An open source 16-channel fluidics system for automating sequential fluorescent in situ hybridization (FISH)-based imaging.
An open source 16-channel fluidics system for automating sequential fluorescent in situ hybridization (FISH)-based imaging.
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DOI：
10.1016/j.ohx.2022.e00343
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发表时间：
2022-10
期刊：
影响因子：
2.2
通讯作者：
Beliveau, Brian J.
中科院分区：
文献类型：
--
作者：
Deng, Zhaojie;Beliveau, Brian J.
荧光原位杂交(FISH)可以提供固定细胞和组织中DNA/RNA靶标的空间信息。然而，由于液体处理的需求，随着轮次的增加，使用连续轮次杂交的多路FISH成像的工作流程很快变得繁琐。在这里，我们提出了一个开源和低成本的流体系统，它是专门为自动化顺序鱼类成像的工作流程而构建的。我们的系统具有一个带有16个可寻址通道的流体模块，其中的流量是正压驱动的，并由电磁阀开启/关闭，以便将FISH试剂传输到样品中。我们的系统还包括一个带有主印刷电路板的控制器，可以控制多达120个电磁阀，并允许用户通过串口通信控制流体模块。我们通过14轮连续杂交和成像，以HeLa细胞中的α卫星重复序列为靶点，展示了该系统自动和稳健的液体交换。我们预计，这个简单灵活的系统将对研究人员在一系列实验系统中进行多路原位分析有用。
Fluorescent in situ hybridization (FISH) can provide spatial information about DNA/RNA targets in fixed cells and tissues. However, the workflows of multiplexed FISH-based imaging that use sequential rounds of hybridization quickly become laborious as the number of rounds increases because of liquid handling demands. Here, we present an open-source and low-cost fluidics system that is purpose built for automating the workflows of sequential FISH-based imaging. Our system features a fluidics module with 16 addressable channels in which flow is positive pressure-driven and switched on/off by solenoid valves in order to transfer FISH reagents to the sample. Our system also includes a controller with a main printed circuit board that can control up to 120 solenoid valves and allows users to control the fluidics module via serial communication. We demonstrate the automatic and robust fluid exchange with this system by targeting the alpha satellite repeat in HeLa cell with 14 rounds of sequential hybridization and imaging. We anticipate that this simple and flexible system will be of utility to researchers performing multiplexed in situ assays in a range of experimental systems.
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影响因子：
48
作者：
Lubeck, Eric;Cai, Long
通讯作者：
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DOI：
10.1126/science.aau1783
发表时间：
2018-10-26
期刊：
Science (New York, N.Y.)
影响因子：
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作者：
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通讯作者：
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64.5
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