小柯机器人

美国德克萨斯大学奥斯汀分校Adela Ben-Yakar研究组发现，飞秒激光显微切割技术可用于分离再生的线虫神经元来进行单细胞RNA测序。该项研究成果于2023年3月16日在线发表在《自然—方法学》杂志上。

研究人员提出了飞秒激光显微切割（fs-LM），这是一种单细胞分离方法，通过利用飞秒激光消融的微米级精度，直接从活体组织中分离出特定细胞。结果表明，fs-LM有利于通过单细胞RNA测序（scRNA-seq）进行敏感和特异的基因表达分析，同时减轻了组织分离引起的与压力有关的转录伪影。研究人员对fs-LM分离出的再生神经元进行scRNA-seq，发现转录程序分别与野生型和dlk-1(0)动物的再生成功或失败相关。这种方法还能够研究同一类型的神经元所显示的异质性，并发现与轴突再生率相关的基因模块的表达模式。这些结果建立了fs-LM作为一种空间分辨率的单细胞分离方法，可用于表型到基因型的映射。

研究人员表示，人们对神经再生的理解可以通过在模型生物（例如线虫）中以单神经元的分辨率描绘其基本的分子活动来加强。现有的细胞分离技术无法从线虫中分离出具有特定再生表型的神经元。

附：英文原文

Title: Femtosecond laser microdissection for isolation of regenerating C. elegans neurons for single-cell RNA sequencing

Author: Zhao, Peisen, Mondal, Sudip, Martin, Chris, DuPlissis, Andrew, Chizari, Shahab, Ma, Ke-Yue, Maiya, Rajani, Messing, Robert O., Jiang, Ning, Ben-Yakar, Adela

Issue&Volume: 2023-03-16

Abstract: Our understanding of nerve regeneration can be enhanced by delineating its underlying molecular activities at single-neuron resolution in model organisms such as Caenorhabditis elegans. Existing cell isolation techniques cannot isolate neurons with specific regeneration phenotypes from C. elegans. We present femtosecond laser microdissection (fs-LM), a single-cell isolation method that dissects specific cells directly from living tissue by leveraging the micrometer-scale precision of fs-laser ablation. We show that fs-LM facilitates sensitive and specific gene expression profiling by single-cell RNA sequencing (scRNA-seq), while mitigating the stress-related transcriptional artifacts induced by tissue dissociation. scRNA-seq of fs-LM isolated regenerating neurons revealed transcriptional programs that are correlated with either successful or failed regeneration in wild-type and dlk-1 (0) animals, respectively. This method also allowed studying heterogeneity displayed by the same type of neuron and found gene modules with expression patterns correlated with axon regrowth rate. Our results establish fs-LM as a spatially resolved single-cell isolation method for phenotype-to-genotype mapping.

DOI: 10.1038/s41592-023-01804-3

Source: https://www.nature.com/articles/s41592-023-01804-3

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex