Single-cell capillary electrophoresis mass spectrometry (CE-MS) is a promising platform to analyze cellular contents and probe cell heterogeneity. However, current single-cell CE-MS methods often rely on offline microsampling processes and may demonstrate low sampling precision and accuracy. We have recently developed an electrospray-assisted device, , for low-volume sample extraction. With the spray-capillary, low-volume samples (pL-nL) are drawn into the sampling end of the device, which can be used directly for CE separation and online MS detection. Here, we redesigned the spray-capillary by utilizing a capillary with a <15 μm tapered tip so that it can be directly inserted into single cells for sample collection and on-capillary CE-MS analysis. We evaluated the performance of the modified spray-capillary by performing single-cell microsampling on single onion cells with varying sample injection times and direct MS analysis or online CE-MS analysis. We have demonstrated, for the first time, online sample collection and CE-MS for the analysis of single cells. This application of the modified spray-capillary device facilitates the characterization and relative quantification of hundreds of metabolites in single cells.
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http://dx.doi.org/10.1021/acs.analchem.0c04624 | DOI Listing |
Anal Chem
December 2024
Department of Chemistry and Chemical Biology, McMaster University, Hamilton, Ontario L8S 4M1, Canada.
Mass spectrometry (MS)-based metabolomics often rely on separation techniques when analyzing complex biological specimens to improve method resolution, metabolome coverage, quantitative performance, and/or unknown identification. However, low sample throughput and complicated data preprocessing procedures remain major barriers to affordable metabolomic studies that are scalable to large populations. Herein, we introduce PeakMeister as a new software tool in the R statistical environment to enable standardized processing of serum metabolomic data acquired by multisegment injection-capillary electrophoresis-mass spectrometry (MSI-CE-MS), a high-throughput separation platform (<4 min/sample) which takes advantage of a serial injection format of 13 samples within a single analytical run.
View Article and Find Full Text PDFMetabolites
November 2024
Department of Intensive Care Medicine, Institute of Science Tokyo, 1-5-45 Yushima, Bunkyo City 113-8510, Japan.
A dysregulated metabolism has been studied as a key aspect of the COVID-19 pathophysiology, but its longitudinal progression in severe cases remains unclear. In this study, we aimed to investigate metabolic dysregulation over time in patients with severe COVID-19 requiring mechanical ventilation (MV). In this single-center, prospective, observational study, we obtained 236 serum samples from 118 adult patients on MV in an ICU.
View Article and Find Full Text PDFGels
December 2024
Horváth Csaba Memorial Laboratory of Bioseparation Sciences, Research Center for Molecular Medicine, Faculty of Medicine, Doctoral School of Medicine, University of Debrecen, 4032 Debrecen, Hungary.
Hydrogels like agarose have long been used as sieving media for the electrophoresis-based analysis of biopolymers. During gelation, the individual agarose strands tend to form hydrogen-bond mediated double-helical structures, allowing thermal reversibility and adjustable pore sizes for molecular sieving applications. The addition of tetrahydroxyborate to the agarose matrix results in transitional chemical cross-linking, offering an additional pore size adjusting option.
View Article and Find Full Text PDFFront Genome Ed
December 2024
Agronomy Department, Plant Molecular and Cellular Biology Program, Genetics Institute, University of Florida, IFAS-Institute of Food and Agricultural Science, Gainesville, FL, United States.
Sugarcane ( spp.) is an important biofuel feedstock and a leading source of global table sugar. hybrid cultivars are highly polyploid (2n = 100-130), containing large numbers of functionally redundant hom(e)ologs in their genomes.
View Article and Find Full Text PDFBiotechniques
December 2024
Department of Biology, Montclair State University, Montclair, NJ, USA.
Microsatellites are present in mitochondria, chloroplast, and nuclear DNA, but nuclear microsatellites are more useful genetic tools than those in plastids or mitochondria. Plastid and mitochondrial microsatellites have been identified in the model plant (liverwort), but no laboratory has published information on nuclear microsatellite loci. The aim of this study was to detect novel nuclear markers in the most commonly employed liverwort species, design PCR primers that would allow amplification, and characterize the subsequently generated loci.
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