Lung Cancer Cell Extraction And Single Cell Analysis In Sputum Are Used For Early Diagnosis And Histological Classification Of Lung Cancer

Among all of the cancers,the incidence and mortality of lung cancer have increased significantly in the world,which rank in the the forefront place of malignant tumors.The onset of lung cancer is usually concealed,and most lung cancers are diagnosed at an advanced stage.Statistics show that the 5year survival rate is only about 10%.Therefore,early detection of lung cancer is of great significance.Nonsmall cell lung cancer(NSCLC)accounts for approximately 80% of all lung cancers.adenocarcinoma(ADC)and squamous cell carcinoma(SqCC)are the main histological subtypes.Accurate confirmation of subtypes of lung cancer is a prerequisite for personalized diagnosis and treatment of lung cancer.So far,there are many methods for clinical diagnosis of lung cancer.Among them,pathological diagnosis is considered to be the most valuable for lung cancer confirmation.However,this method is usually used after the diagnosis of highly suspected cases by imaging and marker detection.Most of the pathological tissue samples need to be obtained by surgery or puncture,which is less meaningful for early detection of lung cancer and highrisk population screening.Sputum is a secretion of the lungs and tracheal respiratory tract.Studies have shown that cancer cells can already be detected in sputum before solid tumor formation.Therefore,exfoliated cells in sputum are ideal samples for pathological detection instead of tissues,and noninvasive pathological analysis of lung cancer can be achieved by obtaining cells for immunoassay.As a secretion of the lung,bronchus,and tracheal respiratory tract,the components in the sputum are complex,the cell types are diverse,and the number of tumor cells is small.Therefore,it is a challenge to sort and acquire a small amount of tumor cells from the sputum sample and analyze it.Microfluidic technology has the characteristics of controllability,high efficiency and easy integration.It has unique advantages in small amount of cell analysis.In addition,microfluidic chip has low manufacturing cost and easy operation,which is more conducive to the popularization of clinical applications.Therefore,microfluidic systems can be used to efficiently capture single tumor cells and perform in situ analysis on the chip to achieve accurate diagnosis of lung cancer.Based on this,combined with the advantages of microfluidic technology in analyzing a small number of cell samples and the characteristics of lung cancer as a respiratory disease,we use the microfluidic system to sort and capture lung cancer cells from sputum and analyze cells with multiplex immunofluorescence.It is intended to solve the problems of low adaptability caused by invasive sampling of traditional histopathology and poor recognition,low efficiency and difficult to automate operation of traditional sputum cytology detection,which is provide new strategies and approaches for early non-invasive detection of lung cancer and screening of practical lung cancer in high-risk groups.The first chapter is the introduction.We first expound the research status and significance of early diagnosis of lung cancer and lung cancer typing.Then we summarize the existing methods of capturing and analyzing single cells based on microfluidic system.we use the sputum exfoliated cells for the feasibility and innovation of lung cancer diagnosis.We demonstrate the feasibility and innovation of using microfluidic systems to diagnose lung cancer with cells from sputum.The second chapter is the design and construction of a microfluidic system for sorting and capturing tumor single cells from sputum.For the first time,we proposed the use of microfluidic system to separate and immunologically classify lung cancer cells in sputum for accurate diagnosis of lung cancer.In this chapter,we design and use spiral chips and single-cell array chips to capture single-cell and online multiplex immunofluorescence analysis of tumor cells in sputum.We use a small number of non-invasive cell samples to replace traditional tissue immunohistochemistry,which is expected to achieve non-invasive accurate diagnosis and classification of lung cancer,It provides a new idea for the treatment and prognosis monitoring of early stage lung cancer patients,high-risk groups and advanced patients.At the same time,the singlecell immunofluorescence technology on the chip provides a good reference for similar research.The third chapter is the surface of the cell modified immunofluorescence microsphere to assis the capture and analysis of single cells.Single-cell array chip is the chip with capabilities of cell capture and online analytical,which offer unique advantages in the processing of small numbers of cell samples.However,the characteristics of physical sorting determine the chip having the problem of selectivity and sorting speed.Therefore,we combine microsphere immunocapture technology with single-cell array chip technology to achieve high-speed,high-selective capture and analysis of the chip.First of all,we use microsphere immunocapture to specifically recognize tumor cells,avoiding the interference of cells which with the similar size.At the same time,the surface of the cell is modified with microspheres to reduce the cell deformability so that the cells can be firmly trapped at the micro chamber under a higher flow rate,which greatly improve the capture flow rate.Finally,the unique in-situ analysis performance of single cell array chip can be used for in-situ analy of the captured tumor cells,avoiding the false-positive problem.