Monolayer-Fluorescence Counting For The Ultrasensitive Detection Of Tumor-Derived Exosomes On An Acoustic Waves And Microstructure Assisted Microfluidic Chip

Oral cancer,as a common malignant tumor,is not easily detected due to its long latent period,and patients are often in the middle or late stage of the disease when they are clinically diagnosed,so the survival rate of patients after treatment is poor.If Early diagnosis of oral cancer can be achieved,it can effectively prolong the life of patients.Therefore,the search for a marker that can be used for early diagnosis of oral cancer has become the key of current research.Previous studies have shown that tumor-derived exosomes participate in the exchange of genetic information between tumor cells,thereby promoting the growth and metastasis of tumor cells.By detecting biomarkers such as exosomes,early diagnosis,precise treatment,realtime monitoring and prognosis of cancer can be realized.Compared with traditional methods,the detection of exosomes based on microfluidic technology has the advantages of fast diagnosis,less consumption of patient samples and reagents,high portability,and high detection reproducibility.In traditional microbiological methods,the collection,cultivation,and amplification of microorganisms are performed separately from analysis and identification,while in microfluidic systems,microfluidic chips can combine all these independent functions into a complex system to achieve simple,rapid and highly specific detection.In this study,a highly integrated and sensitive method for the detection of exosomes secreted by oral cancer cells CAL-27 was established by combining the morphological characteristics of exosomes and the specific proteins on its surface.Among them,the acoustic microfluidic chip is used to realize the efficient mixing of the fluid in the microchannel and improve the reaction efficiency;the stepped micro-nano structure microfluidic chip realizes the monolayer arrangement of the captured exosomes in the detection area,and the exosome in this area uses the fluorescence counting strategy to achieve high sensitive detection,due to its ability to eliminate background fluorescence interference and has a high signal-to-noise ratio.At the same time,an integrated experimental device that cooperates with the microfluidic chip is built,and the automatic sampling of the detection process is realized through the human-computer interaction of the serial screen.Other hardware devices were integrated into the acrylic preparation experimental box in an appropriate layout according to the function and wiring,thus achieving the integration and portability of the detection device.