McNally James G
Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, Bethesda, Maryland 20892, USA.
Methods Cell Biol. 2008;85:329-51. doi: 10.1016/S0091-679X(08)85014-5.
Fluorescence recovery after photobleaching (FRAP) reveals the dynamics of fluorescently tagged molecules within live cells. These molecular dynamics are governed by diffusion of the molecule and its in vivo binding interactions. As a result, quantitative estimates of the association and dissociation rates of binding can be extracted from the FRAP. This chapter describes a systematic procedure to acquire the FRAP data, and then fit it with appropriate mathematical models to estimate in vivo association and dissociation rates of binding. Also discussed are the applicability and limitations of the models, the utility of the estimated parameters, and the prospects for increased accuracy and confidence in the estimates.
光漂白后荧光恢复（FRAP）揭示了活细胞内荧光标记分子的动力学。这些分子动力学受分子扩散及其体内结合相互作用的支配。因此，可以从FRAP中提取结合的缔合和解离速率的定量估计值。本章描述了获取FRAP数据的系统程序，然后将其与适当的数学模型拟合，以估计体内结合的缔合和解离速率。还讨论了模型的适用性和局限性、估计参数的效用，以及提高估计准确性和可信度的前景。
