Microscope Pages
When using an online spectra viewer to decide which fluorophores might work best for their experiment, researchers frequently load a set of fluorescence filter spectra that match the filters on their instrument, and then compare those with a set of fluorophore spectra. Microscope pages are specifically designed to simplify this process by allowing users to save the components of their microscope at FPbase so that they do not need to re-enter filters every time they use the viewer. The result is a custom, microscope-specific spectra viewer with a dedicated URL that can be shared or embedded elsewhere (such as a lab or core facility website). Furthermore, “optical configurations” (combinations of excitation, dichroic, and emission filters) can be specified, making it easy to switch between the various filter arrangements that a user might have on their microscope. With the addition of light source(s) and camera spectra, relatively complete calculations can be made for the excitation and collection efficiency of a given optical configuration/fluorophore combination. Additionally, we find the visual representation of these efficiencies provided by the spectra-viewer to be useful in teaching scenarios. I specifically hope to see multi-user facilities (such as core facilities) benefit from this tool and welcome suggestions for improvement or requests for help in setting up a microscope page. Future embellishments may include cross-database calculations of fluorophore efficiency and dye-pair bleed-through that may help to assist users in choosing a set of probes well matched to their hardware. Note however, FP maturation and relative expression level are primary determinants of apparent brightness and bleedthrough, and neither can be predicted in any spectra-viewer!
Last modified 4yr ago