microfluidic pressure regulator  (Elveflow Inc)

Journal: Advanced Science

Article Title: Multiplex Digital Methylation‐Specific PCR for Noninvasive Screening of Lung Cancer

doi: 10.1002/advs.202206518

Figure Lengend Snippet: Overview of mdMSP for noninvasive screening for early‐stage lung cancer. DNA is extracted from liquid biopsy specimens derived from CT‐scan positive patients. Following bisulfite conversion, the DNA sample is loaded into one module of a microfluidic chip. Digital PCR is performed on a flatbed heater in the presence of target‐specific, fluorescently‐labeled TaqMan probes to achieve simultaneous detection of four DNA targets. The microfluidic device is imaged with a standard flatbed imager, which is equipped with combinations of lasers and filters for the four fluorophores, FAM, HEX, Texas Red and Cy5. Last, the resulting images are analyzed using our customized software program to evaluate the intensity of the fluorescence generated from the four TaqMan fluorophores. The total number of detected copies of each biomarker per mL of plasma is summarized into a histogram to determine appropriate thresholds for optimizing assay performance. The dashed lines represent individual thresholds, which were selected based on desired sensitivity and specificity. Some elements in this figure were created with BioRender.com.

Article Snippet: After PCR was completed, the microfluidic chip was disconnected from the pressure regulator, removed from the thermal cycler, and then scanned with an Amersham Typhoon 5 Biomolecular Imager (GE Healthcare).

Techniques: Derivative Assay, Computed Tomography, Digital PCR, Labeling, Software, Fluorescence, Generated, Biomarker Assay