WO2012126346A1 - Method for rapidly determining dna content in cell nucleus - Google Patents

Method for rapidly determining dna content in cell nucleus 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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cells
dna content
image
nucleus
cell
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PCT/CN2012/072540
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French (fr)
Chinese (zh)
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孙小蓉
黄么姑
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武汉兰丁医学高科技有限公司
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    • 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

The present invention provides a method for rapidly determining DNA content, comprising first applying Feulgen staining to the cell nuclei, then staining the cytoplasm with EA50 or eosin, after staining two times, scanning the sample on a slide with a computer image analysis system, and acquiring and aligning a grayscale image and a color image; sorting the cells in the field of vision according to the morphological features of the cells, and selecting normal lymphocytes as standard cells for calculating the DNA content; selecting epithelial cells in each field of vision, calculating the DNA content in each epithelium on the slide according to the DNA content of the standard cells, recording the position of each epithelium, and storing the cell picture.

Description

说 明 书  Description
一种快速测定细胞核内 DNA含量的方法  Method for rapidly measuring DNA content in nuclear cells
技术领域 Technical field
本发明属于核苷酸检测领域， 尤其涉及一种快速测定细胞核内 DNA含量的方法。 背景技术  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
当生物体细胞发生癌变时， 细胞核内的 DNA含量会增加。 因此， 可以根据细胞核内 DNA含量是否显著增加来判断细胞是否发生癌变。 生物体细胞核内 DNA的测定大多是通过 光学方法来测定的。一般方法是将染料或显色剂与细胞核内 DNA特异性相结合， 在光照下 测定细胞核内光密度。 当细胞核内 DNA含量增高时， DNA结合染料或显色剂增多， 其光学 密度增高。 利用这个原理， 可以通过图像分析测定细胞核内 DNA的含量， 来判断细胞是否 发生癌变。 另外， 临床医生在显微镜下检査细胞时， 除对细胞核进行观察外， 还要观察细 胞浆的颜色及大小。 医生通过观测细胞核内染色体粗细、 染色体排列以及细胞核与细胞浆 的比值， 来判断细胞是否癌变以及细胞的分型和分类。 当染色体深染， 排列紊乱并且当细 胞核与细胞浆的比例縮小时， 就可判别出细胞出现了癌变的可能。  When a living body cells become cancerous, the DNA content in the nucleus increases. Therefore, whether or not the cells are cancerous can be judged based on whether or not the DNA content in the nucleus is significantly increased. 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. When the DNA content in the nucleus is increased, the DNA binding dye or color developer is increased, and the optical density is increased. Using this principle, the amount of DNA in the nucleus can be determined by image analysis to determine whether the cell is cancerous. In addition, when the clinician examines the cells under the microscope, in addition to observing the nuclei, the color and size of the cytoplasm are also observed. By observing the chromosome thickness, chromosome arrangement, and ratio of nucleus to cytoplasm in the nucleus, the doctor determines whether the cell is cancerous and the type and classification of the cell. When 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.
目前， 医学上用显微镜确诊的肿瘤患者多为晚期肿瘤病人， 然而对于早期的肿瘤患者 却很难发现， 原因在于早期患者的肿瘤细胞核内 DNA含量增加不显著， 因此很多早期的肿 瘤患者错过了最佳治疗时期。 另外， 医生的经验也决定了医生的诊断水平。 正因为如此， 使得不同的医生在诊断上存在着一定差别。 同时， 每个样本有较多的细胞， 在显微镜下要 观察大量的视野才能看完一个玻片， 在样本量较大时容易产生视觉疲劳。  At present, medically diagnosed tumor patients are mostly advanced tumor patients, but it is difficult to find for early tumor patients. The reason is that the DNA content in the tumor cells of early patients is not significantly increased, so many early tumor patients missed the most. Good treatment period. In addition, the doctor's experience also determines the doctor's diagnostic level. Because of this, there are some differences in the diagnosis of different doctors. At the same time, each sample has more cells. Under the microscope, a large amount of field of view is required to view a slide, and visual fatigue is likely to occur when the sample size is large.
如何做到将细胞样本处理后， 既能测定细胞核内 DNA含量， 又能方便临床医生观察细 胞形态 （细胞核及细胞浆）， 从而在早期准确地诊断出患者的病情， 这是一个长期未解决 的难题。 发明内容  How to treat the cell sample can not only determine the DNA content in the nucleus, but also facilitate the clinician to observe the cell morphology (nucleus and cytoplasm), so that the patient's condition can be accurately diagnosed at an early stage. This is a long-term unsolved problem. problem. Summary of the invention
鉴于现有技术的不足，本发明的目的在于通过对传统的人体细胞核内 DNA含量测定技 术进行改进， 提供了一种快速测定细胞核内 DNA含量的方法。  In view of the deficiencies of the prior art, 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.
本发明的目的是这样实现的： 一种快速测定细胞核内 DNA含量的方法， 其特征在于： 先对细胞核用 Feulgen染色， 然后再用 EA50或伊红染色细胞浆， 经过两次染色后， 将玻 片上的样本用计算机图像分析系统进行扫描， 同时获取灰度与彩色图像并配准； 根据细胞 说 明 书 的形态特征， 将视野下的细胞进行分类， 从中选取正常的淋巴细胞作为计算 DNA含量的标 准细胞； 选出每个视野中的上皮细胞， 根据标准细胞的 DNA含量计算出玻片上的每个上皮 细胞的 DNA含量， 记录其所在位置， 并将细胞图片保存。对同一区域视野提供灰度图像与 彩色图像供计算机图像分析系统分析， 并提供彩色图像供形态学观测和医生复查。 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.
以上所述的灰度图像是通过黑白摄像头在波长 590 光照下的细胞核取图。  The grayscale image described above is a picture of the nuclei taken by a black and white camera at a wavelength of 590 light.
另外， 本发明还提供了如下技术方案：  In addition, the present invention also provides the following technical solutions:
一种同时获取灰度图像与彩色图像的方法， 其特征在于： 釆用黑白与彩色双显微镜摄 像头进行同时控制取图。  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.
一种自动定位细胞的自动平台控制方法， 其特征在于： 根据细胞在玻片上的坐标， 定 位出细胞所在的视野， 并使平台移动到该视野进行镜下复合。  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.
一种建立临床可疑 DNA含量病例数据库的方法， 其特征在于： 将临床上证实的可疑 细胞图像 （包括细胞核和细胞浆） 进行储存及分析其特征， 用于与被测细胞对比。  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.
一种细胞 DNA含量稳定与否的判定方法， 其特征在于： 根据淋巴细胞核 DNA含量相 对稳定， 以淋巴细胞核 DNA含量作为判断的标准。  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.
本发明涉及的测定细胞核内 DNA含量的方法具有如下优点和显著的进步：既能快速测 定细胞核内 DNA含量， 又能方便临床医生观察细胞形态 （细胞核及细胞浆）， 从而在早期 准确地诊断出患者的病情。 附图说明  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
图 1 用黑白摄像头在 20倍显微镜下拍摄的灰度图像， 被框出的部分是被系统检测有 癌变可能的细胞。  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.
图 2 用彩色摄像头在 20倍显微镜下拍摄的彩色图像， 被框出的部分是被系统检测有 癌变可能的细胞。  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.
图 3 实施例 1的方法经过图像分析系统得出的报告。 具体实施方式  Figure 3. Report of the method of Example 1 through an image analysis system. detailed description
由于生物体内， 上皮细胞的面积比淋巴细胞大， 同时淋巴细胞的形状较圆， 而上皮细 胞的形状不确定。 所以， 本发明首先根据细胞的形状特征， 对视野下的细胞进行分类， 将 细胞分为三类： 上皮细胞、 淋巴细胞和其它细胞。 而淋巴细胞核中的 DNA含量相对稳定， 所以在测定生物体细胞是否发生癌变时， 可以以淋巴细胞核中 DNA的含量作为基准。本发 说 明 书 明通过图像分析系统计算上皮细胞核 DNA的含量，若上皮细胞核 DNA的含量与淋巴细胞核 DNA的含量的比值达到一定的阈值， 则说明此细胞有癌变的可能。 本系统可以自动的扫描 整个玻片， 采用彩色与黑白摄像头进行同时取图， 通过分析图像统计出可能发生癌变的细 胞数量并定位出细胞所在的视野， 医生可以很方便地找到可能发生癌变的细胞所在的视 野， 对彩色图像进行分析复查。 Due to the fact that the area of epithelial cells is larger than that of lymphocytes in the living body, the shape of lymphocytes is relatively round, and the shape of epithelial cells is uncertain. Therefore, 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. If 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.
不同于巳有 DNA系统， 本系统有如下几方面的改进：  Unlike the DNA system, the system has the following improvements:
显微镜摄像头的改进： 在一台显微镜下， 同时用彩色摄像头和黑白摄像头各一个， 黑 白摄像头对一定波长 （590ηπ ) 光照下的细胞核取图， 从而进行 DNA含量的测定， 彩色摄 像头用于对细胞核及细胞浆形态的观察。  Improvement of the microscope camera: Under one microscope, one color camera and one black and white camera are used at the same time. The black and white camera takes pictures of the nuclei under a certain wavelength (590ηπ) illumination to determine the DNA content, and the color camera is used for the nuclei and Observation of cytoplasm morphology.
染色方法的改进： 先用 Feulgen染色细胞核做 DNA定量测定， 后用 EA50或伊红染色 细胞浆便于形态测定和医生复查， 以往很少人用此方法。  Improvement of staining method: Firstly, the cells were stained with Feulgen for quantitative determination of DNA, and then stained with EA50 or eosin for morphological determination and doctor review. This method has rarely been used in the past.
可疑细胞形态分析： 将临床上证实的可疑细胞图像 （包括细胞核和细胞浆）， 进行储 存及分析其特征值， 如核浆比例、 细胞核形态及颜色等。  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.
软件上的改进： 既可实现 DNA定量测定， 又实现黑白与彩色摄像头同时摄像而显示彩 色图像； 再者， 实现被测细胞与可疑细胞对比。  Software improvements: It can realize the quantitative measurement of DNA, and realize the simultaneous display of black and white and color cameras to display color images; in addition, the measured cells are compared with suspicious cells.
以下是本发明的具体实施例， 对本发明的技术方案做进一歩作描述， 但是本发明的保 护范围并不限于这些实施例。凡是不背离本发明构思的改变或等同替代均包括在本发明的 保护范围之内。  The following is a specific embodiment of the present invention, and the technical solution of the present invention will be further described, but the scope of protection of the present invention is not limited to these embodiments. Any changes or equivalents that do not depart from the inventive concept are included in the scope of the present invention.
实施例 1 细胞核内 DNA含量的测定  Example 1 Determination of DNA content in the nucleus
( 1 ) 用 Feulgen染色方法对细胞核进行染色；  (1) staining the nucleus with Feulgen staining;
(2 ) 用 EA50或伊红对细胞浆进行染色， 染色方法根据样本而定： 如果为鳞状上皮， 用 EA50染色； 如果是其他样本， 用伊红染色；  (2) staining the cytoplasm with EA50 or eosin according to the sample: if it is squamous epithelium, stain with EA50; if it is other samples, stain with eosin;
(3 ) 将染色后的玻片放在显微镜下， 对玻片进行扫描， 同时获取灰度与彩色图像并配 准；  (3) Place the stained slide under the microscope, scan the slide, and obtain the grayscale and color images and register them;
(4) 根据细胞的形态特征， 将视野下的细胞进行分类， 扫描一定数量的视野， 从中选 取较好的淋巴细胞作为计算 DNA含量的标准细胞；  (4) According to the morphological characteristics of the cells, 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;
( 5 ) 计算基准细胞 （淋巴细胞） 中 DNA的含量， 假设为 2c单位；  (5) Calculate the amount of DNA in the reference cells (lymphocytes), assuming 2c units;
(6 ) 选出每个视野中的上皮细胞， 同时计算出每个上皮细胞的 DNA含量， 同时记录每 个细胞所在的位置， 并将细胞图片保存；  (6) Select epithelial cells in each field of view, calculate the DNA content of each epithelial cell, record the location of each cell, and save the cell image;
(7) 重复第 6步， 直至系统扫描完整个玻片； 说 明 书 (7) Repeat step 6 until the system scans the entire slide; Instruction manual
( 8) 统计出上皮细胞 DNA含量的直方图。 同时将每个被测细胞的胞核及胞浆用彩色图 像显示出来， 并保存相应的视野图， 同时对比己储存的可疑细胞形态， 判断出被测细 胞是否有异常， 异常细胞参见图 1-2; (8) A histogram of the DNA content of epithelial cells is counted. At the same time, the nucleus and cytoplasm of each cell to be tested are displayed in color images, and the corresponding visual field map is saved. At the same time, the morphology of the suspicious cells stored is compared, and the abnormality of the cells to be tested is determined. See Figure 1 2;
(9) 医生通过此系统定位出可疑细胞 （DNA含量异常或细胞形态异常） 所在的视野， 直接在相应的显微镜视野下进行复查；  (9) The doctor uses this system to locate the field of view of suspicious cells (abnormal DNA content or abnormal cell morphology), and directly review it under the corresponding microscope field;
( 10) 系统自动打印诊断报告， 参见图 3。  (10) The system automatically prints a diagnostic report, see Figure 3.

Claims

权 利 要 求 书 Claim
1、 一种快速测定细胞核内 DNA含量的方法， 其特征在于： 先对细胞核用 Feulgen染色， 然后再用 EA50或伊红染色细胞浆， 经过两次染色后， 将玻片上的样本用计算机图像分析系统 进行扫描， 同时获取灰度与彩色图像并配准； 所述的配准步骤为： 根据细胞的形态特征， 将 视野下的细胞进行分类， 从中选取正常的淋巴细胞作为计算 DNA含量的标准细胞； 选出每个 视野中的上皮细胞， 根据标准细胞的 DNA含量计算出玻片上的每个上皮细胞的 DNA含量， 记 录其所在位置， 并将细胞图片保存。 A method for rapidly measuring 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 sample on the slide is analyzed by computer image. The system scans and acquires grayscale and color images and registers them. The registration step is as follows: According to the morphological characteristics of the cells, the cells in the field of view are classified, and normal lymphocytes are selected as standard cells for calculating DNA content. The epithelial cells in each field of view were selected, and the DNA content of each epithelial cell on the slide was calculated based on the DNA content of the standard cells, the position was recorded, and the cell image was saved.
2、 如权利要求 1所述的快速测定细胞核内 DNA含量的方法， 其特征在于： 所述的灰度图 像是通过黑白摄像头在波长 590nm光照下的细胞核取图。  2. A method for rapidly determining DNA content in a nucleus according to claim 1, wherein: said grayscale image is a nuclear image taken by a black and white camera at a wavelength of 590 nm.
3、 一种细胞核与细胞浆的染色方法， 其特征在于： 先对细胞核用 Feulgen染色， 然后再 用 EA50或伊红染色细胞浆。  3. A method for staining a nucleus and a cytoplasm, characterized in that: the nucleus is first stained with Feulgen, and then the cytoplasm is stained with EA50 or eosin.
4、 一种快速测定细胞核 DNA含量的方法， 其特征在于： 对两次染色后的样本用计算机 图像分析系统进行扫描， 同时获取灰度与彩色图像并配准， 对同一区域视野提供灰度图像与 彩色图像供计算机图像分析系统分析， 根据细胞的光学密度来计算细胞核的 DNA含量， 并 提供彩色图像供形态学观测和医生复查。  4. A method for rapidly determining the nuclear DNA content of a cell, characterized in that: the sample after two staining is scanned by a computer image analysis system, and the gray scale and the color image are simultaneously acquired and registered, and the gray image is provided for the field of view of the same region. The color image is analyzed by a computer image analysis system, and the DNA content of the nucleus is calculated according to the optical density of the cell, and a color image is provided for morphological observation and doctor review.
5、 一种同时获取灰度图像与彩色图像的方法， 其特征在于： 采用黑白与彩色双显微镜摄 像头进行同时控制取图。  5. 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.
6、 一种自动定位细胞的自动平台控制方法， 其特征在于： 根据细胞在玻片上的坐标， 定 位出细胞所在的视野， 并使平台移动到该视野进行镜下复合。  6. 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.
7、 一种建立临床可疑 DNA含量病例数据库的方法， 其特征在于： 将临床上证实的可疑 细胞图像 （包括细胞核和细胞浆） 进行储存及分析其特征， 用于与被测细胞对比。  7. 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.
8、 一种细胞 DNA含量稳定与否的判定方法， 其特征在于： 根据淋巴细胞核 DNA含量相 对稳定， 以淋巴细胞核 DNA含量作为判断的标准。  8. A method for determining whether a cell DNA content is stable or not, characterized in that: the DNA content of the lymphocyte nuclear is relatively stable, and the DNA content of the lymphocyte nuclear is used as a criterion for judging.
9、 一种细胞分类的方法， 其特征在于： 根据淋巴细胞与上皮细胞的不同形态特征对细胞 进行分类。  9. A method of cell sorting, characterized in that: cells are classified according to different morphological characteristics of lymphocytes and epithelial cells.
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