Abstract: A system and method for fluorescence excitation and detection having distinct optical paths is disclosed. A system for detecting fluorescence comprises a light source that emits an excitation light into an illumination tube; a plurality of collection optics located around an aperture in the illumination tube for collecting fluorescence; and a detector for determining the amount of fluorescence. A method for detecting fluorescence comprises emitting an excitation light from a light source into an illumination tube; directing the excitation light to an excitation filter; illuminating a sample with the excitation light to generate an emission light; and detecting the optical characteristics of the emission light using a plurality of collection optics located around the illumination tube.

Abstract: The present invention provides systems and methods for normalizing data in nucleic acid amplification procedures. In general, the methods include: obtaining input from a user regarding a number of normalizer measurements to use in calculations; obtaining raw fluorescence measurements of one or more probe(s) over multiple cycles of the procedure; obtaining normalizer measurement(s); and normalizing the probe data using the normalizer measurement(s) specified by the user. The present invention also discloses software and a computer readable medium for carrying method steps of the invention. Reduced cycle times, decreased variability, and/or lower Ct values are obtained as a result of the normalization.