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(a) The algorithm starts by locating a bead close to the center of the field of view. (b) Next, the XY stage translates the bead in a random direction (+X, −X, +Y, −Y) for a total of 100 steps at 60 stage ticks each. (c) These translations provide four separate calibration factors, one for each direction of motion. (d) The final calibration routine uses large translations across the entire field of view and is repeated until the accuracy limit is reached.
