Translator Disclaimer
13 February 2012 "Sizing" the oligomers of Azami Green fluorescent protein with FCS and antibunching
Author Affiliations +
Abstract
Fluorescent proteins are invaluable molecules in fluorescence microscopy and spectroscopy. The size and brightness of fluorescent proteins often dictates the application they may be used for. While a monomeric protein may be the least perturbative structure for labeling a protein in a cell, often oligomers (dimers and tetramers) of fluorescent proteins can be more stable. However, from a quantitative microscopy standpoint, it is important to realize the photophysical properties of monomers do not necessarily multiply by their number when they form oligomers. In this work we studied oligomerization states of the Azami Green (AG) protein with fluorescence correlation spectroscopy (FCS) and photon antibunching or photon pair correlation spectroscopy (PPCS). FCS was used to measure the hydrodynamic size of the oligomers, whereas antibunching was used to count the number of fluorescent emitters in the oligomers. The results exhibited that the dimers of AG were single emitters and the tetramers were dual-emitters, indicative of dipole-dipole interactions and energy transfer between the monomeric units. We also used these methods to estimate the number of fluorescent proteins displayed on T7 phage molecules.
© (2012) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Jamshid Temirov, James H. Werner, Peter M. Goodwin, and Andrew R. M. Bradbury ""Sizing" the oligomers of Azami Green fluorescent protein with FCS and antibunching", Proc. SPIE 8228, Single Molecule Spectroscopy and Superresolution Imaging V, 82280J (13 February 2012); https://doi.org/10.1117/12.906843
PROCEEDINGS
9 PAGES


SHARE
Advertisement
Advertisement
Back to Top