用于双电极电压钳电生理学的经济实惠的三维 (3D) 打印记录室。

Ian Sk Shogren, Jean P Gonzales, Linda M Boland
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摘要

爪蟾卵母细胞双电极电压钳(TEVC)电生理学是研究膜转运蛋白生理学和药理学的常用方法。本科生可以在神经科学或生理学课程和/或教师指导的研究经历中学习使用 TEVC 方法。该方法面临的挑战包括市售记录室的成本(尤其是当实验室需要多份记录室时),以及使用琼脂桥、稳定溶液流动和尽量减少溶液吸入噪音所需的额外时间和专业知识。如果能提供一种克服这些困难的低成本、易使用的记录室,就能降低本科生成功的门槛,同时还能支持出版质量的记录。为了解决这些问题,我们利用立体光刻技术(一种三维打印工艺)开发了一种记录室。生理学(PhISio)记录室有两种溶液吸入方式可供选择,以满足个人偏好;优化了预制琼脂桥的放置,以实现层流并减少启动日常实验的时间延迟;最大限度地减少了灌注过程中改变溶液高度和吸入噪音所带来的挑战。我们比较了 PhISio 试验室与市场上销售的华纳仪器公司 RC-1Z 试验室在对爪蟾卵母细胞表达的内向整流钾通道进行电生理记录时的功能。在随时间变化的溶液方面,PhISio 试验室与 RC-1Z 试验室产生了相同的结果,而且对于新手和有经验的电生理学家来说,PhISio 试验室都具有一些操作优势,是市售 TEVC 记录室的经济、方便的替代品。
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An affordable Three-Dimensional (3D) Printed Recording Chamber for Two-Electrode Voltage Clamp Electrophysiology.

Two electrode voltage-clamp (TEVC) electrophysiology in Xenopus oocytes is a common approach to studying the physiology and pharmacology of membrane transport proteins. Undergraduates may learn to use TEVC methodology in neuroscience or physiology courses and/or in faculty-mentored research experiences. Challenges with the methodology include the cost of commercially available recording chambers, especially when a lab needs multiple copies, and the additional time and expertise needed to use agar bridges and to stabilize solution flow and minimize noise from solution aspiration. Offering a low-cost and accessible recording chamber that overcomes these challenges would lower the barriers to success for undergraduates while also supporting publication-quality recordings. To address these issues, we developed a recording chamber using stereolithography, a 3D printing process. The physiology (PhISio) recording chamber features two options for solution aspiration that allow for individual preferences, optimizes placement of pre-made agar bridges to achieve laminar flow and reduce the time delays in initiating daily experiments, and minimizes the challenges of changing solution height and aspiration noise during perfusion. We compared the functionality of the PhISio chamber with a commercially available Warner Instruments RC-1Z chamber in electrophysiological recordings of inwardly rectifying potassium channels expressed in Xenopus oocytes. The PhISio chamber produced equivalent results to the RC-1Z chamber with respect to time-dependent solution changes and has several operational advantages for both new and experienced electrophysiologists, providing an affordable and convenient alternative to commercially available TEVC recording chambers.

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