Two-photon microscopy and spectroscopy of lanthanide bioprobes.
The linear and non-linear photophysical properties of tris-dipicolinate europium and terbium complexes (absorption, emission, lifetime, luminescence induced by two-photon absorption) are studied in the crystalline state as well as in protein derivative crystals and compared to those in solution. Upon laser irradiation at 532 nm, luminescence of terbium is induced by a two-photon antenna effect, whereas luminescence of europium results from one-photon absorption in forbidden f-f transitions. Finally, linear and two-photon microscopy imaging experiments on biological and bio-inspired crystals are performed. These first proof-of-concept experiments open the way for the development of time-resolved non-linear microscopy that should combine the advantages of lanthanide luminescence (long lifetime, sharp emission bands, insensitivity to oxygen) with those of confocal biphotonic excitation (near-IR excitation, 3D resolution and reduced photodamage).
D'Aléo
Anthony
A
Laboratoire de Chimie, ENS-Lyon, CNRS, 46 Allée d'Italie, 69364 Lyon, France.
Pompidor
Guillaume
G
Elena
Bénédicte
B
Vicat
Jean
J
Baldeck
Patrice L
PL
Toupet
Loïc
L
Kahn
Richard
R
Andraud
Chantal
C
Maury
Olivier
O
D001703:Biophysics / Q000379:methods; D015374:Biosensing Techniques; D002627:Chemistry, Physical / Q000379:methods; D003460:Crystallization; D005063:Europium / Q000737:chemistry; D028581:Lanthanoid Series Elements / Q000494:pharmacology*; D008027:Light; D009682:Magnetic Resonance Spectroscopy; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D018613:Microscopy, Confocal; D010777:Photochemistry; D017785:Photons; D013055:Spectrophotometry, Infrared / Q000379:methods; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013725:Terbium / Q000737:chemistry; D013997:Time Factors
D028581:Lanthanoid Series Elements; D013725:Terbium; D005063:Europium