Clipping-assisted dual-fluorophore sensing

A method is provided for characterizing a biological sample having a plurality of fluorophores, including a red fluorophore and a blue fluorophore, comprises exciting the red fluorophore via absorption of a photon order of n by a single wavelength band of light that has longer wavelengths than a typical wavelength band of light known to excite the red fluorophore would have. The method further comprises exciting the blue fluorophore substantially via absorption of a photon order of n+1 by the single wavelength band of light. The method also comprises simultaneously detecting light emitted by the red fluorophore and the blue fluorophore. The method further comprises creating an image or a temporal series for sensing from the light detected in the plurality of orthogonal colors.