Mussel-inspired enzyme immobilization and dual real-time compensation algorithms for durable and accurate continuous glucose monitoring.

Blood glucose sensing is very important for diabetic management. It is shifting towards a continuous glucose monitoring because such a system can alleviate patient suffering and provide a large number of glucose measurements. Here, we proposed a novel approach for the development of durable and accurate enzymatic continuous glucose monitoring system (CGMS). For the long-term durable and selective immobilization of glucose oxidase on a microneedle electrode, a biocompatible engineered mussel adhesive protein was employed through efficient electrochemical oxidation strategy. For the accurate performance in in vivo environments, we also suggested dual real-time compensated algorithms to cover both temperature and time-lag differences. After pre-clinical and pilot-clinical evaluations, we confirmed that our proposed CGMS has an outstanding performance compared with various commercially available continuous systems and achieves comparable performance to disposable glucose sensors.

Glutaminase 2 expression is associated with regional heterogeneity of 5-aminolevulinic acid fluorescence in glioblastoma.

Fluorescence-guided surgery using 5-aminolevulinic acid (5-ALA) is now a widely-used modality for glioblastoma (GBM) treatment. However, intratumoral heterogeneity of fluorescence intensity may reflect different onco-metabolic programs. Here, we investigated the metabolic mechanism underlying the heterogeneity of 5-ALA fluorescence in GBM. Using an in-house developed fluorescence quantification system for tumor tissues, we collected 3 types of GBM tissues on the basis of their fluorescence intensity, which was characterized as strong, weak, and none. Expression profiling by RNA-sequencing revealed 77 genes with a proportional relationship and 509 genes with an inverse relationship between gene expression and fluorescence intensity. Functional analysis and in vitro experiments confirmed glutaminase 2 (GLS2) as a key gene associated with the fluorescence heterogeneity. Subsequent metabolite profiling discovered that insufficient NADPH due to GLS2 underexpression was responsible for the delayed metabolism of 5-ALA and accumulation of protoporphyrin IX (PpIX) in the high fluorescence area. The expression level of GLS2 and related NADPH production capacity is associated with the regional heterogeneity of 5-ALA fluorescence in GBM.

Nonfaradaic nanoporous electrochemistry for conductometry at high electrolyte concentration.

Nanoporous electrified surfaces create a unique nonfaradaic electrochemical behavior that is sensitively influenced by pore size, morphology, ionic strength, and electric field modulation. Here, we report the contributions of ion concentration and applied ac frequency to the electrode impedance through an electrical double layer overlap and ion transport along the nanopores. Nanoporous Pt with uniform pore size and geometry (L2-ePt) responded more sensitively to conductivity changes in aqueous solutions than Pt black with poor uniformity despite similar real surface areas and enabled the previously difficult quantitative conductometry measurements at high electrolyte concentrations. The nanopores of L2-ePt were more effective in reducing the electrode impedance and exhibited superior linear responses to not only flat Pt but also Pt black, leading to successful conductometric detection in ion chromatography without ion suppressors and at high ionic strengths.

A flow cytometry-based submicron-sized bacterial detection system using a movable virtual wall.

Detection of pathogenic bacteria requires a sensitive, accurate, rapid, and portable device. Given that lethal microbes are of various sizes, bacterial sensors based on DC (direct current) impedance on chips should be equipped with channels with commensurate cross sections. When it comes to counting and interrogation of individual bacteria on a microfluidic chip, very narrow channels are required, which are neither easy nor cost-effective to fabricate. Here, we report a flow cytometry-based submicron-sized bacterial detection system using a movable virtual wall made of a non-conducting fluid. We show that the effective dimension of a microfluidic channel can be adjusted by varying the respective flow rates of a sample solution as well as the liquid wall therein. Using such a virtual wall, we have successfully controlled the channel width and detected submicron-sized Francisella tularensis, a lethal, tularemia-causing bacterium. Since the system is capable of monitoring changes in DC impedance and fluorescence simultaneously, we were also able to discriminate between different types of bacterial mixtures containing F. tularensis and E. coli BL21 that have different gamuts of size distributions. The proposed flow cytometry-based system represents a promising way to detect bacteria including, but not limited to, submicron-sized pathogenic microbes.

A label-free DC impedance-based microcytometer for circulating rare cancer cell counting.

Quantification of circulating tumor cells (CTCs) in blood samples is believed to provide valuable evidence of cancer progression, cancer activity status, response to therapy in patients with metastatic cancer, and possible cancer diagnosis. Recently, a number of researchers reported that CTCs tend to lose their epithelial cell adhesion molecule (EpCAM) by an epithelial-mesenchymal transition (EMT). As such, label-free CTC detection methods are attracting worldwide attention. Here, we describe a label-free DC impedance-based microcytometer for CTCs by exploiting the difference in size between CTCs and blood cells. This system detects changes in DC impedance between two polyelectrolytic gel electrodes (PGEs) under low DC voltages. Using spiked ovarian cancer cell lines (OVCAR-3) in blood as a model system, we were able to count the cells using a microcytometer with 88% efficiency with a flow rate of 13 µl min(-1) without a dilution process. Furthermore, we examined blood samples from breast cancer patients using the cytometer, and detected CTCs in 24 out of 24 patient samples. Thus, the proposed DC impedance-based microcytometer presents a facile and fast way of CTC evaluation regardless of their biomarkers.