Journal: PLoS ONE

Article Title: A Microfluidic Platform for High-Throughput Multiplexed Protein Quantitation

doi: 10.1371/journal.pone.0117744

Figure Lengend Snippet: a) microfluidic design: the device consists of two PDMS layers: flow (blue) and control (red). The chip is an array of eight rows by 48 columns for 384 unit cells. Each unit cell is composed of: two antibody chambers divided by a reaction chamber (1–2), 4 MITOMI buttons (3), a valve that segregates the unit cells (4), a valve that separates antibody and reaction chambers (5) and a valve for releasing pressure in the antibody chambers (6). b) The PDMS chip is aligned to an epoxy-functionalized slide onto which primary and secondary antibodies were spotted. c) Assay details: schematic of the unit cell and cross section of a button region: i) functionalization of the surface: BSA-biotin is flowed though the chip followed by neutravidin. Next, the buttons are closed and BSA-biotin flowed again to passivate all neutravidin molecules except for those located underneath the MITOMI buttons, ii) the biotinylated primary antibody is allowed to diffuse into the MITOMI detection chamber and is bound by neutravidin immobilizing it in the MITOMI detection regions, iii) the sample is flown through the device and antigens are captured by the surface immobilized antibodies, iv) finally, the fluorescently labeled secondary antibody is allowed to diffuse into the MITOMI area, binds to the antigen if present, and is trapped by MITOMI. The entire device is then quantitated using a DNA microarray scanner.

Article Snippet: The microfluidic device was scanned using a fluorescent microarray scanner (ArrayWorx e-Biochip Reader, Applied Precision, USA).

Techniques: Control, Labeling, Microarray