Optimizing Microfluidic Impedance Cytometry by Bypass Electrode Layout Design.

Microfluidic impedance cytometry (MIC) has emerged as a popular technique for single-cell analysis. Traditional MIC electrode designs consist of a pair of (or three) working electrodes, and their detection performance needs further improvements for microorganisms. In this study, we designed an 8-electrode MIC device in which the center pair was defined as the working electrode, and the connection status of bypass electrodes could be changed. This allowed us to compare the performance of layouts with no bypasses and those with floating or grounding electrodes by simulation and experiment. The results of detecting Φ 5 µm beads revealed that both the grounding and the floating electrode outperformed the no bypass electrode, and the grounding electrode demonstrated the best signal-to-noise ratio (SNR), coefficient of variation (CV), and detection sensitivity. Furthermore, the effects of different bypass grounding areas (numbers of grounding electrodes) were investigated. Finally, particles passing at high horizontal positions can be detected, and Φ 1 µm beads can be measured in a wide channel (150 µm) using a fully grounding electrode, with the sensitivity of bead volume detection reaching 0.00097%. This provides a general MIC electrode optimization technology for detecting smaller particles, even macromolecular proteins, viruses, and exosomes in the future.

Direct, ultrafast, and sensitive detection of environmental pathogenic microorganisms based on a graphene biosensor.

Pathogenic microorganisms in the environment pose a serious threat to global human health. This study developed a reduced graphene oxide (rGO)-field effect transistor (FET) biosensor to realize the rapid and sensitive detection of pathogenic microorganisms. The rGO-FET sensors were prepared by in-situ thermal reduction method, and biorecognition elements were immobilized using a crosslinking agent to realize the surface functionalization of rGO. The rGO-FET biosensors can detect Escherichia coli O157:H7 as low as 1.4 CFU mL-1 within 46 s. The normalized current response was linearly correlated with E. coli concentration in the range of 1.4-1.4 × 107 CFU mL-1. The normalized current response of E. coli O157:H7 was about an order of magnitude higher than those of other microorganisms, indicating that the biosensor has good specificity. The current loss rates of the unmodified rGO-FET sensors and the biosensors modified with anti-E. coli O157:H7 after 30 days of storage at 4 °C were approximately 8% and 15%, respectively. Most importantly, the rGO-FET biosensors can directly detect real samples without pretreatment. Compared with other technologies, the rGO-FET biosensors can detect pathogenic microorganisms with a wider linear range in a shorter time, which is of great importance for the rapid warning and control of pathogenic microorganisms in the environment.