Intact living-cell electrolaunching ionization mass spectrometry for single-cell metabolomics

Shao, Yunlong; Zhou, Yang; Liu, Yuanxing; Zhang, Wenmei; Zhu, Guizhen; Zhao, Yaoyao; Zhang, Qi; Yao, Huan; Zhao, Hansen; Guo, Guangsheng; Zhang, Si-Chun; Zhang, Xinrong; Wang, Xiayan

doi:10.1039/d2sc02569h

Cited by 24 publications

(19 citation statements)

References 58 publications

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“…ILCEI-MS can simultaneously realize single-cell online dispersion and dilution-free MS injection with high sensitivity and high-throughput detection. To make the solvent compatible with MS detection while maintaining the osmotic pressure and physiological state of the cells to the maximum extent possible, the cells were resuspended in a 40 mmol/L ammonium formate aqueous solution based on previous studies . They were driven by a pneumatic-hydraulic injection device into a capillary with a 16-μm I.D.…”

Section: Resultsmentioning

confidence: 99%

“…One end of the nonetched capillary was inserted into a pressure-resistant small steel cylinder, and the other end of the gold-plated tip was installed at the front end of the mass spectrometer. The details of the device can be found in the previous work of our research group …”

Section: Methodsmentioning

confidence: 99%

“…The details of the device can be found in the previous work of our research group. 30 The ILCEI-MS consisted of an ILCEI ion source and an LTQ Orbitrap XL mass spectrometer (Thermo Scientific), separating around 6 mm. To minimize the detection error, the positive ion calibration solution was used for instrument correction before the measurement.…”

mentioning

confidence: 99%

“…This method is label-free and requires only simple sample preparation. The platform integrates high-sensitivity and high-throughput intact livingcell electro-launching ionization mass spectrometry (ILCEI-MS) 30 and metabolomics analysis. We applied this new platform to reveal the metabolic pathways significantly influenced by gefitinib.…”

mentioning

confidence: 99%

“…To make the solvent compatible with MS detection while maintaining the osmotic pressure and physiological state of the cells to the maximum extent possible, the cells were resuspended in a 40 mmol/L ammonium formate aqueous solution based on previous studies. 30 They were driven by a pneumatic-hydraulic injection device into a capillary with a 16-μm I.D. Whole cells were injected into the MS inlet from the narrow-bore constantinner-diameter emitter.…”

mentioning

confidence: 99%

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Single-Cell Metabolomics-Based Strategy for Studying the Mechanisms of Drug Action

et al. 2023

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Studying the mechanisms of drug antitumor activity at the single-cell level can provide information about the responses of cell subpopulations to drug therapy, which is essential for the accurate treatment of cancer. Due to the small size of single cells and the low contents of metabolites, metabolomics-based approaches to studying the mechanisms of drug action at the single-cell level are lacking. Herein, we develop a label-free platform for studying the mechanisms of drug action based on single-cell metabolomics (sMDA-scM) by integrating intact living-cell electro-launching ionization mass spectrometry (ILCEI-MS) with metabolomics analysis. Using this platform, we reveal that non-small-cell lung cancer (NSCLC) cells treated by gefitinib can be clustered into two cell subpopulations with different metabolic responses. The glutathione metabolic pathway of the subpopulation

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Single-Cell Metabolomics-Based Strategy for Studying the Mechanisms of Drug Action

et al. 2023

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Multi‐Mode/Signal Biosensors: Electrochemical Integrated Sensing Techniques

Han,

Wang,

Wang

2024

Adv Funct Materials

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Electrochemical (EC) analysis has emerged as a high‐sensitivity, reliable, cost‐effective, and rapidly evolving technique that has garnered significant attention across diverse domains. Furthermore, EC‐based techniques hold great potential for miniaturization and integration. The integration of EC techniques with diverse mode/signal (such as light, magnetic, and thermal signals, etc.) provides unique opportunities for biosensors to acquire more information through a single sensing platform. By coupling multiple signals or processing them logically, the detection accuracy can be further improved, and the probability of false positives or negatives can be minimized. In this review, a thorough analysis of multi‐ mode/signal sensors in the field of EC sensing is conducted, along with their integration with various sensing techniques (e.g., fluorescence, photothermal, colorimetry, microfluidic, etc.). The aim is to delve into the latest advances, potential applications, as well as challenges in multi‐mode/signal biosensors, where the utilization of multiple sensing modalities helps enhance accuracy, sensitivity, and selectivity. This review provides new insight into the synergistic effects of integrating EC sensing with other techniques, aiming to shed light on the near‐future developments in EC‐integrated multi‐mode/signal biosensors.

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High‐Throughput Single‐Cell, Single‐Mitochondrial DNA Assay Using Hydrogel Droplet Microfluidics

Park,

Kadam,

Atiyas

et al. 2024

Angewandte Chemie

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There is growing interest in understanding the biological implications of single cell heterogeneity and heteroplasmy of mitochondrial DNA (mtDNA), but current methodologies for single‐cell mtDNA analysis limit the scale of analysis to small cell populations. Although droplet microfluidics have increased the throughput of single‐cell genomic, RNA, and protein analysis, their application to sub‐cellular organelle analysis has remained a largely unsolved challenge. Here, we introduce an agarose‐based droplet microfluidic approach for single‐cell, single‐mtDNA analysis, which allows simultaneous processing of hundreds of individual mtDNA molecules within >10,000 individual cells. Our microfluidic chip encapsulates individual cells in agarose beads, designed to have a sufficiently dense hydrogel network to retain mtDNA after lysis and provide a robust scaffold for subsequent multi‐step processing and analysis. To mitigate the impact of the high viscosity of agarose required for mtDNA retention on the throughput of microfluidics, we developed a parallelized device, successfully achieving ~95% mtDNA retention from single cells within our microbeads at >700,000 drops/minute. To demonstrate utility, we analyzed specific regions of the single‐mtDNA using a multiplexed rolling circle amplification (RCA) assay. We demonstrated compatibility with both microscopy, for digital counting of individual RCA products, and flow cytometry for higher throughput analysis.

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Intact living-cell electrolaunching ionization mass spectrometry for single-cell metabolomics

Abstract: While single-cell mass spectrometry can reveal cellular heterogeneity and the molecular mechanisms of intracellular biochemical reactions, its application is limited by the insufficient detection sensitivity resulting from matrix interference and...

Cited by 24 publications

References 58 publications

Single-Cell Metabolomics-Based Strategy for Studying the Mechanisms of Drug Action

Single-Cell Metabolomics-Based Strategy for Studying the Mechanisms of Drug Action

Multi‐Mode/Signal Biosensors: Electrochemical Integrated Sensing Techniques

High‐Throughput Single‐Cell, Single‐Mitochondrial DNA Assay Using Hydrogel Droplet Microfluidics

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