The morphology and size of purified nanoparticles were analyzed using transmission emission microscopy (TEM, JEOL-2010, JEOL) operated at 200 kV. The particles were negatively-stained (1% uranyl acetate) prior to TEM analysis. Dynamic light scattering (DLS) and Zeta potential measurements were performed on Zetasizer Nanoseries (Nano-ZS, Malvern Instruments). The fluorescence measurements were performed at room temperature using a Hitachi F-4700 spectrofluorometer with a 200 W Xe-lamp as an excitation source.
F 4700 spectrofluorometer
Manufactured by Hitachi
The Hitachi F-4700 spectrofluorometer is a laboratory instrument used for the analysis of fluorescent samples. It measures the intensity of fluorescent light emitted by a sample when it is exposed to an excitation light source. The instrument is designed to provide accurate and reliable data on the fluorescent properties of a wide range of materials, including organic and inorganic compounds, biomolecules, and nanomaterials.
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3 protocols using f 4700 spectrofluorometer
1
Nanoparticle Characterization by TEM, DLS, Zeta
2
Chymotrypsin-AzC Interaction Probed by Fluorescence
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To understand the binding nature of chymotrypsin, fluorescence quenching experiments are used quite often. The fluorescence measurements were conducted on Hitachi F-4700 spectrofluorometer equipped with a temperature controller using 1.0 cm path length fluorescence cuvette. The Chymo molecule was taken as a fluorescent probe to study the interaction with AzC dye of different concentrations in the presence and absence of ZnONPs (1.0 mM). The excitation wavelength was taken at 295 nm to selectively excite the tryptophan unit at fixed slit width of 5.0 nm. The samples were prepared in phosphate buffer at pH 7.4 and the spectra were taken in the range of 300–450 nm wavelength at two different temperatures, 298 K and 323 K.
3
Spectroscopic Characterization of Organic Compounds
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1 H and 13 (link) C NMR spectra were recorded on a Bruker AV 400M spectrometer using chloroform-d solvent and tetramethylsilane as internal reference. J-values are given in Hz. High-resolution mass spectra (HRMS) were recorded on a LC/MS/MS, which consisted of a HPLC system (Ultimate 3000 RSLC, Thermo Scientific, USA) and a Q Exactive Orbitrap (QE orbitrap type) mass spectrometer. UV-vis absorption spectra and photoluminescence (PL) spectra were recorded on a Shimadzu UV-2600 and the Hitachi F-4700 spectrofluorometer. PL quantum yields were measured using absolute methods using a Hamamatsu C11347-11 Quantaurus-QY Analyzer. The lifetime was recorded on an Edinburgh FLS 980 instrument and measured using a time-correlated single-photon counting method. Thermogravimetric analysis was carried on a Mettler Toledo TGA/DSC3+ under dry nitrogen at a heating rate of 10 o C/min. The quantum chemistry calculation was performed on the Gaussian 09 (B3LYP/6-311G (d,p) basis set) software package.
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