Simultaneous Absorbance and Fluorescence Measurements Using an Inlaid Microfluidic Approach

A novel microfluidic optical cell is presented that enables simultaneous measurement of both light absorbance and fluorescence on microlitre volumes of fluid. The chip design is based on an inlaid fabrication technique using clear and opaque poly(methyl methacrylate) or PMMA to create a 20.2 mm long optical cell. The inlaid approach allows fluid interrogation with minimal interference from external light over centimeter long path lengths. The performance of the optical cell is evaluated using a stable fluorescent dye: rhodamine B. Excellent linear relationships (R² > 0.99) are found for both absorbance and fluorescence over a 0.1-10 µM concentration range. Furthermore, the molar attenuation spectrum is accurately measured over the range 460-550 nm. The approach presented here is applicable to numerous colorimetric- or fluorescence-based assays and presents an important step in the development of multipurpose lab-on-chip sensors.