Affiliation:
1. Department of Biomedical Engineering National University of Singapore Singapore 117583 Singapore
2. Singapore Immunology Network Agency for Science Technology and Research Singapore 138648 Singapore
3. Institute for Health Innovation & Technology National University of Singapore Singapore 117599 Singapore
4. Tissue Engineering Programme National University of Singapore Singapore 117510 Singapore
Abstract
AbstractLongitudinal analysis of intracellular contents including gene and protein expression is crucial for deciphering the fundamentally dynamic nature of cells. This offers invaluable insights into complex tissue composition and behavior, and drives progress in disease diagnosis, biomarker discovery, and drug development. Traditional longitudinal analysis workflows, involving the destruction of cells at various timepoints, limit insights to singular moments and fail to account for cellular heterogeneity. Current non‐destructive approaches, like temporal modeling with single‐cell ribonucleic acid sequencing (RNA‐seq) and live‐cell fluorescence imaging, either rely on biological assumptions or possess the risk of cellular perturbation. Recent advances in nanoscale technologies for non‐destructive intracellular content extraction offer a promising solution to these challenges. These novel methods work at the nanoscale to non‐destructively access cellular membranes and can be broadly classified into three mechanisms: tip‐facilitated aspiration, membrane‐based, and probe‐based methods. This perspective focuses on these emerging nanotechnologies for repeated intracellular content extraction. Their potential in longitudinal analysis is discussed, the critical requirements for effective repeated sampling are addressed, and the suitability of each technique for various applications is explored. Furthermore, unresolved challenges in repeated sampling are highlighted to encourage further research in this growing field.
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2 articles.
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