High-fidelity hydrophilic probe for two-photon fluorescence lysosomal imaging.
The synthesis and characterization of a novel two-photon-absorbing fluorene derivative, LT1, selective for the lysosomes of HCT 116 cancer cells, is reported. Linear and nonlinear photophysical and photochemical properties of the probe were investigated to evaluate the potential of the probe for two-photon fluorescence microscopy (2PFM) lysosomal imaging. The cytotoxicity of the probe was investigated to evaluate the potential of using this probe for live two-photon fluorescence biological imaging applications. Colocalization studies of the probe with commercial Lysotracker Red in HCT 116 cells demonstrated the specific localization of the probe in the lysosomes with an extremely high colocalization coefficient (0.96). A figure of merit was introduced to allow comparison between probes. LT1 has a number of properties that far exceed those of commercial lysotracker probes, including higher two-photon absorption cross sections, good fluorescence quantum yield, and, importantly, high photostability, all resulting in a superior figure of merit. 2PFM was used to demonstrate lysosomal tracking with LT1.
Wang
Xuhua
X
Department of ChemistryUniversity of Central Florida, Orlando, Florida 32816, USA.
Nguyen
Dao M
DM
Yanez
Ciceron O
CO
Rodriguez
Luis
L
Ahn
Hyo-Yang
HY
Bondar
Mykhailo V
MV
Belfield
Kevin D
KD
D000814:Aniline Compounds / Q000138:chemical synthesis* / Q000737:chemistry; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D005449:Fluorenes / Q000138:chemical synthesis* / Q000737:chemistry; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D008247:Lysosomes / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D017785:Photons; D011789:Quantum Theory
C554358:4,4'-(9,9-di(2,5,8,11,14,17,20,23,26,29-decaoxahentriacontan-31-yl)-9H-fluorene-2,7-diyl)bis(ethyne-2,1-diyl)bis(N,N-dimethylaniline); D000814:Aniline Compounds; D000970:Antineoplastic Agents; D005449:Fluorenes; D005456:Fluorescent Dyes