SPEKcheck — fluorescence microscopy spectral visualisation and optimisation: a web application, javascript library, and data resource

Wellcome Open Research

article

Mick A. Phillips¹  David M. Susano Pinto¹  Ian M. Dobbie¹ 

[1] Micron Advanced Bioimaging Unit, Department of Biochemistry, University of Oxford

关键词: Microscopy;    Fluorescence;    Excitation;    Modelling;   

DOI  :  10.12688/wellcomeopenres.14703.1

学科分类：内科医学

来源: Wellcome

【 摘 要 】

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.

【 授权许可】

CC BY