Fluorescence Spectrum Viewer

Using this applet

Zooming in/out

To Zoom in: Click and drag from left to right, then release to zoom the selected area. To return to the original view, double click on the canvas.

Assessing spillover estimates

For any given filter/fluorophore combination, click and hold over a filter area to view normalized spillover estimates (where data permits) and the filter efficiency for the primary fluorochrome. Relevant caveats: The representative fluorometer data loaded into the applet are from a variety of sources, and the curves are not necessarily reflective of all reagent conjugates, especially fluorescent tandems like PE-Cy7 since PMT quantum efficiency falls off towards the far red. Additionally, keep in mind that not all spectra were obtained from the same fluorometer (see below). There are three methods to estimate spillover:

% Total

The spill estimate is the percent of the total (measured + unmeasured).

% Measured

The spill estimate is the percent of measured (varies with primary filter).

Average %

The average percent within the spill filter.

Important Information

The spectra presented here have been collected on a fluorometer, and are not necessarily corrected for the photomultiplier tube (PMT) that might be in your instrument. The spectral sensitivity of the fluorometer and PMT used has an effect on the measured relative "intensities" of the yellow and near infra red fluorescence. A typical fluorometer has a "good" red sensitive PMT, but can often be more sensitive (~3X depending on fluorometer) in the yellow than in the near infra red. This is especially relevant to evaluating tandems such as PE-Cy7, where the PE spillover component is typically not as strong as the representative curves might suggest.