On-Chip Stimulated Raman Scattering Imaging and Quantification of Molecular Diffusion in Aqueous Microfluidics

Numerous chemical reactions and most life processes occur in aqueous solutions, where the physical diffusion of small molecules plays a vital role, including solvent water molecules, solute biomolecules, and ions. Conventional methods of measuring diffusion coefficients are often limited by technical complexity, large sample consumption, or significant time cost. Here, we present an optical imaging method to study molecular diffusion by combining stimulated Raman scattering (SRS) microscopy with microfluidics: a "Y"-shaped microfluidic channel forming two laminar flows with a stable concentration gradient across the interface. SRS imaging of a specific molecule allows us to obtain a high-resolution chemical profile of the diffusion region at varying inspection locations and flow rates, which enables the extraction of diffusion coefficients using the convection-diffusion model. As a proof of concept, we measured diffusion coefficients of molecules including water, protein, and multiple ions, with a sample volume of less than 1 mL and a time cost of less than 10 min. Moreover, we demonstrated a high-resolution three-dimensional (3D) reconstruction of the diffusion patterns in the microfluidic channel. The high-speed microfluidic SRS platform holds the potential for quantitative measurements of molecular diffusion, chemical reaction, and fluidic dynamics at the liquid-liquid interfaces.