Fluorescence resonance energy transfers measurements on cell surfaces via fluorescence polarization.
A method has been developed for the determination of the efficiency of fluorescence resonance energy transfer efficiency between moieties located on cell surfaces by performing individual cell fluorescence polarization (FP) measurements. The absolute value of energy transfer efficiency (E) is calculated on an individual cell basis. The examination of this methodology was carried out using model experiments on human T lymphocyte cells. The cells were labeled with fluorescein-conjugated Concanavalin A (ConA) as donor, or rhodamine-conjugated ConA as acceptor. The experiments and results clearly indicate that determination of E via FP measurements is possible, efficient, and more convenient than other methods.
The Biophysical Interdisciplinary Jerome Schottenstein Center for the Research and Technology of the Cellome Physics Department, Bar-Ilan University, Ramat-Gan 52900, Israel.
Meir
M
Cohen-Kashi
Sergey
S
Moshkov
Naomi
N
Zurgil
Mordechai
M
Deutsch
D055592:Biophysical Phenomena; D001703:Biophysics; D002462:Cell Membrane / Q000502:physiology*; D002469:Cell Separation; D003208:Concanavalin A / Q000494:pharmacology; D005434:Flow Cytometry; D005454:Fluorescence Polarization / Q000295:instrumentation / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008962:Models, Theoretical; D011485:Protein Binding; D012235:Rhodamines / Q000494:pharmacology; D013601:T-Lymphocytes / Q000166:cytology
D012235:Rhodamines; D003208:Concanavalin A