Method and apparatus for simultaneous nonlinear excitation and detection of different chromophores across a wide spectral range using ultra-broadband light pulses and time-resolved detection

A method for simultaneous multicolor nonlinear imaging of multiple chromophores of different types in a sample, the method comprising:

• a) emitting ultrashort broadband light pulses from a few-cycle laser system for excitation of the multiple chromophores;
• b) focusing the ultrashort broadband light pulses in a sample plane of the sample;
• c) controlling the focused ultrashort broadband light pulses temporal duration in the sample plane;
• d) preventing the ultrashort broadband light pulses from directly reaching a detection system;
• e) measuring nonlinear signals generated in the sample plane;
• f) scanning the excitation and measurement on individual points (1D), or point-by-point in the sample plane (2D), or point-by-point across focal planes (3D);
• g) synchronizing the measurement of the non-linear signals generated in the sample plane to a detector in order to acquire time resolved nonlinear signal information on photon arrival time histograms from the sample plane;
• h) processing the time resolved nonlinear signal information;
• i) analyzing the time resolved non-linear signal information for determination of decay parameters associated to the photon arrival time histograms in the individual points (1D), or point-by-point in the sample plane (2D), or point-by-point across focal planes (3D).