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Multi-Photon Laser Scanning Microscopy (MPLSM) requires efficient two-photon absorbing fluorescent (TPF) probes. In particular, probes exhibiting bio-functionality are very attractive for MPLSM studies of biological samples. We have synthesized and studied a new class of TPF probes capable of caging metal ions, such as Ca+2 and Na+, which play an important role in neuronal mechanisms. The TPF probes are based on a tetraketo derivative with a symmetric Donor-Acceptor-Donor (D-A-D) structure. The donor is an azacrown moiety, which also serves as a metal ion-caging unit. We studied the linear and the non-linear spectroscopic properties of these TPF probes as a function of conjugation length and the size of the crown ring. We find that this new class of TPF probes possesses very large two-photon excitation cross-section coefficients (~1000GM) at near IR wavelengths as well as affinity to metal ions. In the presence of changing sodium ion concentration the dye spectra reveals four distinguishable forms and the TPF efficiency changes strongly. We therefore conclude that the dye can perform as a sensitive metal ion TPF probe.
Raz Gvishi,Garry Berkovic,Zvi Kotler,Pnina Krief,Lev Shapiro,Jacob T. Klug,Jacqueline Skorka, andVladimir Khodorkovsky
"Studies of new two-photon fluorescent probes suitable for multiphoton microscopy in biological settings", Proc. SPIE 5211, Nonlinear Optical Transmission and Multiphoton Processes in Organics, (3 November 2003); https://doi.org/10.1117/12.506766
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Raz Gvishi, Garry Berkovic, Zvi Kotler, Pnina Krief, Lev Shapiro, Jacob T. Klug, Jacqueline Skorka, Vladimir Khodorkovsky, "Studies of new two-photon fluorescent probes suitable for multiphoton microscopy in biological settings," Proc. SPIE 5211, Nonlinear Optical Transmission and Multiphoton Processes in Organics, (3 November 2003); https://doi.org/10.1117/12.506766