Systems and methods for detecting and analyzing polymers

A detection system for analyzing a polymer having three or more distinct fluorophores, each fluorophore having an excitation wavelength and a corresponding emission bandwidth, the detection system comprising:

• at least three different emitters that together create an excitation signal configured to excite each of the at least three fluorophores such that each fluorophore emits a distinct emission signal within its respective emission bandwidth that does not overlap any other of the distinct emission signals at normalized intensities above 70%;
• a polymer excitation zone that accepts the polymer, the polymer excitation zone being illuminated by the excitation signal of the at least three different emitters;
• a detector constructed and arranged to detect the distinct emission signal from each of the at least three fluorophores;
• and
• a polychroic mirror adapted to prevent excitation signals emitted from the at least three different emitters from reaching the detector.

A detection system for analyzing a polymer having three or more distinct fluorophores, each fluorophore having an excitation wavelength and a corresponding emission bandwidth, the detection system comprising:

• at least three different emitters that together create an excitation signal having wavelengths configured to excite each of a corresponding one of the at least three fluorophores such that each fluorophore emits a distinct emission signal within its respective emission bandwidth, each of the at least three emitters and corresponding at least three fluorophores selected from a group consisting of:
• an emitter emitting an excitation wavelength of 488 nm and a corresponding fluorophore having an emission maximum substantially located at 512 nm, an emitter emitting an excitation a wavelength of 532 nm and a corresponding fluorophore having an emission maximum substantially located at 575 nm, an emitter emitting an excitation wavelength of 633 nm and a corresponding fluorophore having an emission maximum substantially located at 665 nm, and an emitter emitting an excitation wavelength of 750 nm and a corresponding fluorophore having an emission maximum substantially located at 775 nm or 806 nm;
• a polymer excitation zone that accepts the polymer, the polymer excitation zone being illuminated by the excitation wavelengths of the at least three different emitters;
• a detector constructed and arranged to detect the distinct emission signal from each of the at least three fluorophores;
• and
• a polychroic mirror adapted to prevent excitation signals emitted from the at least three different emitters from reaching the detector.

A method of detecting emissions of three or more distinct fluorophores bound to a polymer, the method comprising:

• selecting three or more fluorophores, each of the fluorophores characterized by an excitation wavelength and a corresponding emission bandwidth, the emission bandwidth of each of the fluorophores not overlapping the emission bandwidth of any other of the fluorophores at normalized intensities above 70%;
• attaching the three or more fluorophores to the polymer in a sequence specific manner;
• illuminating each of the fluorophores with an excitation signal of multiple emitters, the excitation signal comprising excitation wavelengths of the corresponding fluorophores that cause each of the fluorophores to emit an emission signal within the emission bandwidth of the fluorophore;
• removing the excitation signal from each of the emission signals with a polychroic filter;
• and
• detecting and analyzing the emission signal of each of the fluorophores.

A method for analyzing a polymer, the method comprising:

• selecting at least three distinct fluorophores and an excitation signal emitted from at least three emitters, each of the at least three fluorophores having an excitation wavelength and a corresponding emission bandwidth, the excitation signal having excitation wavelengths that correspond to each of the at least three fluorophores such that each fluorophore emits a distinct emission signal having an emission maximum within its respective emission bandwidth, each of the at least three excitation wavelengths and corresponding at least three fluorophores selected from a group consisting of:
• an excitation wavelength of 488 nm and a corresponding fluorophore having an emission maximum substantially located at 512 nm, an excitation wavelength of 532 nm and a corresponding fluorophore having an emission maximum substantially located at 575 nm, an excitation wavelength of 633 nm and a corresponding fluorophore having an emission maximum substantially located at 665 nm, and an excitation wavelength of 750 nm and a corresponding fluorophore having an emission maximum substantially located at 775 nm or 806 nm;
• binding the at least three fluorophores to a polymer in a sequence specific manner;
• illuminating each of the fluorophores with the corresponding emitter at a common time;
• removing the excitation signal from each of the emission signals with a polychroic filter;
• and
• detecting and analyzing the emission signal of each of the fluorophores for analysis.