Droplet microfluidics with push-pull and microdialysis sampling from brain slices, cultured cells and engineered tissues produce low volume mass limited samples containing analytes sampled from the extracellular space. This sampling approach coupled to mass spectrometry (MS) detection allows evaluation of time-dependent chemical changes. Our goal is an approach for continuous sampling and segregation of extracellular samples into picoliter droplets followed by the characterization of the droplets using nanoelectrospray ionization (nESI) MS. The main focus here is the optimization of the carrier oil for the microfluidic device that neither affects the stability of picoliter droplets nor compatibility with MS detection of a range of analytes.
Quantum dots (QDs)-based molecularly imprinted polymer (MIP) composite nanospheres were successfully prepared via a facile and versatile ultrasonication-assisted encapsulation method. Unlike the hydrogen-bond-based MIPs, these so-prepared QDs-MIP composite nanospheres, relying on the interaction including van der Waals forces and hydrophobic forces, demonstrated excellent selectivity in aqueous media. Their small particle sizes and carboxyl-enriched polymer matrixes give rise to their good dispersibility and stability in aqueous solution, and faster adsorption and desorption kinetics, which further make them extensively applicable for chemical/biological sensors in aqueous media. Based on the fluorescence quenching via template analytes (diazinon) rebinding into the recognition cavities in the polymer matrixes, the QDs-MIP nanospheres were successfully applied to the direct fluorescence quantification of diazinon, independent of extracting templates from the MIP nanospheres, as well as further complicated and time-consuming assays. This novel method can selectively and sensitively detect down to 50 ng/mL of diazinon in water, and a linear relationship has been obtained covering the concentration range of 50-600 ng/mL. The present studies provide a new and general strategy to fabricate other multifunctional (luminescent and magnetic) inorganic-organic MIP nanocomposites with highly selective recognition ability in aqueous media and are pretty desirable for biomedical/chemical sensing applications.
Abstract Integrating droplet-based microfluidics with mass spectrometry is essential to high-throughput and multiple analysis of single cells. Nevertheless, matrix effects such as the interference of culture medium and intracellular components influence the sensitivity and the accuracy of results in single-cell analysis. To resolve this problem, we developed a method that integrated droplet-based microextraction with single-cell mass spectrometry. Specific extraction solvent was used to selectively obtain intracellular components of interest and remove interference of other components. Using this method, UDP-Glc-NAc, GSH, GSSG, AMP, ADP and ATP were successfully detected in single MCF-7 cells. We also applied the method to study the change of unicellular metabolites in the biological process of dysfunctional oxidative phosphorylation. The method could not only realize matrix-free, selective and sensitive detection of metabolites in single cells, but also have the capability for reliable and high-throughput single-cell analysis.
The analysis of lipid droplets (LDs) by mass spectrometry at the single LD level is still an analytical challenge. In this work, we developed a novel technique termed in-tip solvent microextraction mass spectrometry for the separation and profiling of phosphatidylcholines and triglycerides within a single LD. This method has been successfully used to analyze LDs in mammalian cells and to compare the profiles of triglycerides and phosphatidylcholines in LDs induced at different conditions. Our method has the potential to be applied to such fields as fundamental lipid biology to further our understanding on the mechanisms of lipid production, lipid packaging, and their pathophysiological roles.
Patients with epidermal growth factor receptor mutant nonsmall cell lung cancer (NSCLC) often fail to treat gefitinib because of secondary drug resistance. The development of tumor drug resistance is closely related to variations in cancer cell metabolism. Single-cell metabolomics analysis can provide unique information about tumor drug resistance. Herein, we constructed a platform to study the secondary resistance of tumor cells based on single-cell metabolomics (sSRTC-scM). A gefitinib-resistant NSCLC cell line (PC9GR) was constructed by increasing the dose step by step. The metabolic profiles of parental PC9 cells and PC9GR cells with different drug resistance levels were detected by intact living-cell electrolaunching ionization mass spectrometry at the single-cell level. The data were analyzed by statistical methods such as t-SNE, variance, volcano plot, heat map, and metabolic pathway analysis. Using this platform, we found that the metabolic fingerprints of PC9GR cells can evaluate drug resistance degrees. The metabolic fingerprints continue to be altered with the increase of drug resistance. We revealed 19 metabolic markers of secondary resistance by variance analysis and clarified that the glycerophospholipid metabolic pathway of PC9GR cells changed significantly. In addition, we found that with the increase in drug resistance levels, the heterogeneity of single-cell metabolism became greater and the number of cells with weak drug resistance gradually decreased. This phenomenon can be utilized to illustrate the drug resistance degrees of PC9GR cells. This study provides diagnostic markers for evaluating the drug resistance of tumors and gives new insight into overcoming the secondary resistance of tumors.
Robotic-assisted surgery (RAS) is increasingly used for treating low rectal cancer. Its comparative effectiveness against laparoscopic surgery (LAS) in enhancing long-term anal function remains uncertain.
Current detection methods for paper-based analytical devices (PADs) rely on spectroscopic and electrochemical properties, which place special requirements on the analyte or need analyte labeling. Here, ion-transmission mass spectrometry (MS) was proposed for coupling with PADs to enable rapid in situ MS analysis of the sample on paper. The sample was analyzed directly on paper via analyte ionization by ions transmitted through the paper, generated by a low-temperature plasma probe. Prior to MS analysis, the sample can be separated by paper electrophoresis or by paper chromatography, among a variety of other features offered by PADs. The versatility of this technique was demonstrated by MS analysis of a paper microarray, a mixture of amino acids, and whole blood doped with drugs on PADs.
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