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. Author manuscript; available in PMC: 2020 Sep 3.
Published in final edited form as: Methods Mol Biol. 2012;833:153–176. doi: 10.1007/978-1-61779-477-3_11

Fig. 1.

Fig. 1.

Basics of FRAP, (a) Shown is a nucleus (solid black outline) containing fluorescently tagged proteins (solid gray circles). Small arrows indicate protein motion. FRAP is performed by photobleaching a region (dashed circle) of the nucleus. During the recovery phase, bleached molecules (black circles) move out of the bleach zone and unbleached molecules move in. The rate of fluorescence recovery can be measured by estimating the average fluorescent intensity inside the measurement area (dashed circle) as a function of time, generating a FRAP curve. (b) The FRAP curve shows observational photobleaching, i.e., the fluorescence intensity decreases over time due to unintentional photobleaching during all periods of observation. The bleach depth is defined here as the initial measured value of the FRAP curve immediately after the photobleach.