Methods of identifying biological targets and instrumentation to identify biological targets

A method of detecting a biological target, comprising:

• providing a first bead and a second bead, wherein different wavelengths of emission energy are emitted from the first bead and the second bead, wherein the emission from the first bead is at a first wavelength and the emission from the second bead is at a second wavelength, wherein the first wavelength and the second wavelength are detectably distinguishable, wherein the first bead includes a first biomolecule having an affinity for a first binding site of a biological target, wherein the second bead includes a second biomolecule having an affinity for a second binding site of the biological target, and wherein the biological target includes the first binding site and the second binding site;
• mixing the first bead and the second bead with a sample, wherein if the sample includes the biological target, the first bead binds to the first binding site of the target molecule and the second bead binds to the second binding site of the target molecule to form a first bead/biological target/second bead complex;
• exposing the first bead/biological target/second bead complex to a single irradiation source emitting radiation at a single wavelength;
• and
• simultaneously detecting the emission energy emitted from each of the first bead and the second bead, and wherein simultaneously detecting the emission from the first bead and the second bead is correlated to the detection of the first bead /biological target/second bead complex.

A single-molecule detection system comprising:

• a channel for flowing a fluid that includes a fluorescent/biological target complex, wherein the fluorescent/biological target complex emits at least two radiation energy signals upon irradiation by a single wavelength of radiation energy, wherein each radiation energy signal is at a distinguishably different wavelength;
• an irradiation and optic system including an irradiation source configured to direct a single wavelength of an irradiation beam onto a portion of the channel, wherein the irradiation and optic system is configured to direct the at least two emitted radiation energy signals from the fluorescent/biological target complex to the detector and co-incidence system;
• a detector and co-incidence system including at least two detectors, wherein each detector includes a filter that allows only a wavelength for one of the radiation energy signals emitted from the fluorescent/biological target complex to pass through each of the filters, wherein the detector and co-incidence system includes a co-incidence detector interfaced to one of the detectors and a delay generator interfaced to the another detector, wherein the delay generator and the co-incidence detector are directly interfaced, wherein the co-incidence detector is configured to determine if co-incident signals are emitted from the fluorescent/biological target complex;
• and
• an analysis system in communication with the co-incidence detector, wherein the analysis system receives data from the co-incidence detector if the detectors simultaneously receive radiation energy signals emitted from the fluorescent/biological target complex.