Method for multiplexed capillary electrophoresis signal cross-talk correction
View Patent ↗The present invention provides a simple method to correct cross-talk, after the data have been generated. Adjacent signals are simply subtracted from the original observed signal with a repeating process. The data processing is stopped when a predefined condition is met. By this technique, cross-talk can be reduced from >5% to less than 0.1%. And as an additional advantage, this method provides a way to correct the cross-talk without the need to know which peaks are caused by the adjacent capillary signal.
1. A method of correcting multiplexed capillary electrophoresis (CE) electropherograms for cross-talk, comprising:
a) preparing an observed signal electropherogram(n) for a selected capillary of a multi-capillary electrophoresis system;
b) determining if a capillary exists z capillaries away from said selected capillary, and if so,
i) preparing an observed signal (CE) electropherogram(n+z) for a capillary located z capillaries away from said selected capillary (n)
ii) selecting a value that is greater than zero or less than or equal to a preset C % value that is less than or equal to 100 and designating this as A % value
iii) multiplying the A %-value onto each point of said electropherogram(n+z) to result in a modified signal electropherogram (n+z)-mod;
iv) subtracting said modified signal electropherogram(n+z)-mod from said signal electropherogram(n), resulting in a modified signal electropherogram(n)-mod;
v) selecting a new A %-value;
vi) iterating the process steps III)-through v) until the A % value is the largest possible value between zero and said C % value without giving any values in electropherogram(n)-mod less than a predefined B value
vii) preparing a corrected multiplexed capillary electrophoresis electropherogram by replacing each value of said signal electropherogram(n) with each corresponding value of electropherogram(n)-mod.
2. The method of claim 1 , wherein the said signal electropherogram(n) is corrected for cross-talk for each adjacent capillary located z=1 and z=−1 capillaries away from said selected capillary.
3. The method of claim 1 , wherein the said signal electropherogram(n) is corrected for cross-talk for each capillary located z capillaries away, from said selected capillary where z has the values −3, −2, −1, 1, 2, 3.
4. The method of claim 1 , wherein the said signal electropherogram(n) is corrected for cross-talk for any capillaries located z capillaries away, from said selected capillary where the integer values of z, excluding zero may be selected from the integer values −10 to +10.
5. The method of claim 1 , wherein the preparation of electropherograms includes a baseline correction step resulting in electropherograms with no negative values.
6. The method of claim 1 , wherein the A % value is increased or decreased in value upon each iteration step v.
7. The method of claim 1 , wherein the A % value in step v is calculated or predicted based on the magnitude of cross-talk correction from the previous iteration.
8. The method of claim 1 , wherein the A % value in step v is randomly selected between zero percent and a preset C-value.
9. The method of claim 1 wherein the A %-value is within the range of 0.05% to 99%.
10. The method of claim 1 wherein the said predefined B value is between 0.1% and −99% of the largest signal intensity of adjacent signal electropherogram(n+z).
11. The method of claim 1 , wherein the final maximum A % value that does not give any negative values in electropherogram(n)-mod less than a predefined B value is determined to within the nearest 1%.
12. The method of claim 1 , wherein the final maximum A % value that does not give any negative values in electropherogram(n)-mod less than a predefined B value in electropherogram(n)-mod is determined to within the nearest 0.1%.
13. The method of claim 1 , wherein the final maximum A % value that does not give any negative values in electropherogram(n)-mod less than a predefined B value is determined to within a range of the nearest 1% to the nearest 0.1%.
14. The method of claim 1 wherein the C value is between 1% and 99.9%.