Signal generation and mixing electronics for frequency-domain lifetime and spectral fluorometry

A system for fluorescence lifetime and spectral measurements, comprising:

• means for generating a driving/reference signal, said driving/reference signal means modulates the amplitude and/or the frequency of the driving/reference signal over time;
• means for generating a mixing signal, said mixing signal means modulates the amplitude and/or the frequency of the mixing signal over time,
• means for generating an excitation signal, the driving/reference signal drives said excitation signal means;
• means for detecting the emitted signal;
• means for mixing the emitted signal with the driving/reference signal to produce the processor reference signal;
• means for mixing the emitted signal with the mixing signal to produce the data signal;
• and
• a processor that extracts the fluorescence lifetime and fluorescence spectrum of the emitted signal from the comparison of the processor reference signal with the data signal using a chemometric analysis.

A method for measuring the fluorescence lifetime and the fluorescence spectrum, comprising the following steps:

• generating a driving/reference signal and modulating the amplitude and/or the frequency of the driving/reference signal over time;
• generating a mixing signal and modulating the amplitude and/or the frequency of the mixing signal over time;
• generating an excitation signal from the driving/reference signal;
• detecting the emitted signal,
• mixing the emitted signal with the driving/reference signal and producing the processor reference signal;
• mixing the emitted signal with the mixing signal producing the data signal;
• and
• extracting the fluorescence lifetime and fluorescence spectrum of the emitted signal from the comparison of the processor reference signal with the data signal to measure using a chemometric analysis.

A method of producing an apparatus for fluorescence lifetime and spectral measurements, comprising:

• providing a driving/reference signal generator that generates a driving/reference signal, said driving/reference signal is amplitude and/or frequency modulated over time;
• providing a mixing signal generator that generates a mixing signal, said mixing signal is amplitude and/or frequency modulated over time;
• coupling an excitation signal generator that generates an excitation signal and a reference signal to said driving/reference generator;
• providing a signal detector that detects the emitted signal;
• coupling a first mixer to said excitation signal generator, said mixer mixes the emitted signal with the driving/reference signal to produce the processor reference signal,
• coupling a second mixer to said mixing signal generator, said mixer mixes the emitted signal with the mixing signal to produce the data signal;
• and
• coupling a processor to said first mixer and said second mixer, said processor extracts the fluorescence lifetime and fluorescence spectrum of the emitted signal from the comparison of the processor reference signal with the data signal using a chemometric analysis.

A program storage device readable by a computer, tangibly embodying a program of instructions executable by the computer to perform method steps for a method for measuring the fluorescence lifetime and the fluorescence spectrum, comprising the following method steps:

• generating a driving/reference signal and modulating the amplitude and/or the frequency of the driving/reference signal over time;
• generating a mixing signal and modulating the amplitude and/or the frequency of the mixing signal over time;
• generating an excitation signal from the driving/reference signal;
• detecting the emitted signal;
• mixing the emitted signal with the driving/reference signal and producing the processor reference signal;
• mixing the emitted signal with the mixing signal producing the data signal;
• and
• extracting the fluorescence lifetime and fluorescence spectrum of the emitted signal from the comparison of the processor reference signal with the data signal to measure using a chemometric analysis.