Electrophoretic analysis system having in-situ calibration
View Patent ↗An electrophoretic system having a plurality of separation lanes is provided with an automatic calibration feature in which each lane is separately calibrated. For each lane, the calibration coefficients map a spectrum of received channel intensities onto values reflective of the relative likelihood of each of a plurality of dyes being present. Individual peaks, reflective of the influence of a single dye, are isolated from among the various sets of detected light intensity spectra, and these can be used to both detect the number of dye components present, and also to establish exemplary vectors for the calibration coefficients which may then be clustered and further processed to arrive at a calibration matrix for the system. The system of the present invention thus permits one to use different dye sets to tag DNA nucleotides in samples which migrate in separate lanes, and also allows for in-situ calibration with new, previously unused dye sets.
1. A method for calibrating a separation system having at least one separation lane, comprising:
clustering a plurality of sets of detector signals into a number n categories by using at least one clustering criterion, wherein:
each set of detector signals corresponds to light intensities at each of a plurality of different wavelengths detected from each sample of a plurality of samples separated along the at least one separation lane; and
the step of clustering comprises normalizing at least some detector signals of each set of detector signals by a respective normalization value to prepare a plurality of sets of normalized detector signals, each set comprising normalized detector signals;
comparing corresponding normalized detector signals of different sets of normalized detector signals; and
clustering normalized sets of detector signals that do not have compared corresponding normalized detector signals differing by more than the clustering criterion; and
calibrating the separation system based on the clusters.
2. A separation system having at least one separation lane, comprising:
a processor configured to cluster a plurality of sets of detector signals into a number n categories by using at least one clustering criterion, wherein:
each set of detector signals corresponds to light intensities at each of a plurality of different wavelengths detected from each sample of a plurality of samples separated along the at least one separation lane; and
the processor is configured to at least:
normalize at least some detector signals of each set of detector signals by a respective normalization value to prepare a plurality of sets of normalized detector signals, each set comprising normalized detector signals;
compare corresponding normalized detector signals of different sets of normalized detector signals; and
cluster normalized sets of detector signals that do not have compared corresponding normalized detector signals differing by more than the clustering criterion; and
calibrate the separation system based on the clusters.