Fluorescence resonance energy transfer imaging via fluorescence polarization measurement.
Fluorescence resonance energy transfer (FRET) has become a widely used spectroscopic tool for detecting molecular interactions and molecular proximity in solution, as well as in membranes. On the other hand, fluorescence polarization (FP) is a convenient measure: ratiometric and simple to execute. This work presents a novel methodology for determining energy transfer efficiency (E) via FP measurement. The methodology is based on the fact that a donor's fluorescence lifetime is shortened due to FRET and, consequently, its FP increases. As a model, the present work evaluates the E between fluorescein and rhodamine conjugated ConA attached to the receptors in the lymphocyte membrane. It shows not only that FRET imaging via FP is possible, but also that it is inexpensive, simple to perform, conveniently adaptable to the commonly used fluorescent microscopy, and readily interpretable.
Cohen-Kashi
Meir
M
Bar Ilan University, Department of Physics, Biophysical Interdisciplinary Schottenstein Center for the Research and the Technology of the Cellome, Ramat Gan, Israel.
Namer
Yaniv
Y
Deutsch
Mordechai
M
D002460:Cell Line; D003208:Concanavalin A / Q000493:pharmacokinetics*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008859:Microscopy, Polarization / Q000379:methods*; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism*
D003208:Concanavalin A