Shogren Ian Sk, Gonzales Jean P, Boland Linda M
Biology Department, University of Richmond, Richmond, VA 23713.
J Undergrad Neurosci Educ. 2023 May 19;21(2):A91-A96. doi: 10.59390/VSKD5667. eCollection 2023 Spring.
Two electrode voltage-clamp (TEVC) electrophysiology in 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 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.
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。本科生可能会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个副本时,以及使用琼脂桥、稳定溶液流动和最小化溶液抽吸产生的噪音所需的额外时间和专业知识。提供一种低成本且易于使用的记录室来克服这些挑战,将降低本科生取得成功的障碍,同时也支持高质量的发表记录。为了解决这些问题,我们使用立体光刻技术(一种3D打印工艺)开发了一种记录室。生理学(PhISio)记录室具有两种溶液抽吸选项,可满足个人偏好,优化预制琼脂桥的放置以实现层流并减少每日实验开始时的时间延迟,并最大限度地减少灌注过程中溶液高度变化和抽吸噪音的挑战。我们在卵母细胞中表达的内向整流钾通道的电生理记录中,将PhISio记录室的功能与市售的华纳仪器RC-1Z记录室进行了比较。在随时间变化的溶液变化方面,PhISio记录室产生的结果与RC-1Z记录室相当,并且对新老电生理学家都有几个操作优势,为市售的TEVC记录室提供了一种经济实惠且方便的替代方案。