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A Simple and Sensitive Method to Quantify Biodegradable Nanoparticle Biodistribution using Europium Chelates.

Crawford L, Higgins J, Putnam D - Sci Rep (2015)

Bottom Line: The TRF of the nanoparticles was found to diminish as a second order function in the presence of serum and tissue compositions interfered with the europium signal.Both phenomena were corrected by linearization of the signal function and calculation of tissue-specific interference, respectively.Overall, the method is simple and robust with a detection limit five times greater than standard fluorescent probes.

View Article: PubMed Central - PubMed

Affiliation: School of Chemical and Biomolecular Engineering, Cornell University, Ithaca NY.

ABSTRACT
The biodistribution of biodegradable nanoparticles can be difficult to quantify. We report a method using time resolved fluorescence (TRF) from a lanthanide chelate to minimize background autofluorescence and maximize the signal to noise ratio to detect biodegradable nanoparticle distribution in mice. Specifically, antenna chelates containing europium were entrapped within nanoparticles composed of polylactic acid-polyethylene glycol diblock copolymers. Tissue accumulation of nanoparticles following intravenous injection was quantified in mice. The TRF of the nanoparticles was found to diminish as a second order function in the presence of serum and tissue compositions interfered with the europium signal. Both phenomena were corrected by linearization of the signal function and calculation of tissue-specific interference, respectively. Overall, the method is simple and robust with a detection limit five times greater than standard fluorescent probes.

No MeSH data available.


Characteristics of PLA-PEG nanoparticles doped with europium chelate.(a) representative structure of Eu(NTA)3 chelate. (b) excitation and emission spectra for 1 mg/mL nanoparticle suspension in PBS (solid lines) and 0.1 mg/mL chelate solution in acetone (dotted lines) under time resolved conditions.
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f1: Characteristics of PLA-PEG nanoparticles doped with europium chelate.(a) representative structure of Eu(NTA)3 chelate. (b) excitation and emission spectra for 1 mg/mL nanoparticle suspension in PBS (solid lines) and 0.1 mg/mL chelate solution in acetone (dotted lines) under time resolved conditions.

Mentions: Polylactic acid–mono methoxy polyethylene glycol (PLA-PEG) diblock copolymers were synthesized by ring opening polymerization. Mn was consistent between both GPC and 1H NMR (21,000 and 20,600, respectively). Nanoparticle fabrication yielded particles 106 ± 6.5 nm in diameter and −1.45 ± 0.25 mV surface charge with a polydispersity of 0.079 ± 0.03. Excitation and emission spectra for the particles and chelate (Fig. 1), show maximum excitation at 340 nm and a maximum emission at 610 nm. Both the unencapsulated chelate and the chelate-containing nanoparticles show a large Stokes shift, which is a hallmark characteristic of europium chelates. Release studies of europium chelate from PLA-PEG nanoparticles (Figure S1) show no detectable chelate release over nine hours.


A Simple and Sensitive Method to Quantify Biodegradable Nanoparticle Biodistribution using Europium Chelates.

Crawford L, Higgins J, Putnam D - Sci Rep (2015)

Characteristics of PLA-PEG nanoparticles doped with europium chelate.(a) representative structure of Eu(NTA)3 chelate. (b) excitation and emission spectra for 1 mg/mL nanoparticle suspension in PBS (solid lines) and 0.1 mg/mL chelate solution in acetone (dotted lines) under time resolved conditions.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561907&req=5

f1: Characteristics of PLA-PEG nanoparticles doped with europium chelate.(a) representative structure of Eu(NTA)3 chelate. (b) excitation and emission spectra for 1 mg/mL nanoparticle suspension in PBS (solid lines) and 0.1 mg/mL chelate solution in acetone (dotted lines) under time resolved conditions.
Mentions: Polylactic acid–mono methoxy polyethylene glycol (PLA-PEG) diblock copolymers were synthesized by ring opening polymerization. Mn was consistent between both GPC and 1H NMR (21,000 and 20,600, respectively). Nanoparticle fabrication yielded particles 106 ± 6.5 nm in diameter and −1.45 ± 0.25 mV surface charge with a polydispersity of 0.079 ± 0.03. Excitation and emission spectra for the particles and chelate (Fig. 1), show maximum excitation at 340 nm and a maximum emission at 610 nm. Both the unencapsulated chelate and the chelate-containing nanoparticles show a large Stokes shift, which is a hallmark characteristic of europium chelates. Release studies of europium chelate from PLA-PEG nanoparticles (Figure S1) show no detectable chelate release over nine hours.

Bottom Line: The TRF of the nanoparticles was found to diminish as a second order function in the presence of serum and tissue compositions interfered with the europium signal.Both phenomena were corrected by linearization of the signal function and calculation of tissue-specific interference, respectively.Overall, the method is simple and robust with a detection limit five times greater than standard fluorescent probes.

View Article: PubMed Central - PubMed

Affiliation: School of Chemical and Biomolecular Engineering, Cornell University, Ithaca NY.

ABSTRACT
The biodistribution of biodegradable nanoparticles can be difficult to quantify. We report a method using time resolved fluorescence (TRF) from a lanthanide chelate to minimize background autofluorescence and maximize the signal to noise ratio to detect biodegradable nanoparticle distribution in mice. Specifically, antenna chelates containing europium were entrapped within nanoparticles composed of polylactic acid-polyethylene glycol diblock copolymers. Tissue accumulation of nanoparticles following intravenous injection was quantified in mice. The TRF of the nanoparticles was found to diminish as a second order function in the presence of serum and tissue compositions interfered with the europium signal. Both phenomena were corrected by linearization of the signal function and calculation of tissue-specific interference, respectively. Overall, the method is simple and robust with a detection limit five times greater than standard fluorescent probes.

No MeSH data available.