Design Of A Detection Device For Circulating Tumor Cells As Environmental Carcinogenic Markers And Their Detection Imaging

With the acceleration of industrialization,environmental pollution has become increasingly serious,and the rapid accumulation of carcinogen in the environment has led to an increase in the incidence rate of environmental cancer year by year.Research shows that only about 5% of cancers are caused solely by genetic factors,while 80% of cancers can be attributed to environmental factors.Early prediction and diagnosis of cancer require detection of biomarkers,including serum markers,gene markers,and cell markers.Among the cell markers,circulating tumor cells(CTC)are now proven reliable early markers for cancer patients.Therefore,the detection of CTC can be effectively applied to the early diagnosis of tumors and evaluate the therapeutic effect.It can be used as an "environmental carcinogenic marker" for health monitoring of environmental carcinogenic populations.Based on this,we conducted research on the design of detection devices and imaging detection of circulating tumor cells as environmental carcinogenic markers,providing detection devices and methods for the detection of environmental carcinogenic marker CTC.Firstly,construct a specific recognition method for tumor cell imaging detection.Using human breast cancer cell MCF-7 with high expression of Ep CAM as the cell model,Ep CAM was specifically recognized by using FAM labeled SYL-3C aptamer.The results indicate that under blood interference,the aptamer can specifically recognize MCF-7 cells.In order to detect specific recognition fluorescence under inverted fluorescence microscopy and achieve visualization of cell contour,a 13mol/L Tween-80 solution of N,N-dimethylformamide was used to enhance the specific recognition fluorescence.When the fluorescence signal-to-noise ratio on the surface of tumor cells increases,combined with digital signal amplification technology,the imaging contour of the cells is presented completely and clearly,avoiding signal loss in imaging cells,providing a complete and clear imaging signal for accurate counting and scale estimation of CTC in tumor cell processors.Secondly,in order to avoid cell loss caused by centrifugation during cell preprocessing before imaging,an integrated filtering separation and imaging detection device was designed and processed,which includes pre processing functions before cell detection and imaging detection functions after processing cells,abbreviated as the tumor cell detection processor.To prevent cell fragmentation caused by external forces during filtration,a negative pressure free filtration method is adopted.Using MCF-7 cells as model cells,specific recognition was performed using fluorescein labeled Ep CAM nucleic acid aptamer Apt-FAM.Combined with fluorescence enhancement and digital signal amplification techniques,the performance of the tumor cell detection processor was evaluated.The results show that the processor can achieve imaging detection of CTC,with clear imaging and ensuring the accuracy of cell counting.Subsequently,to verify whether the cell processor is suitable for antibody detection of tumor cells,rabbit anti human Ep CAM antibodies and TRITC labeled donkey anti rabbit Ig G were used for imaging detection of Ep CAM overexpressing MCF-7.Research has shown that even under the interference of blood,target cells can be specifically identified.Combined with digital signal amplification technology,the cell contour is clear and complete,ensuring the accuracy of CTC counting.Therefore,the extremely low density tumor cell processor constructed in this article can achieve detection and imaging of captured cells,and the operation is simple,which is expected to be used for clinical detection of tumor cells.Finally,the tumor cell detection processor is used to capture samples for detection imaging.In order to allow the captured cells to detach from the trap and enter the processor without damage,a CTC pre-treatment device used in the supporting processor was designed and processed.At the same time,methods for ethanol cell fixation and cell detachment from the capture were constructed.Research has shown that after fixation,wash with PBS 12 times,100 times each time μ L can ensure that the captured cells on the filament completely fall off and enter the tumor cell processor.After shedding and imaging the captured cells,the target cells in the cell processor can be completely and clearly detected,verifying the effectiveness of the cell detection processor in detecting and imaging captured cells.