There have been growing interests in droplet-based microfluidics due to its capability to outperform conventional biological assays by providing various advantages, such as precise handling of liquid/cell samples, fast reaction time, and extremely high-throughput analysis/screening. The droplet-based microfluidics utilizes the interaction between the interfacial tension and the fluidic shear force to break continuous fluids into uniform-sized segments within a microchannel. In this paper, the effect of different viscosities of carrier oil on water-in-oil emulsion, particularly how droplet size and droplet generation rate are affected, has been investigated using a commonly used T-junction microfluidic droplet generator design connected to a pressure-controlled pump. We have tested mineral oils with four different viscosities (5, 7, 10, and 15 cSt) to compare the droplet generation under five different flow pressure conditions (i.e., water flow pressure of 30–150 mbar and oil flow pressure of 40–200 mbar). The results showed that regardless of the flow pressure levels, the droplet size decreased as the oil viscosity increased. Average size of the droplets decreased by approximately 32% when the viscosity of the oil changed from 5 to 15 cSt at the flow pressure of 30 mbar for water and 40 mbar for oil. Interestingly, a similar trend was observed in the droplet generation rate. Droplet generation rate and the oil viscosity showed high linear correlation (R² = 0.9979) at the water flow pressure 30 mbar and oil flow pressure 40 mbar.

Polydimethylsiloxane (PDMS) is a polymer widely used for fabrication and prototyping of microfluidic chips. The porous matrix structure of PDMS allows small hydrophobic molecules including some fluorescent dyes to be readily absorbed to PDMS and results in high fluorescent background signals, thereby significantly decreasing the optical detection sensitivity. This makes it challenging to accurately detect the fluorescent signals from samples using PDMS devices. Here, we have utilized polytetrafluoroethylene (PTFE) to inhibit absorption of hydrophobic small molecules on PDMS. Nile red was used to analyze the effectiveness of the inhibition and the absorbed fluorescence intensities for 3% and 6% PTFE coating (7.7 ± 1.0 and 6.6 ± 0.2) was twofold lower compared to 1% and 2% PTFE coating results (17.2 ± 0.5 and 15.4 ± 0.5). When compared to the control (55.3 ± 1.6), it was sevenfold lower in background fluorescent intensity. Furthermore, we validated the optimized PTFE coating condition using a PDMS bioreactor capable of locally stimulating cells during culture to quantitatively analyze the lipid production using Chlamydomonas reinhardtii CC-125. Three percent PTFE coating was selected as the optimal concentration as there was no significant difference between 3% and 6% PTFE coating. Intracellular lipid contents of the cells were successfully stained with Nile Red inside the bioreactor and 3% PTFE coating successfully minimized the background fluorescence noise, allowing strong optical lipid signal to be detected within the PDMS bioreactor comparable to that of off-chip, less than 1% difference.