Spectral deconvolution of fluorescent markers

An imaging system for determining a relative concentration of a specific fluorophore associated with an object, comprising:

• (a) a collection lens disposed so that light traveling from the object passes through the collection lens and travels along a collection path;
• (b) a dispersing component disposed in the collection path so as to receive the light that has passed through the collection lens, dispersing the light into a plurality of light beams, each light beam being directed away from the dispersing component in a different predetermined direction;
• (c) an imaging lens disposed to receive the light beams from the dispersing component, thereby producing an image from each different one of the light beams at a different predetermined location;
• (d) a detector disposed to receive the plurality of images produced by the imaging lens, the detector producing an output signal;
• and
• (e) means for processing the output signal to:
• (i) separate the output signal into portions corresponding to different fluorophores;
• and
• (ii) derive an amplitude for each portion of the output signal that corresponds to different fluorophores, the amplitude for each fluorophore corresponding to a relative concentration for that fluorophore.

A method for processing an electrical signal containing data from at least two sources, to separate the electrical signal into components corresponding to each of the at least two sources, comprising the steps of:

• (a) obtaining the electrical signal containing data from at least two sources;
• (b) processing the electrical signal to separate the electrical signal into portions corresponding to each of the at least two sources, said processing including at least one of:
• (i) utilizing a nonlinear conjugation, said nonlinear conjugation reducing the number of free parameters to be optimized by using data collected from an optimization calibration individually executed for each source;
• and
• (ii) solving a set of linear equations corresponding to a data set defined by the sources;
• and
• (c) deriving an amplitude for each portion of the electrical signal that corresponds to one of the at least two sources.

A method for processing an electrical signal containing data from at least two sources, to separate the electrical signal into components corresponding to each of the at least two sources, comprising the steps of:

• (a) obtaining the electrical signal containing data from at least two sources;
• (b) converting the electrical signal into a two-dimensional digital representation;
• (c) identifying portions of the two-dimensional digital representation that correspond to each of the at least two sources;
• and
• (d) performing a digital optimization to derive an amplitude for each portion of the two-dimensional digital representation that corresponds to one of the at least two sources, by fitting the two-dimensional digital representation to a parameterized model based on known data corresponding to each of the at least two sources.