Extending the dynamic range of biomarker quantification through molecular equalization.

Precision medicine requires highly scalable methods of multiplexed biomarker quantification that can accurately describe patient physiology. Unfortunately, contemporary molecular detection methods are generally limited to a dynamic range of sensitivity spanning just 3-4 orders of magnitude, whereas the actual physiological dynamic range of the human plasma proteome spans more than 10 orders of magnitude. Current methods rely on sample splitting and differential dilution to compensate for this mismatch, but such measures greatly limit the reproducibility and scalability that can be achieved-in particular, the effects of non-linear dilution can greatly confound the analysis of multiplexed assays. We describe here a two-pronged strategy for equalizing the signal generated by each analyte in a multiplexed panel, thereby enabling simultaneous quantification of targets spanning a wide range of concentrations. We apply our 'EVROS' strategy to a proximity ligation assay and demonstrate simultaneous quantification of four analytes present at concentrations spanning from low femtomolar to mid-nanomolar levels. In this initial demonstration, we achieve a dynamic range spanning seven orders of magnitude in a single 5 µl sample of undiluted human serum, highlighting the opportunity to achieve sensitive, accurate detection of diverse analytes in a highly multiplexed fashion.

MEMS 3D Scan Mirror with SU-8 Membrane and Flexures for High NA Microscopy.

We demonstrate a MEMS beam scanner capable of biaxial scanning with simultaneous focus control, for integration into a handheld confocal microscope for skin imaging. The device is based on a dual axis gimbal structure with an integrated largestroke deformable mirror. SU-8 polymer is used to construct both the deformable membrane as well as the torsional hinges for biaxial scanning. The 4 mm diameter mirror can perform raster pattern scanning with a range of +/- 1.5 degrees and Lissajous scanning with a range of +/- 3 degrees (mechanical scan angle), and has a maximum deflection of 9 um for focus control. The design, fabrication and characterization of the opto-mechanical performance of the MEMS device are presented in this paper.