研究目的
To synthesize and characterize a series of single and multidye copolymerized silica nanoparticles with large Stokes shifts for versatile biosensing applications, including in vitro and in vivo imaging.
研究成果
The synthesized nanoparticles exhibit large Stokes shifts, high reproducibility, minimal dye leakage, enhanced fluorescence intensities, and good biocompatibility after surface modification. They show promise for multicolor biosensing and in vivo imaging applications, with potential for expansion to additional dye combinations and further functionalization.
研究不足
The study is limited to in vitro and proof-of-concept in vivo assessments; further in vivo testing is needed. The protein adsorption for biotin-PEG nanoparticles was slightly higher than for PEG-only nanoparticles, warranting further investigation. The method may require optimization for different dye species.
1:Experimental Design and Method Selection:
Nanoparticles were synthesized via the St?ber method with covalent copolymerization of dyes (FITC, Texas Red, and novel aminocyanine dye GC-1-23). Surface modification with PEG and biotin was performed for bioconjugation.
2:3). Surface modification with PEG and biotin was performed for bioconjugation. Sample Selection and Data Sources:
2. Sample Selection and Data Sources: Dye-silane conjugates were prepared and used in nanoparticle synthesis. Human red blood cells and serum proteins (HSA, IgG) were used for biocompatibility tests.
3:List of Experimental Equipment and Materials:
Includes Biotek Synergy H1 and H4 Microplate Readers, ZEISS Leo 906E Transmission Electron Microscope, Olympus BX51WI fluorescence microscope, Eppendorf pipettes, and various chemicals from Sigma Aldrich, Thermo Fisher Scientific, Bioclone Inc., Laysan Bio, and Chemodex.
4:Experimental Procedures and Operational Workflow:
Dye-silane conjugates were synthesized, nanoparticles were prepared via controlled TEOS addition, washed, and characterized for size, yield, quantum yield, and fluorescence. Surface modification involved PEGylation and biotinylation. Biocompatibility was assessed through hemolysis and protein adsorption studies.
5:Data Analysis Methods:
UV-visible and fluorescence spectroscopy, TEM imaging, fluorescence microscopy, and statistical analysis using software like Biotek Gen5, Microsoft Excel, FluorTools a|e software, and ImageJ.
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Leo 906E Transmission Electron Microscope
Leo 906E
ZEISS
Transmission electron microscopy for nanoparticle size characterization
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BX51WI Fluorescence Microscope
BX51WI
Olympus
Fluorescence microscopy imaging
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Amicon Ultra Centrifugal Filter Units
10 kDa
Thermo Fisher Scientific
Filtration and concentration of nanoparticle solutions
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Synergy H1 Microplate Reader
Synergy H1
Biotek
UV-visible and fluorescence spectroscopic measurements
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Synergy H4 Microplate Reader
Synergy H4
Biotek
UV-visible and fluorescence spectroscopic measurements
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X-Cite Series 120 Q Light Source
X-Cite Series 120 Q
X-Cite
Excitation light source for fluorescence microscopy
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Infinity 3S-iUR CCD Camera
Infinity 3S-iUR
Infinity
Image acquisition in fluorescence microscopy
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Eppendorf Pipette
10–100 μL or 100–1000 μL adjustable volume
Eppendorf
Precise volume delivery for experiments
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BcMag Streptavidin Magnetic Beads
1 μm
Bioclone Inc.
Binding and conjugation with biotinylated nanoparticles for proof-of-concept assays
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