A microchip electrophoresis method was developed for simultaneous determination of reactive oxygen species (ROS) and reduced glutathione (GSH) in the individual erythrocyte cell. In this method, cell sampling, single-cell loading, docking, lysing, and capillary electrophoretic separation with LIF detection were integrated on a microfluidic chip with crossed channels. ROS was labeled with dihydrorhodamine 123 in the intact cell, while GSH was on-chip labeled with 2,3-naphthalene-dicarboxaldehyde, which was included in the separation medium. On-chip electrical lysis, characterized by extremely fast disruption of the cellular membrane (<40 ms), was exploited to minimize enzymatic effects on analyte concentrations during the determination. The microfluidic network was optimized to prevent cell leaking from the sample reservoir (S) into separation during the separation phase. The structure of the S was modified to avoid blockage of its outlet by deposited cells. Detection limits of 0.5 and 6.9 amol for ROS and GSH, respectively, were achieved. The average cell throughput was 25 cells/h. The effectiveness of the method was demonstrated in the simultaneous determination of GSH and ROS in individual cells and the variations of cellular GSH and ROS contents in response to external stimuli.

Reactive oxygen species (ROS) are known to not only mediate the damage of cellular constituents but also to regulate cellular signaling. Analysis of ROS is essential if we wish to understand the mechanisms of cellular alterations. In this paper, a microfluidic chip-based approach to the determination of ROS in single erythrocyte was developed by using a simple crossed-channel glass chip with integrated operational functions, including cell sampling, single cell loading, docking, lysing, and capillary electrophoretic (CE) separation with laser-induced fluorescence (LIF) detection. Non-fluorescent dihydrorhodamine 123 (DHR 123), which can be oxidized intracellularly by ROS to the fluorescent rhodamine 123 (Rh 123), was used as the fluorogenic reagent. The effect of pH on the migration time of Rh 123 and detection sensitivity was discussed. The present method minimized dilution of intracellular ROS during reaction with DHR 123 and determination. As a result, an extremely low detection limit of 0.8 amol has been achieved. The time required for complete analysis of one human erythrocyte was less than 3 min. A migration time precision of 4.1% RSD was obtained for six consecutively-injected cells. Upon stimulation with 4 mmol/l H2O2 for 10 min, the intracellular ROS concentration was found to increase on average by about a factor of 8.4.

Ultrathin silica coating (UTSC) has emerged as an effective way to improve the compatibility and stability of nanoparticles without attenuating their intrinsic optical properties. Exploration strategies to probe the growth process of ultrathin silica shells on the surface of nanoparticles would represent a valuable innovation that would benefit the development of ultrathin silica coated nanoparticles and their relevant applications. In this work, we report a unique, very effective and straightforward strategy for probing the growth of ultrathin silica shells on the surface of gold nanorods (Au NRs), which exploits the localized surface plasmon resonance (LSPR) as a reporting signal. The thickness of the ultrathin silica shells on the surface of Au NRs can be quantitatively measured and predicted in the range of 0.5-3.5 nm. It is demonstrated that the LSPR shift accurately reflects the real-time change in the thickness of the ultrathin silica shells on Au NRs during the growth process. By using the developed strategy, we further analyze the growth of UTSC on the surface of Au NRs via feeding of Na2SiO3 in a stepwise manner. The responsiveness analysis of LSPR also provides important insight into the shielding effect of UTSC on the surface of Au NRs that is not accessible with conventional strategies. This LSPR-based strategy permits exploration of the surface-mediated sol-gel reactions of silica from a new point of view.

POEGMA-based disulfide-containing fluorescent probes were designed and synthesized to enable the real-time and quantitative analysis of the (bio)reduction of disulfides within the polymeric backbones in complex redox media and in the presence of cells. This study lights up the way of exploiting bioreduction in the extracellular spaces for the delivery of hydrophobic drugs.

We present the fabrication and optimization of new integrated microgiant electrorheological ͑GER͒ valves for microflow cytometry. Compared to previous GER valves, new GER valves, consisting of an SU-8 layer, a PDMS membrane, and glass layers, were fabricated by 4-mask microelectromechanical systems technology and two new packaging methods. This design enabled good bonding and fluidic interconnection. The thickness of the PDMS membrane was designed such that the membrane deformation was large enough that the cytometry channel was well sealed. The interfacial stress between the PDMS and the PDMS/SU-8 as a function of vacuum plasma treatment time was investigated in detail. The switching behavior of the GER valves was also analyzed and characterized using fluorescence microscopy.