Pan Peng, Qin Zhen, Sun William, Zhou Yuxiao, Wang Shaojia, Song Pengfei, Wang Yong, Ru Changhai, Wang Xin, Calarco John, Liu Xinyu
Department of Mechanical and Industrial Engineering, University of Toronto, 5 King's College Road, Toronto, Ontario M5S 3G8 Canada.
Upper Canada College, 200 Lonsdale Road, Toronto, Ontario M4V 1W6 Canada.
Microsyst Nanoeng. 2023 Feb 21;9:17. doi: 10.1038/s41378-023-00485-4. eCollection 2023.
embryos have been widely used to study cellular processes and developmental regulation at early stages. However, most existing microfluidic devices focus on the studies of larval or adult worms rather than embryos. To accurately study the real-time dynamics of embryonic development under different conditions, many technical barriers must be overcome; these can include single-embryo sorting and immobilization, precise control of the experimental environment, and long-term live imaging of embryos. This paper reports a spiral microfluidic device for effective sorting, trapping, and long-term live imaging of single embryos under precisely controlled experimental conditions. The device successfully sorts embryos from a mixed population of at different developmental stages via Dean vortices generated inside a spiral microchannel and traps the sorted embryos at single-cell resolution through hydrodynamic traps on the sidewall of the spiral channel for long-term imaging. Through the well-controlled microenvironment inside the microfluidic device, the response of the trapped embryos to mechanical and chemical stimulation can be quantitatively measured. The experimental results show that a gentle hydrodynamic force would induce faster growth of embryos, and embryos developmentally arrested in the high-salinity solution could be rescued by the M9 buffer. The microfluidic device provides new avenues for easy, rapid, high-content screening of embryos.
胚胎已被广泛用于研究早期阶段的细胞过程和发育调控。然而,大多数现有的微流控装置专注于幼虫或成虫的研究,而非胚胎。为了准确研究不同条件下胚胎发育的实时动态,必须克服许多技术障碍;这些障碍包括单胚胎分选和固定、实验环境的精确控制以及胚胎的长期活体成像。本文报道了一种螺旋微流控装置,用于在精确控制的实验条件下对单个胚胎进行有效分选、捕获和长期活体成像。该装置通过螺旋微通道内产生的迪恩涡旋成功地从不同发育阶段的混合群体中对胚胎进行分选,并通过螺旋通道侧壁上的流体动力阱以单细胞分辨率捕获分选后的胚胎,以便进行长期成像。通过微流控装置内良好控制的微环境,可以定量测量捕获的胚胎对机械和化学刺激的反应。实验结果表明,温和的流体动力会诱导胚胎更快生长,并且在高盐溶液中发育停滞的胚胎可以通过M9缓冲液挽救。该微流控装置为胚胎的简便、快速、高内涵筛选提供了新途径。