Guide For Using the Fluorescence SpectraViewer

(B) Adjusting spectra overlap—in the Spillover Table, the gray boxes are the percentage emission capture from targeted fluorophore using the specific laser and the emission filter or channel.

Outside of the gray boxes are the spillover or spectra overlap values. In the example, if the target fluorophore is the Alexa Fluor 488, using the Blue Laser and the GFP channel, 52.8% of the Alexa Fluor 488 emission spectra is captured. By reviewing the table horizontally and using the Blue Laser and the GFP channel or emission filter, the fluorescence emission from the GFP fluorophore is also captured with Alexa Fluor 488. In this example, the GFP signal is 76.1% of the Alexa Fluor 488 signal (the target for this channel).

Additionally, the amount of Alexa Fluor 488 spectral overlapping into the row's target fluorophore is displayed when reviewing the table vertically. In the example below the amount of the Alexa Fluor 488 emission signal using the RFP emission filter is captured from the Blue Laser. When using the Blue Laser and the RFP emission filter, the amount of Alexa Fluor 488 signal captured is 40.5% of the intended or Alexa Fluor 555 signal.

Changes to the fluorophores used can be made in the Spillover Table that result in changes in the Chart, resulting in an easy visual confirmation of the changes.