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. 1997 Jun 30;137(7):1459–1468. doi: 10.1083/jcb.137.7.1459

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Typical FRAP curves and associated theoretical recovery curves obtained from a rhodamine-labeled antibody undergoing Brownian diffusion in dilute solution (∼0.5 mg/ml antibody) for smaller (diamonds) and larger (squares) spots. The bleach pulse was 200 μs in duration, and data were collected in 100-μs increments. The upper curve represents an average of 1,000 experiments and the lower an average of 3,000 experiments, and each was smoothed using an 11-point fit to a fourth-order polynomial. The fluorescence recovers almost completely, indicating that all molecules are mobile. In addition, the recovery time increases with increasing spot size.