Yapp Clarence, Rogers Catherine, Savitsky Pavel, Philpott Martin, Müller Susanne
Target Discovery Institute, NDMRB, University of Oxford, Oxford, OX3 7FZ, UK; Botnar Research Centre, NDORMS, University of Oxford, Oxford, OX3 7LD, UK.
Target Discovery Institute, NDMRB, University of Oxford, Oxford, OX3 7FZ, UK.
Biomed Opt Express. 2016 Jan 11;7(2):422-41. doi: 10.1364/BOE.7.000422. eCollection 2016 Feb 1.
Fluorescence Recovery After Photobleaching (FRAP) is an established method for validating chemical probes against the chromatin reading bromodomains, but so far requires constant human supervision. Here, we present Frapid, an automated open source code implementation of FRAP that fully handles cell identification through fuzzy logic analysis, drug dispensing with a custom-built fluid handler, image acquisition & analysis, and reporting. We successfully tested Frapid on 3 bromodomains as well as on spindlin1 (SPIN1), a methyl lysine binder, for the first time.
光漂白后荧光恢复(FRAP)是一种验证针对染色质读取溴结构域的化学探针的既定方法,但迄今为止需要持续的人工监督。在此,我们展示了Frapid,这是一种FRAP的自动化开源代码实现,它通过模糊逻辑分析、使用定制流体处理器进行药物分配、图像采集与分析以及报告,完全处理细胞识别。我们首次在3个溴结构域以及甲基赖氨酸结合蛋白纺锤体相关蛋白1(SPIN1)上成功测试了Frapid。