Phillips Mick A, Susano Pinto David M, Dobbie Ian M
Micron Advanced Bioimaging Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX13QU, UK.
Wellcome Open Res. 2018 Jul 30;3:92. doi: 10.12688/wellcomeopenres.14703.1. eCollection 2018.
Advanced fluorescence imaging methods require careful matching of excitation sources, dichroics, emission filters, detectors, and dyes to operate at their best. This complex task is often left to guesswork, preventing optimal dye:filter combinations, particularly for multicolour applications. To overcome this challenge we developed SPEKcheck, a web application to visualise the efficiency of the light path in a fluorescence microscope. The software reports values for the excitation efficiency of a dye, the collection efficiency of the emitted fluorescence, and a "brightness" score, allowing easy comparison between different fluorescent labels. It also displays a spectral plot of various elements in the configuration, enabling users to readily spot potential problems such as low efficiency excitation, emission, or high bleedthrough. It serves as an aid to exploring the performance of different dyes and filter sets.
先进的荧光成像方法需要仔细匹配激发源、二向色镜、发射滤光片、探测器和染料，以实现最佳运行。这项复杂的任务通常只能靠猜测，这就无法实现最佳的染料：滤光片组合，尤其是在多色应用中。为了克服这一挑战，我们开发了SPEKcheck，这是一个用于可视化荧光显微镜光路效率的网络应用程序。该软件会报告染料的激发效率、发射荧光的收集效率以及一个“亮度”分数的值，便于对不同的荧光标记进行比较。它还会显示配置中各种元件的光谱图，使用户能够轻松发现潜在问题，如激发效率低、发射效率低或高串扰。它有助于探索不同染料和滤光片组的性能。
