Multiple exosome RNA analysis methods for lung cancer diagnosis through integrated on-chip microfluidic system

Exosomes are now raising focus as a prospective biomarker for cancer diagnostics and prognosis owing to its unique bio-origin and composition.Exosomes take part in cellular communication and receptor mediation and transfer their cargos(e.g.,proteins,m RNA and DNA).Quantitative analysis of tumor-related nucleic acid mutations can be a potential method to cancer diagnosis and prognosis in early stages.Here we present an integrated microfluidic system for exosome on-chip isolation and lung cancer RNA analysis through droplet digital PCR(dd PCR).Gradient dilution experiments show great linearity over a large concentration range with R^(2)=0.9998.Utilizing the system,four cell lines and two mutation targets were parallelly detected for mutation analysis.The experiments demonstrated mutation heterogeneity and the results were agree with cell researches.These results proved our integrated microfluidic system as a promising means for early cancer diagnosis and prognosis in the era of liquid biopsy.

Single-cell level point mutation analysis of circulating tumor cells through droplet microfluidics

Tumor heterogeneity plays a critical role in the determination of appropriate anticancer therapy.As cir-culating tumor cells(CTCs)contain all tumor-related information,the genetic changes on CTCs could help us choose the appropriate treatments for different patients.Single-base mutations are very common in tumor genetic changes which may result in drug resistance.Here,we introduce a single-cell mutation de-tection platform based on droplet microfluidics.This platform integrates cell capsulation,cell lysis,poly-merase chain reaction(PCR)and the observation process.The droplets’generation speed is over 6000 per minute and more than 600 cells could be encapsulated in one second.To verify the performance of our platform in practical use,we performed the mutation analysis of 4 kinds of cells with our platform and noted that the genetic status of each single cell was clearly discriminated.Moreover,these results agreed with those from direct sequencing.Compared with other forms of single-cell mutation detection techniques,our platform has high throughput,short experimental time and less experimental operations.