/072540
Other languages
English (en)
Chinese (zh)
Inventor
孙小蓉
黄么姑
Original Assignee
武汉兰丁医学高科技有限公司
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 武汉兰丁医学高科技有限公司 filed Critical 武汉兰丁医学高科技有限公司
Publication of WO2012126346A1 publication Critical patent/WO2012126346A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites

Definitions

  • the invention belongs to the field of nucleotide detection, and in particular relates to a method for rapidly determining DNA content in a nucleus. Background technique
  • the measurement of DNA in the nucleus of a living body is mostly measured by an optical method.
  • the general method is to combine the dye or developer with the DNA specificity in the nucleus and measure the optical density in the nucleus under illumination.
  • the DNA content in the nucleus is increased, the DNA binding dye or color developer is increased, and the optical density is increased.
  • the amount of DNA in the nucleus can be determined by image analysis to determine whether the cell is cancerous.
  • the clinician examines the cells under the microscope, in addition to observing the nuclei, the color and size of the cytoplasm are also observed.
  • the doctor determines whether the cell is cancerous and the type and classification of the cell.
  • the chromosome is deeply stained, the arrangement is disordered, and when the ratio of the nucleus to the cytoplasm is reduced, it is possible to discriminate that the cell is cancerous.
  • the present invention aims to provide a method for rapidly determining the DNA content in the nucleus by improving the conventional DNA content determination technique in the human nucleus.
  • the object of the present invention is achieved as follows: A method for rapidly measuring the DNA content in a nucleus, characterized in that: the nucleus is first stained with Feulgen, and then the cytoplasm is stained with EA50 or eosin, after two staining, the glass is The on-chip samples are scanned using a computer image analysis system, and both grayscale and color images are acquired and registered; The morphological characteristics of the specification, classify the cells under the field of view, select normal lymphocytes as standard cells for calculating DNA content; select epithelial cells in each field of view, and calculate each of the slides according to the DNA content of the standard cells. The DNA content of the epithelial cells, record their location, and save the cell image. Gray-scale images and color images are provided for the same area of view for computer image analysis system analysis, and color images are provided for morphological observation and doctor review.
  • the grayscale image described above is a picture of the nuclei taken by a black and white camera at a wavelength of 590 light.
  • a method for simultaneously acquiring a grayscale image and a color image wherein: the black and white and the color double microscope camera are used to simultaneously control the image acquisition.
  • An automatic platform control method for automatically locating cells characterized in that: according to the coordinates of the cells on the slide, the field of view of the cells is positioned, and the platform is moved to the field of view for microscopic recombination.
  • a method for establishing a clinically suspected DNA content case database characterized in that: clinically confirmed images of suspected cells (including nuclei and cytoplasm) are stored and analyzed for comparison with the cells to be tested.
  • a method for determining whether a DNA content of a cell is stable or not is characterized in that: the DNA content of the lymphocyte is relatively stable, and the DNA content of the lymphocyte is used as a criterion for judging.
  • the method for determining the DNA content in the nucleus of the present invention has the following advantages and significant progress: it can quickly determine the DNA content in the nucleus, and can facilitate the clinician to observe the cell morphology (nucleus and cytoplasm), thereby accurately diagnosed at an early stage. The patient's condition. DRAWINGS
  • Figure 1 A grayscale image taken with a black-and-white camera under a 20x microscope.
  • the framed portion is the cell that is detected by the system to be cancerous.
  • Figure 2 A color image taken with a color camera under a 20x microscope.
  • the part that is framed is the cell that is detected by the system to be cancerous.
  • the present invention first classifies cells in the field of view according to the shape characteristics of the cells, and divides the cells into three types: epithelial cells, lymphocytes, and other cells.
  • the DNA content in the lymphocyte nucleus is relatively stable, so when determining whether a living body cell is cancerous, the content of DNA in the lymphocyte nucleus can be used as a reference. This hair The book explains the content of nuclear DNA in epithelial cells through image analysis system.
  • the ratio of the content of nuclear DNA in epithelial cells to the content of lymphocyte nuclear DNA reaches a certain threshold, it indicates that the cells may be cancerous.
  • the system can automatically scan the entire slide, use color and black-and-white camera to take pictures at the same time, analyze the image to count the number of cells that may be cancerous and locate the field of view of the cells. The doctor can easily find the cells that may be cancerous. The field of view, the color image is analyzed and reviewed.
  • the system has the following improvements:
  • the black and white camera takes pictures of the nuclei under a certain wavelength (590 ⁇ ) illumination to determine the DNA content, and the color camera is used for the nuclei and Observation of cytoplasm morphology.
  • Suspected cell morphology analysis The clinically confirmed images of suspected cells (including nuclei and cytoplasm) are stored and analyzed for characteristic values such as nucleoplasmic ratio, nuclear morphology and color.
  • the cells in the field of view are classified, a certain number of fields of view are scanned, and better lymphocytes are selected as standard cells for calculating DNA content;
  • a histogram of the DNA content of epithelial cells is counted.
  • the nucleus and cytoplasm of each cell to be tested are displayed in color images, and the corresponding visual field map is saved.
  • the morphology of the suspicious cells stored is compared, and the abnormality of the cells to be tested is determined. See Figure 1 2;

Abstract

La présente invention concerne un procédé de détermination rapide de la teneur en ADN, consistant premièrement à appliquer un marquage de Feulgen aux noyaux cellulaires, puis marquer le cytoplasme avec EA50 ou de l'éosine. Après que le marquage a eu lieu deux fois, le procédé consiste à scanner l'échantillon sur une lame à l'aide d'un système d'analyse d'image par informatique et acquérir et aligner une image en niveaux de gris et une image en couleur ; trier les cellules dans le champ de vision selon les caractéristiques morphologiques des cellules, et sélectionner les lymphocytes normaux en tant que cellules standard pour le calcul de la teneur en ADN. Le procédé consiste également à sélectionner des cellules épithéliales dans chaque champ de vision, calculer la teneur en ADN dans chaque épithélium sur la lame selon la teneur en ADN des cellules standards, enregistrer la position de chaque épithélium et stocker l'image des cellules.
PCT/CN2012/072540 2011-03-22 2012-03-19 Procédé pour la détermination rapide de la teneur en adn dans un noyau cellulaire WO2012126346A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201110068228.0 2011-03-22
CN 201110068228 CN102181534A (zh) 2011-03-22 2011-03-22 一种快速测定细胞核内dna含量的方法

Publications (1)

Publication Number Publication Date
WO2012126346A1 true WO2012126346A1 (fr) 2012-09-27

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2012/072540 WO2012126346A1 (fr) 2011-03-22 2012-03-19 Procédé pour la détermination rapide de la teneur en adn dans un noyau cellulaire

Country Status (2)

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CN (1) CN102181534A (fr)
WO (1) WO2012126346A1 (fr)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102181534A (zh) * 2011-03-22 2011-09-14 武汉兰丁医学高科技有限公司 一种快速测定细胞核内dna含量的方法
CN108956398B (zh) * 2018-07-04 2020-10-16 东南大学 基于图像分析定量细胞内铁纳米颗粒含量的方法
CN112378727A (zh) * 2020-12-14 2021-02-19 湖南莱博赛医用机器人有限公司 基于机器视觉的dna倍体定量分析装置及其应用方法
CN112750493B (zh) * 2020-12-22 2024-05-03 深思考人工智能机器人科技（北京）有限公司 基于巴氏染色方式的dna倍体定量分析方法及系统
CN113178228B (zh) * 2021-05-25 2023-02-10 郑州中普医疗器械有限公司 基于细胞核dna分析的细胞分析方法、计算机设备、存储介质

Citations (3)

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Publication number Priority date Publication date Assignee Title
CN1185304A (zh) * 1996-12-17 1998-06-24 顾康庭 病理工作站系统的组成方法
CN101560544A (zh) * 2008-04-18 2009-10-21 麦克奥迪实业集团有限公司 一种细胞综合检测方法
CN102181534A (zh) * 2011-03-22 2011-09-14 武汉兰丁医学高科技有限公司 一种快速测定细胞核内dna含量的方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1185304A (zh) * 1996-12-17 1998-06-24 顾康庭 病理工作站系统的组成方法
CN101560544A (zh) * 2008-04-18 2009-10-21 麦克奥迪实业集团有限公司 一种细胞综合检测方法
CN102181534A (zh) * 2011-03-22 2011-09-14 武汉兰丁医学高科技有限公司 一种快速测定细胞核内dna含量的方法

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ZHANG, WEI ET AL.: "Screening for Early Cervical Cancer and Precancerous Cervical Lesions by DNAPloid Analysis", CHINA CANCER, vol. 16, no. 7, 31 July 2007 (2007-07-31), pages 508 - 511 *

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