Journal
BIOCONJUGATE CHEMISTRY
Volume 22, Issue 7, Pages 1438-1450Publisher
AMER CHEMICAL SOC
DOI: 10.1021/bc2002506
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Funding
- National Institutes of Health [1 R15 EB008858-01, 1 R01 CA125255]
- U.S. Civilian Research and Development Foundation [UKB2-2923-KV-07]
- Ministry of Education and Science of Ukraine [M/49-2008]
- National Science Foundation [CHE-0840431, CHE-0832622]
- Florida Department of Health [10BD-11]
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A two-photon absorbing (2PA) and aggregation-enhanced near-infrared (MR) emitting pyran derivative, encapsulated in and stabilized by silica nanopartides (SiNPs), is reported as a nanoprobe for two-photon fluorescence microscopy (2PFM) bioimaging that overcomes the fluorescence quenching associated with high chromophore loading. The new SiNP probe exhibited aggregate-enhanced emission producing nearly twice as strong a signal as the unaggregated dye, a 3-fold increase in two-photon absorption relative to the DFP in solution, and approximately 4-fold increase in photostability. The surface of the nanoparticles was functionalized with a folic acid (FA) derivative for folate-mediated delivery of the nanoprobe for 2PFM bioimaging. Surface modification of SiNPs with the FA derivative was supported by zeta potential variation and H-1 NMR spectral characterization of the SiNPs as a function of surface modification. In vitro studies using HeLa cells expressing a folate receptor (FR) indicated specific cellular uptake of the functionalized nanopartides. The nanoprobe was demonstrated for FR-targeted one-photon in vivo imaging of HeLa tumor xenograft in mice upon intravenous injection of the probe. The FR-targeting nanoprobe not only exhibited highly selective tumor targeting but also readily extravasated from tumor vessels, penetrated into the tumor parenchyma, and was internalized by the tumor cells. Two-photon fluorescence microscopy bioimaging provided three-dimensional (3D) cellular-level resolution imaging up to 350 mu m deep in the HeLa tumor.
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