Sweeney Kieran, Moreno Morales Neydis, Burmeister Zachary, Nimunkar Amit J, McClean Megan N
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, United States.
MethodsX. 2019 Jun 13;6:1480-1488. doi: 10.1016/j.mex.2019.06.008. eCollection 2019.
Optogenetic systems use genetically-encoded light-sensitive proteins to control and study cellular processes. As the number and quality of these systems grows, there is an increasing need for user-friendly and flexible hardware to provide programmed illumination to cultures of cells. One platform which satisfies this need for a variety of optogenetic systems and organisms is the Light Plate Apparatus (LPA), which delivers a controlled light dose to each well of a 24-well plate. Experimental reproducibility requires appropriate calibration to produce accurate light doses within individual wells of the LPA and between LPAs. In this study, we present an easy and accurate method for calibrating the LPA. In particular, we: •developed a 3D printed adaptor and MATLAB code to allow rapid measurement of irradiance produced by the LPA and subsequent calibration•provide appropriate code and methodology for generating a standard curve for each LPA•demonstrate the utility and accuracy of this method between users and LPAs.
光遗传学系统利用基因编码的光敏蛋白来控制和研究细胞过程。随着这些系统的数量和质量不断增加,对用户友好且灵活的硬件的需求也日益增长,以便为细胞培养物提供程序化照明。满足各种光遗传学系统和生物体这一需求的一个平台是光板装置(LPA),它能向24孔板的每个孔提供可控的光剂量。实验的可重复性需要进行适当校准,以便在LPA的各个孔内以及不同LPA之间产生准确的光剂量。在本研究中,我们提出了一种简单而准确的LPA校准方法。具体而言,我们:•开发了一个3D打印适配器和MATLAB代码,以便快速测量LPA产生的辐照度并进行后续校准•提供了为每个LPA生成标准曲线的适当代码和方法•展示了该方法在不同用户和LPA之间的实用性和准确性。