A microfluidic device for TEM sample preparation,Lab on a Chip

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A microfluidic device for TEM sample preparation
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-10-09 , DOI: 10.1039/d0lc00724b
Janosch Hauser _{1,

2,

3,

4} , Gustaf Kylberg _{4,

5,

6} , Mathieu Colomb-Delsuc _{4,

5,

6} , Göran Stemme _{1,

2,

3,

4} , Ida-Maria Sintorn _{4,

5,

6} , Niclas Roxhed _{1,

2,

3,

4}

Affiliation

Transmission electron microscopy (TEM) allows for visualizing and analyzing viral particles and has become a vital tool for the development of vaccines and biopharmaceuticals. However, appropriate TEM sample preparation is typically done manually which introduces operator-based dependencies and can lead to unreliable results. Here, we present a capillary-driven microfluidic single-use device that prepares a TEM grid with minimal and non-critical user interaction. The user only initiates the sample preparation process, waits for about one minute and then collects the TEM grid, ready for imaging. Using Adeno-associated virus (AAV) particles as the sample and NanoVan® as the stain, we demonstrate microfluidic consistency and show that the sample preparation quality is sufficient for automated image analysis. We further demonstrate the versatility of the microfluidic device by preparing two protein complexes for TEM investigations using two different stain types. The presented TEM sample preparation concept could alleviate the problems associated with human inconsistency in manual preparation protocols and allow for non-specialists to prepare TEM samples.

中文翻译：

用于TEM样品制备的微流控设备

透射电子显微镜（TEM）可以可视化和分析病毒颗粒，并已成为开发疫苗和生物制药的重要工具。但是，适当的TEM样品制备通常是手动完成的，这会引入基于操作员的依赖性，并且可能导致结果不可靠。在这里，我们介绍了一种毛细管驱动的微流控单次使用设备，该设备可通过最少且非关键的用户交互来准备TEM网格。用户仅启动样品制备过程，等待大约一分钟，然后收集TEM网格，准备进行成像。使用腺相关病毒（AAV）颗粒作为样品，使用NanoVan®作为染色剂，我们证明了微流体的一致性，并表明样品制备质量足以用于自动化图像分析。我们通过使用两种不同的染色剂类型准备用于TEM研究的两种蛋白质复合物，进一步证明了微流控设备的多功能性。提出的TEM样品制备概念可以缓解与人工制备方案中的人为不一致有关的问题，并允许非专业人员制备TEM样品。

更新日期：2020-11-03

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