Hyde James, MacNicol Melanie, Odle Angela, Garcia-Rill Edgar
Center for Translational Neuroscience, Department of Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences, Little Rock, AR, USA.
Center for Translational Neuroscience, Department of Neurobiology & Developmental Sciences, University of Arkansas for Medical Sciences, Little Rock, AR, USA.
J Neurosci Methods. 2014 Dec 30;238:82-7. doi: 10.1016/j.jneumeth.2014.09.012. Epub 2014 Sep 22.
In recent years, 3D printing technology has become inexpensive and simple enough that any lab can own and use one of these printers.
We explored the potential use of 3D printers for quickly and easily producing in vitro slice chambers for patch clamp electrophysiology. Slice chambers were produced using five available plastics: ABS, PLA, Nylon 618, Nylon 680, and T-glase. These "lab-made" chambers were also made using stereolithography through a professional printing service (Shapeways). This study measured intrinsic membrane properties of neurons in the brain stem pedunculopontine nucleus (PPN) and layer V pyramidal neurons in retrosplenial cortex.
Nylon 680 and T-glase significantly hyperpolarized PPN neurons. ABS increased input resistance, decreased action potential amplitude, and increased firing frequency in pyramidal cortical neurons. To test long term exposure to each plastic, human neuroblastoma SHSY5Y cell cultures were exposed to each plastic for 1 week. ABS decreased cell counts while Nylon 618 and Shapeways plastics eliminated cells. Primary mouse pituitary cultures were also tested for 24-h exposure. ABS decreased cell counts while Nylon 618 and Shapeways plastics dramatically decreased cell counts.
Chambers can be quickly and inexpensively printed in the lab. ABS, PLA, Nylon 680, and T-glase plastics would suffice for many experiments instead of commercially produced slice chambers.
While these technologies are still in their infancy, they represent a powerful addition to the lab environment. With careful selection of print material, slice chambers can be quickly and inexpensively manufactured in the lab.
近年来，3D打印技术已变得足够便宜且操作简单，任何实验室都可以拥有并使用其中一台打印机。
我们探索了3D打印机在快速轻松生产用于膜片钳电生理的体外切片槽方面的潜在用途。使用五种可用塑料制作切片槽：丙烯腈-丁二烯-苯乙烯共聚物（ABS）、聚乳酸（PLA）、尼龙618、尼龙680和T-玻璃。这些“实验室自制”的切片槽也通过专业打印服务（Shapeways）使用立体光刻法制作。本研究测量了脑干脚桥核（PPN）中的神经元以及压后皮质第V层锥体神经元的内在膜特性。
尼龙680和T-玻璃使PPN神经元显著超极化。ABS增加了锥体皮质神经元的输入电阻，降低了动作电位幅度，并增加了放电频率。为了测试长期暴露于每种塑料的影响，将人神经母细胞瘤SHSY5Y细胞培养物暴露于每种塑料1周。ABS减少了细胞数量，而尼龙618和Shapeways的塑料使细胞消失。原代小鼠垂体培养物也进行了24小时暴露测试。ABS减少了细胞数量，而尼龙618和Shapeways的塑料显著减少了细胞数量。
切片槽可以在实验室中快速且廉价地打印出来。ABS、PLA、尼龙680和T-玻璃塑料足以满足许多实验，而无需使用商业生产的切片槽。
虽然这些技术仍处于起步阶段，但它们为实验室环境增添了强大助力。通过仔细选择打印材料，可以在实验室中快速且廉价地制造切片槽。
