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Each color now is mixed and created for each individual spot.
Normalization of each individual spot was performed according to the total quantity of the valid spots in each gel, after subtraction of the background values.
In practice, when we loaded 6×106 cells on a 1-chamber slide, the average number of cells attaching to each individual spot was 897±106 (loose attached) (Figure 1C, left and middle panel; Figure 1D).
Local background correction was applied to each individual spot.
A correction factor can then be calculated from a generalized linear model for each individual spot.
This normalization included correction for the random error, with the median Cy3 signal for each individual spot.
Similar(42)
Where multiple protein products were identified from a single gene the correlation was calculated using the transcriptomic data for that gene versus proteomic data for each individual protein spot (i.e. multiple spots were not averaged or summed), because of the possibility that not all spots for any one gene product were detected.
Individual spot volumes for each gel were normalized relative to the total spot volume of that gel.
Each individual hot spot was classified as metastasis or not by an artificial neural network (ANN).
Each individual analyte spot was positioned directly under the EF-LESA probe and sampled as follows: (i) voltages V1, V2, and V3 were initially set to zero.
Manual rotation of the sample plate on the cone allowed three separate laser shots to ablate an area of each individual matrix/analyte spot.
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