Xu Xiaolong, Jia Jianbo, Guo Mingquan
School of Biotechnology and Health Science, Wuyi University, Jiangmen, China.
CAS Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Chinese Academy of Sciences, Wuhan, China.
Front Chem. 2020 Aug 20;8:718. doi: 10.3389/fchem.2020.00718. eCollection 2020.
The research in endogenous biomolecules from a single cell has grown rapidly in recent years since it is critical for dissecting and scrutinizing the complexity of heterogeneous tissues, especially under pathological conditions, and it is also of key importance to understand the biological processes and cellular responses to various perturbations without the limitation of population averaging. Although conventional techniques, such as micromanipulation or cell sorting methods, are already used along with subsequent molecular examinations, it remains a big challenge to develop new approaches to manipulate and directly extract small quantities of cytosol from single living cells. In this sense, nanostructure or nanomaterial may play a critical role in overcoming these challenges in cellular manipulation and extraction of very small quantities of cells, and provide a powerful alternative to conventional techniques. Since the nanostructures or nanomaterial could build channels between intracellular and extracellular components across cell membrane, through which cytosol could be pumped out and transferred to downstream analyses. In this review, we will first brief the traditional methods for single cell analyses, and then shift our focus to some most promising methods for single-cell sampling with nanostructures, such as glass nanopipette, nanostraw, carbon nanotube probes and other nanomaterial. In this context, particular attentions will be paid to their principles, preparations, operations, superiorities and drawbacks, and meanwhile the great potential of nano-materials for single-cell sampling will also be highlighted and prospected.
近年来,对单细胞内源性生物分子的研究发展迅速,因为这对于剖析和审视异质组织的复杂性至关重要,尤其是在病理条件下,而且在不受群体平均限制的情况下理解生物过程和细胞对各种扰动的反应也至关重要。尽管传统技术,如显微操作或细胞分选方法,已与后续分子检测一起使用,但开发新方法以操纵并直接从单个活细胞中提取少量胞质溶胶仍然是一项巨大挑战。从这个意义上说,纳米结构或纳米材料可能在克服细胞操作和极少量细胞提取方面的这些挑战中发挥关键作用,并为传统技术提供有力替代方案。因为纳米结构或纳米材料可以在跨细胞膜的细胞内和细胞外成分之间构建通道,通过该通道可以将胞质溶胶泵出并转移到下游分析中。在本综述中,我们将首先简要介绍单细胞分析的传统方法,然后将重点转向一些最有前景的利用纳米结构进行单细胞采样的方法,如玻璃纳米吸管、纳米吸管、碳纳米管探针和其他纳米材料。在此背景下,将特别关注它们的原理、制备、操作、优点和缺点,同时也将突出并展望纳米材料在单细胞采样方面的巨大潜力。