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Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2026,4,21]]},"abstract":"<jats:p>Monitoring molecular activities within single live cells is vital for understanding cellular differentiation, senescence, heterogeneity, and disease progression. However, conventional single-cell analyses often rely on micromanipulation or extraction followed by downstream measurements, which cannot capture in situ real-time dynamics. Fluorescent labeling and electrochemical methods provide temporal resolution but face limitations in labeling, substrate scope, and multiplexing. Here, we present a nanopore probe that enables real-time, multiplexed monitoring of intracellular activities in single live cells. The device integrates an aluminum oxide nanostraw membrane for molecular extraction and a glass nanopore membrane for single-channel electrical detection. Using a hippocampal neuron model of ischemia-hypoxia, we simultaneously tracked dynamic changes in intracellular glutamate, ascorbic acid, and adenosine triphosphate\u2014three key molecules involved in oxygen-glucose deprivation-induced neuronal edema. Our findings establish this nanopore probe as a powerful platform for real-time, label-free molecular profiling at the single-cell level, opening opportunities for studying disease mechanisms and therapeutic responses.<\/jats:p>","DOI":"10.1073\/pnas.2529161123","type":"journal-article","created":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T17:36:48Z","timestamp":1776274608000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":2,"title":["In situ and real-time monitoring of intracellular activities in single live cells using a nanopore probe"],"prefix":"10.1073","volume":"123","author":[{"given":"Xiaobin","family":"Huang","sequence":"first","affiliation":[{"name":"Beijing National Laboratory for Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences","place":["Beijing, China"]},{"name":"School of Chemical Sciences, University of Chinese Academy of 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