Methodology And Key Technology For Cell DNA Quantitative Analysis In Microscopic Spectral Imaging

Recently,it is increasingly severe to prevent and treat cervical cancer which the most import means to prevent and control are early-screening and diagnosis.In China,the effictive cervical cancer screening program was based on liquid-based cytology combined with cell DNA quantitative analysis techniques.Howerver,these two methods need to be analyzed on different smears which exits some shortcomings such as waste of resources,low efficiency and high error rate.Therefore,it is urgent to study a method which can perfect the fusion of two technologies to achieve efficient and accurate screening.Based on existion products and combined with market demand,this research studied three key independent and coherent techniques for specimen screening,speciment staining and speciment diagnosis in cytological screening and achieved the following achievements:(1)Proposed a cytological screening method based on composite staining and microspectroscopic imaging in which only one sample is taken to prepare a smear,and the two staining methods act on the same cell,so that the DAN can be quantitatively diagnosed and the cell morphology can also be used for TBS classification.Moreover,in the method,the two diagnostic methods for the same cell has a one-to-one relationship to facilitate the doctor to make accurate judgments,with high efficiency,accurate and fast features.(2)Developed two different principles of production machine for the production requirements and market demand.Firstly,aiming at the shortcomings of the existing product structure,the low utilization rate of the manipulator and the low efficiency of cell collection,the polarized manipulator and the new work plan were designed to improve the degree of automation.At the same time,a new dynamic sealing operation head and pressure control system were designed to improve the cell acquisition effect,experimental results show that the production of the program is compact,stable and with high achievement satisfaction.Secondly,for the sedimentation-based liquid-based cell preparation machine,the commonly used specimen transfer machine function in the market was incorporated into it to achieve the pipetting and production automation and costly and more wasteful disposable syringes were replaced by costly pipettes to reduce the cost of supplies.Moreover,aiming at the problem of low efficiency and chaos of transfer specimens,a gantry truss three-dimensional manipulator and a folding centrifuge tube transfer frame were designed,a set of optimized motion control algorithm was explored on the basis of single chip to achieve a flexible arm of the linear interpolation and acceleration and deceleration movement and to improve work efficency.The circular interpolation also improved the tip clogging.Both actual test arm positioning accuracy and acceleration and deceleration curve fitting have high precision.(3)Developed an automatic dyeing machine to meet the demand of compound dyeing.In order to overcome the shortcomings of the existing dyeing machine,such as high price,large volume or poor corrosion resistance,the rotation scheme was chosen as the structural basis,the cooperation scheme of the multiplexed cylinder head and the master and slave robot was developed,which greatly improved the space utilization rate.From the material and structural design of high volatility and permeability of the strong acid for effective protection,put an end to the corrosion phenomenon;To improve the working efficiency,the dyeing process segmention algorthm was designed and achieved multi-tasking multiple dyeing at the same time.The results show that the accuracy of dyeing,repeatability,temperature control effect and the actual dyeing effect can meet the actual needs of the hospital and the needs of the project.They also show that the designe dyeing machine has the advantages of low cost,high efficiency,compact structure and excellent corrosion resistance.(4)Established a set of multi-band light source microspectroscopic imaging system and be able to stripping of the aliased multispectral image with mutiple linear regression method,completed DNA quantification and cell morphology synchronization analysis.The design of several groups can be issued discrete monochromatic LED combination of light source to reduce the cost and improve the stability of the instrument.Moreover,to improve the image accuracy and signal to noise ratio,a higher resolution and frame acquisition speed CMOS image sensor was used to obtain the spectral image;In the image transmission and processing methods,we used standard PCIe bus interface to transmit data and cached the image with DDR to enhance the image transmission speed to meet the real-time requirements;Finally,we used the multivariate linear regression method to isolate the true absorbance of the Fulgen dyes from the aliased multispectral images for quantitative analysis of DNA and to synthesize color images that meet the diagnostic criteria and requirements of the physician.The results show that it is feasible to choose a multi-spectral light imaging system with multi-band light source.As long as the LED of the appropriate band is selected,it can be easily realized in both optical structure and imaging method.It has the advantages of low cost,simple realization and good scalability specialty.Finally,the entire process was run in a continuous manner.In an experiment,we use the preparation of this paper machine,dyeing machine and microscopic spectral imaging of DNA quantitative analysis system to deal with and analysis of mouse liver and cervical specimens and use data measured by calibrated optical density test equipment as a comparison.The test results show that:the specimen preparation device designed in this paper can produce satisfactory smear and can successfully compound staining,using microscopic spectral analysis system to calculate the DNA accuracy is high,the method in the screening of cancer cells are high The accuracy and reliability,with great potential for market applications.