Two-Dimensional Light Scattering Flow Cytometry With Multiple Sheath Flows And Its Application In Lung Cancer Detection

The morbidity and mortality of lung cancer are the highest in China and throughout the world.According to the treatment and biological characteristics,lung cancer is differentiated into small cell lung cancer(SCLC)and non-small cell lung cancer(NSCLC).At present,the diagnosis of lung cancer mainly depends on tissue biopsy,which requires high experience of physicians,complicated sampling and staining procedures,and is subjective.Conventional techniques differ in distinguishing SCLC and NSCLC.Therefore,it is of great medical value to develop a new diagnostic method for lung cancer.Label-free two-dimensional(2D)light scattering technology has unique advantages in the field of cell analysis.It can be used for single-cell high-throughput analysis combining flow technology.With the development of micromachining technology,miniaturized microfluidic cytometer has become an important direction of flow cytometry.In order to further improve the flux of flow cytometry and realize parallel detection,multi-channel flow cytometry has gradually attracted people’s interest.The combination of 2D light scattering technology and multi-channel sheath flow technology is expected to realize high-throughput and label-free detection of lung cancer.In this paper,a 2D light scattering flow cytometer with multiple sheath flows is constructed by integrating the light sheet technology,multi-channel three-dimensional hydrodynamic focusing technology,and label-free 2D light scattering technology.The device mainly includes three modules:light sheet generation module,multi-channel 3D hydrodynamic focusing module and scattering pattern acquisition and processing module.The light sheet generation module compresses the laser beam of the spindle-shaped cross section into a light sheet.The function of the multi-channel 3D hydrodynamic focusing module is to simultaneously form multiple independent focusing streams to improve the detection flux.The scattering pattern acquisition and processing module collects 2D light scattering patterns of particles or cells and analyzes them.Then,2D light scattering patterns of small cell lung cancer cell line(H69)cells and human poorly differentiated lung adenocarcinoma cell line(SK-LU-1)cells are collected.With the help of the four features extracted by the gray level co-occurrence matrix,this paper realizes the automatic classification of the two types of lung cancer cells through the SVM classifier,and the classification accuracy rate reaches 92.75%.The results of this study show that 2D light scattering flow cytometry with multiple sheath flows has the advantages of label-free and high-throughput.It is expected to provide a new detection method for the diagnosis of lung cancer and other cancers.