Visualizing the distribution of proteins and estimating their kinetic parameters using Virtual Photoactivated Fluorescence (VPAF)

Abstract

Photoactivated Fluorescence (PAF) and
Fluorescence Recovery After Photobleaching (FRAP) are two
inverse related fluorescence microscopy techniques used to
study the kinetic of nuclear proteins. In this paper, we will
propose and use a method, Virtual Photoactivated
Fluorescence (VPAF), based on the inverse relationship
between PAF and FRAP, to visualize and quantify the
dynamics of proteins within the cell nucleus. In particular, we
will visualize the heterogeneous distribution of splicing factors
throughout the cell nucleus after virtual photoactivation and
estimate kinetic parameters of linker histones using VPAF
data.