Pulsed-multiline excitation for color-blind fluorescence detection

A method of identifying sample components comprising:

• (a) preparing a sample comprising sample components, a first dye and a second dye;
• (b) placing the sample in the beam path of a first excitation line and a second excitation line;
• (c) sequentially firing the first excitation line and the second excitation line;
• (d) collecting fluorescence signals from the samples as a function of time;
• and
• (e) sorting the fluorescence by each excitation line's on-time window,
• wherein the sample components are identified.

A method of identifying sample components comprising:

• (a) obtaining a biological sample;
• (b) labeling said sample with one or more fluorophores;
• (c) separating components of said sample;
• and
• (d) detecting said sample components with a device wherein said device comprises:
• one or more lasers configured to emit two or more excitation lines, each excitation line having a different excitation wavelength;
• a timing circuit coupled to the one or more lasers and configured to fire the two or more excitation lines sequentially according to a timing program to produce time-correlated fluorescence emission signals from the sample;
• and
• a non-dispersive detector positioned to collect the time-correlated fluorescence emission signals;
• wherein said detector collects time correlated data from said sample comprising fluorescent emissions of the sample as a result of irradiation by the one or more excitation lines.

A device comprising:

• (a) one or more lasers having two or more excitation lines;
• (b) one or more beam steering mirrors wherein said excitation lines each strike said mirrors;
• (c) a first prism, wherein said two or more excitation lines strike one surface and exit from a second surface of said first prism;
• and
• (d) a second prism at an angle relative to said first prism, wherein said two or more excitation lines strike one surface of said second prism after exiting said first prism and exit said second prism,
• wherein said two or more excitation lines are substantially colinear or coaxial after exiting said second prism.

A method of illuminating a sample comprising:

• (a) steering two or more excitation lines onto a first surface of a first prism;
• (b) steering two or more excitation lines from the second surface of said first prism to a first surface of a second prism;
• wherein said second prism is angled about 45° from said first prism;
• (c) steering said two or more excitation lines onto a sample after exiting second surface of said second prism, wherein said two or more excitation lines are substantially colinear or coaxial after exiting said second prism.