Simultaneous spectral illumination of microplates for high-throughput optogenetics and photobiology.

IF 2.9 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Arend Vogt, Raik Paulat, Daniel Parthier, Verena Just, Michal Szczepek, Patrick Scheerer, Qianzhao Xu, Andreas Möglich, Dietmar Schmitz, Benjamin R Rost, Nikolaus Wenger
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引用次数: 0

Abstract

The biophysical characterization and engineering of optogenetic tools and photobiological systems has been hampered by the lack of efficient methods for spectral illumination of microplates for high-throughput analysis of action spectra. Current methods to determine action spectra only allow the sequential spectral illumination of individual wells. Here we present the open-source RainbowCap-system, which combines LEDs and optical filters in a standard 96-well microplate format for simultaneous and spectrally defined illumination. The RainbowCap provides equal photon flux for each wavelength, with the output of the LEDs narrowed by optical bandpass filters. We validated the RainbowCap for photoactivatable G protein-coupled receptors (opto-GPCRs) and enzymes for the control of intracellular downstream signaling. The simultaneous, spectrally defined illumination provides minimal interruption during time-series measurements, while resolving 10 nm differences in the action spectra of optogenetic proteins under identical experimental conditions. The RainbowCap is also suitable for studying the spectral dependence of light-regulated gene expression in bacteria, which requires illumination over several hours. In summary, the RainbowCap provides high-throughput spectral illumination of microplates, while its modular, customizable design allows easy adaptation to a wide range of optogenetic and photobiological applications.

用于高通量光遗传学和光生物学的微孔板同步光谱照明。
由于缺乏对微孔板进行光谱照射以进行高通量作用光谱分析的有效方法,光遗传工具和光生物系统的生物物理特征描述和工程设计受到了阻碍。目前确定作用光谱的方法只能对单个孔进行顺序光谱照明。在此,我们介绍开源的 RainbowCap 系统,该系统在标准 96 孔微孔板格式中结合了 LED 和光学滤光片,可同时进行光谱照明。RainbowCap 为每个波长提供相同的光通量,LED 的输出通过光学带通滤光片缩小。我们对 RainbowCap 进行了验证,可用于光激活 G 蛋白偶联受体(opto-GPCR)和酶,以控制细胞内的下游信号传导。在进行时间序列测量时,光谱定义的同步照明可提供最小的中断,同时在相同的实验条件下,可分辨光遗传蛋白作用光谱中 10 nm 的差异。RainbowCap 还适用于研究细菌中光调控基因表达的光谱依赖性,这需要几个小时的照明。总之,RainbowCap 可为微孔板提供高通量光谱照明,其模块化、可定制的设计可轻松适应各种光遗传学和光生物学应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biological Chemistry
Biological Chemistry 生物-生化与分子生物学
CiteScore
7.20
自引率
0.00%
发文量
63
审稿时长
4-8 weeks
期刊介绍: Biological Chemistry keeps you up-to-date with all new developments in the molecular life sciences. In addition to original research reports, authoritative reviews written by leading researchers in the field keep you informed about the latest advances in the molecular life sciences. Rapid, yet rigorous reviewing ensures fast access to recent research results of exceptional significance in the biological sciences. Papers are published in a "Just Accepted" format within approx.72 hours of acceptance.
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