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CN109540890A - A kind of DNA quantitative analysis method based on microcytoscope image - Google Patents

A kind of DNA quantitative analysis method based on microcytoscope image Download PDF

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Publication number
CN109540890A
CN109540890A CN201811425981.9A CN201811425981A CN109540890A CN 109540890 A CN109540890 A CN 109540890A CN 201811425981 A CN201811425981 A CN 201811425981A CN 109540890 A CN109540890 A CN 109540890A
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China
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cell
image
quantitative analysis
microcytoscope
analysis method
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CN201811425981.9A
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Chinese (zh)
Inventor
贺权源
刘朝前
李仕能
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Hunan Pisen Biotechnology Co Ltd
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Hunan Pisen Biotechnology Co Ltd
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Priority to CN201811425981.9A priority Critical patent/CN109540890A/en
Publication of CN109540890A publication Critical patent/CN109540890A/en
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    • 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
    • G01N21/84Systems specially adapted for particular applications

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

A kind of DNA quantitative analysis method based on microcytoscope image of the present invention, it will be directly from the information RAW file obtained on CCD or CMOS as the picture in DNA quantitative analysis, it needs to increase compensation calculation to avoid in calculating IOD value, and it is made mistakes, to greatly improve the accuracy that DNA is quantitatively calculated, and when increasing or decreasing incident intensity, keep its result more stable.

Description

A kind of DNA quantitative analysis method based on microcytoscope image
Technical field
The present invention relates to technical field of image processing more particularly to a kind of DNA based on microcytoscope image quantitatively to divide Analysis method.
Background technique
Cytolgical examination is the main path of the exemplary cancers early screening such as current cervical carcinoma, carcinoma of mouth, is used in early days Doctor blade technique is because cheap and universal, but its rate of missed diagnosis is high, although the liquid-based tabletting technology being widely used since 21 century changes It has been apt to production effect, but has been provided after needing veteran pathologist directly to observe under the microscope the form of cell Diagnostic result, low efficiency, and be easy to receive the influence of subjectivity, repeatability is poor.DNA ploidy body analytical technology is then automatically to thin Nucleus in born of the same parents' image is analyzed, and then quantitative measurment goes out chromosome or DNA content in nucleus, and provides judgement automatically As a result.Compared with conventional cell method, which has the characteristics that sensibility is high, high-efficient and favorable repeatability, Ke Yiyou Effect solves base and lacks experienced this contradiction of pathologist, is with a wide range of applications.
It is DNA ploidy body analytical technology that fast and accurately diploid cell core in cell image, which is split and is identified, It is crucial.Existing method and system are extracted in segmentation using simple images dividing methods such as Threshold segmentation, morphological methods Candidate region, various shape, Texture eigenvalue and the trained classifier for then extracting candidate region complete oneself of nucleus type Dynamic identification.However such methods are when handling more intensive and nucleus the intensity profile variation of nucleus distribution greatly A large amount of diploid cell can be missed, and often extracts all features in each candidate region in identification, in candidate region number Time-consuming in the case that amount is big.
Steps are as follows in the source of picture in currently used DNA quantitative analysis method: incident light → glass slide → transmitted light → CCD (or CMOS), last CCD (or CMOS) carry out color rendition to picture, form a color image or black picture, When calculating the IOD value in DNA, need to picture carry out color rendition during to the rgb value of picture do compensation or It is compensated when calculating the gray value of picture after gray processing, therefore is unable to get accurately IOD value, to cause to will affect in DNA Accuracy when quantitative analysis.
Summary of the invention
The purpose of the present invention is the picture using the information that is directly obtained from CCD or CMOS as DNA quantitative analysis, with Avoid a kind of DNA quantitative analysis method based on microcytoscope image that error is generated because calculating need to be compensated.
Technical solution of the invention is as follows:
A kind of DNA quantitative analysis method based on microcytoscope image, comprises the following steps that
Step 1: the slide for being loaded with cell being positioned to the detection center of microscope carrier, so that cell-rich areas position Under microscopical observation sight;
Step 2: cell plane being focused and taken pictures, and the image after taking pictures is saved;
Step 3: analyze the image after taking pictures: acquisition normal diploid, abnormal times of body, lymphocyte, other are thin Born of the same parents' image carries out the integral optical density value of calculating nucleus after image segmentation to it.
Step 4: taxonomic revision is carried out to the cell after calculating, calculate all cells parameter (comprising IOD, area, etc. Multiple parameters), the IOD value of all cells is ranked up from big to small, abnormal times of body is selected after sequence, and (IOD value is greater than two times Body IOD 2.5, area be greater than diploid) cell.
As further scheme of the invention: in the step 2, when taking pictures to cell plane, using with 3 times of essences The mode at true focus steps interval is taken pictures;
Further, in the step 2, after taking pictures to cell liquid, with the information directly obtained from CCD or CMOS RAW file is picture.
As further scheme of the invention: in the step 4, the calculating of the IOD value only needs to calculate every in nucleus The sum of transmitted intensity/incident intensity logarithm of a point.
The beneficial effects of the present invention are: 1, RAW file is almost untreated and directly obtains from CCD or CMOS Information;2, there is no white balance settings for RAW file, but true data are not also changed, and can arbitrarily adjust colour temperature And white balance, do not have image quality loss;3, by RAW file as the benefit of DNA quantitative analysis picture it is clear that transmission Luminous intensity and incident intensity all do not compensate, and greatly improve the accuracy that DNA is quantitatively calculated, and increase or decrease into When penetrating luminous intensity, keep its result more stable.
Detailed description of the invention
To describe the technical solutions in the embodiments of the present invention more clearly, make required in being described below to embodiment Attached drawing is briefly described, it should be apparent that, drawings in the following description are only some embodiments of the invention, for For those of ordinary skill in the art, without creative efforts, it can also be obtained according to these attached drawings other Attached drawing, in which:
Fig. 1 is method flow schematic diagram of the invention.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, to the detailed description of the invention.
As shown in Figure 1, a kind of DNA quantitative analysis method based on microcytoscope image disclosed by the invention, including step It is rapid as follows:
Step 1: the slide for being loaded with cell being positioned to the detection center of microscope carrier, so that cell-rich areas position Under microscopical observation sight;
Step 2: cell plane being focused and taken pictures, and the image after taking pictures is saved, and from CCD or CMOS The message file directly obtained is directly as analysis picture;
Further, the message file directly obtained from CCD or CMOS is RAW file.
Step 3: analyze the image after taking pictures: acquisition normal diploid, abnormal times of body, lymphocyte, other are thin Born of the same parents' image carries out the integral optical density value of calculating nucleus after image segmentation to it;
Step 4: taxonomic revision is carried out to the cell after calculating, calculate all cells parameter (comprising IOD, area, etc. Multiple parameters);
Further, calculated all cellular IOD values are less than maximum to select from being ranked up to small sequence greatly Diploid cell IOD value and the cell for being greater than minimum diploid cell IOD value;
As further embodiment of the present invention: the IOD value sequence and calculating DNA index comprise the concrete steps that: 1, by institute There are cellular IOD values to be ranked up from as low as big sequence, 2, select and be less than maximum diploid cell IOD value and be greater than minimum two The cell of times body cell IOD value, 3, calculate the cell quantity of per unit IOD, 4, using per unit cell quantity as ordinate, with IOD value is horizontal seatpost, does cell scatter plot, 5, with gaussian curve approximation, 6, calculated curve μ value, μ value is then diploid cell Normalize IOD value.
As further embodiment of the present invention: normalizing IOD when the IOD of cell is more than or equal to 2.5 times of diploid cells When value, which is identified as suspected lesion cell automatically.
The above content is merely illustrative of the technical solution of the present invention, rather than limiting the scope of the invention, this field Those of ordinary skill to technical solution of the present invention carry out it is simple modification or equivalent replacement, all without departing from the technology of the present invention The spirit and scope of scheme.

Claims (5)

1. a kind of DNA quantitative analysis method based on microcytoscope image, comprises the following steps that
Step 1: the slide for being loaded with cell being positioned to the detection center of microscope carrier, is shown so that cell-rich areas is located at Under the observation sight of micro mirror;
Step 2: cell plane being focused and taken pictures, and the image after taking pictures is saved;
Step 3: the image after taking pictures being analyzed: acquisition normal diploid, abnormal times of body, lymphocyte, other cytological maps Picture carries out the integral optical density value of calculating nucleus after image segmentation to it;
Step 4: taxonomic revision is carried out to the cell after calculating, calculates the parameter of all cells, to the IOD values of all cells from Arrive greatly it is small be ranked up, the cell of abnormal times of body is selected after sequence.
2. a kind of DNA quantitative analysis method based on microcytoscope image according to claim 1, it is characterised in that: In the step 2, when taking pictures to cell plane, uses and taken pictures in a manner of 3 times of vernier focusing step intervals.
3. a kind of DNA quantitative analysis method based on microcytoscope image according to claim 2, it is characterised in that: In the step 2, after taking pictures to cell plane, with the message file that is directly obtained from CCD or CMOS directly as point Analyse picture.
4. a kind of DNA quantitative analysis method based on microcytoscope image according to claim 3, it is characterised in that: The message file directly obtained from CCD or CMOS is RAW file.
5. a kind of DNA quantitative analysis method based on microcytoscope image according to claim 1, it is characterised in that: In the step 4, the calculating of the IOD value only needs to calculate transmitted intensity/incident intensity pair of each point in nucleus The sum of numerical value.
CN201811425981.9A 2018-11-27 2018-11-27 A kind of DNA quantitative analysis method based on microcytoscope image Withdrawn CN109540890A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109182453A (en) * 2018-09-13 2019-01-11 湖南品胜生物技术有限公司 A kind of control method of DNA ploidy body analytical equipment
CN112378727A (en) * 2020-12-14 2021-02-19 湖南莱博赛医用机器人有限公司 DNA ploid quantitative analysis device based on machine vision and application method thereof
CN112651368A (en) * 2020-12-30 2021-04-13 宁波江丰生物信息技术有限公司 DNA ploidy analysis method for large-size cell microscopic image

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109182453A (en) * 2018-09-13 2019-01-11 湖南品胜生物技术有限公司 A kind of control method of DNA ploidy body analytical equipment
CN112378727A (en) * 2020-12-14 2021-02-19 湖南莱博赛医用机器人有限公司 DNA ploid quantitative analysis device based on machine vision and application method thereof
CN112651368A (en) * 2020-12-30 2021-04-13 宁波江丰生物信息技术有限公司 DNA ploidy analysis method for large-size cell microscopic image

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Application publication date: 20190329