Single Escherichia coli bacteria detection using a chemiluminescence digital microwell array chip.

Rapid and sensitive Escherichia coli (E. coli) detection is important in determining environmental contamination, food contamination, as well as bacterial infection. Conventional methods based on bacterial culture suffer from long testing time (24 h), whereas novel nucleic acid-based and immunolabelling approaches are hindered by complicated operation, the need of complex and costly equipment, and the lack of differentiation of live and dead bacteria. Herein, we propose a chemiluminescence digital microwell array chip based on the hydrolysis of 6-Chloro-4-methylumbelliferyl-ß-D-glucuronide by the ß-D-glucuronidase in E. coli to achieve fast single bacterial fluorescence detection. Taking the advantage of the picoliter microwells, single bacteria are digitally encapsulated in these microwells, thus the accurate quantification of E. coli can be realized by counting the number of positive microwells. We also show that the chemiluminescence digital microwell array chip is not affected by the turbidity of the test samples as well as the temperature. Most importantly, our method can differentiate live and dead bacteria through bacterial proliferation and enzyme expression, which is confirmed by detecting E. coli after pH and chlorination treatment. By comparing with the standard method of plate counting, our method has comparable performance but significantly reduces the testing time from over 24 h-2 h and 4 h for qualitative and quantitative analysis, respectively. In addition, the microfluidic chip is portable and easy to operate without external pump, which is promising as a rapid and on-site platform for single E. coli analysis in water and food monitoring, as well as infection diagnosis.

On-Chip Optical Detection of Viruses: A Review.

The current outbreak of the coronavirus disease-19 (COVID-19) pandemic worldwide has caused millions of fatalities and imposed a severe impact on our daily lives. Thus, the global healthcare system urgently calls for rapid, affordable, and reliable detection toolkits. Although the gold-standard nucleic acid amplification tests have been widely accepted and utilized, they are time-consuming and labor-intensive, which exceedingly hinder the mass detection in low-income populations, especially in developing countries. Recently, due to the blooming development of photonics, various optical chips have been developed to detect single viruses with the advantages of fast, label-free, affordable, and point of care deployment. Herein, optical approaches especially in three perspectives, e.g., flow-free optical methods, optofluidics, and surface-modification-assisted approaches, are summarized. The future development of on-chip optical-detection methods in the wave of emerging new ideas in nanophotonics is also briefly discussed.

Biotoxoid Photonic Sensors with Temperature Insensitivity Using a Cascade of Ring Resonator and Mach-Zehnder Interferometer.

The great advances in silicon photonic-sensing technology have made it an attractive platform for wide sensing applications. However, most silicon photonic-sensing platforms suffer from high susceptibility to the temperature fluctuation of an operating environment. Additional complex and costly chemical signal-enhancement strategies are usually required to improve the signal-to-noise ratio (SNR). Here, a biotoxoid photonic sensor that is resistant to temperature fluctuation has been demonstrated. This novel sensor consists of a ring resonator coupled to a Mach-Zehnder interferometer (MZI) readout unit. Instead of using costly wavelength interrogation, our photonic sensor directly measures the light intensity ratio between the two output ports of MZI. The temperature dependence (TD)-controlling section of the MZI is used to eliminate the adverse effects of ambient temperature fluctuation. The simulation and experimental results show a linear relationship between the interrogation function and the concentration of an analyte under operation conditions. The thermal drift of the proposed sensor is just 0.18%, which is a reduction of 567-fold for chemical sensing and 28-fold for immuno-biosensing compared to the conventional single-ring resonator. The SNR increases from 6.85 to 19.88 dB within a 2 °C temperature variation. The high SNR optical sensor promises great potential for amplification-free detection of nucleic acids and other biomarkers.

Nanophotonic Array-Induced Dynamic Behavior for Label-Free Shape-Selective Bacteria Sieving.

Current particle sorting methods such as microfluidics, acoustics, and optics focus on exploiting the differences in the mass, size, refractive index, or fluorescence staining. However, there exist formidable challenges for them to sort label-free submicron particles with similar volume and refractive index yet distinct shapes. In this work, we report an optofluidic nanophotonic sawtooth array (ONSA) that generates sawtooth-like light fields through light coupling, paving the physical foundation for shape-selective sieving. Submicron particles interact with the coupled hotspots which impose different optical torques on the particles according to their shapes. Unstained S. aureus and E. coli are used as a model system to demonstrate this shape-selective sorting mechanism based on the torque-induced body dynamics, which was previously unattainable by other particle sorting technologies. More than 95% of S. aureus is retained within ONSA, while more than 97% of E. coli is removed. This nanophotonic chip offers a paradigm shift in shape-selective sorting of submicron particles and expands the boundary of optofluidics-based particle manipulation.

Regulation of lipid droplets in live preadipocytes using optical diffraction tomography and Raman spectroscopy.

Lipid droplets have gained strong interest in recent years to comprehend how they function and coordinate with other parts of the cell. However, it remains challenging to study the regulation of lipid droplets in live preadipocytes using conventional microscopic techniques. In this paper, we study the effects of fatty acid stimulation and cell starvation on lipid droplets using optical diffraction tomography and Raman spectroscopy by measuring size, refractive index, volume, dry mass and degree of unsaturation. The increase of fatty acids causes an increase in the number and dry mass of lipid droplets. During starvation, the number of lipid droplets increases drastically, which are released to mitochondria to release energy. Studying lipid droplets under different chemical stimulations could help us understand the regulation of lipid droplets for metabolic disorders, such as obesity and diabetes.