Fluorescence excitation and emission peaks of the three fluorescence color morphs used for the experiment were determined from the collected colonies using the Varian Cary Eclipse Fluorescence Spectrometer (Agilent, USA) from host protein extracts.
Varian cary eclipse fluorescence spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The Varian Cary Eclipse Fluorescence Spectrometer is a laboratory instrument designed for the analysis of fluorescent samples. It is capable of measuring the fluorescence emission and excitation spectra of various materials.
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4 protocols using varian cary eclipse fluorescence spectrometer
1
Fluorescence Spectrometry of Protein
2
Critical Micelle Concentration Determination
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Critical Micelle Concentration (CMC) values of the synthetic compounds were determined by pyrene fluorescence method28 (link) using a Varian Cary Eclipse Fluorescence spectrometer (Agilent Technology, California, USA). The respective solutions were prepared using water previously saturated with pyrene (1 µM final concentration) in different concentrations (0.000001, 0.00001, 0.0001, 0.001, 0.01, 0.1, 1, 2.5, 5, 10, 15, 20 mM). Emission spectra of pyrene were obtained by exciting the samples at 334 nm. The fluorescence intensity ratio of I1/I3 (I1 = 373 nm, I3 = 383 nm) was plotted against sample solution concentration. The concentration at which the first break occurs was referred as the CMC value of the compound in water. Measurements were determined in triplicate.
3
Fluorescence Spectroscopy of Tetracaine
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Fluorescence emission spectra were recorded using a fluorescence spectrometer fitted with a Xenon pulse lamp (Varian Cary Eclipse Fluorescence Spectrometer, Agilent Technologies, UK). Beer-Lambert's law can only be applied over a limited range of optical densities and at a high sample optical density, attenuation due to absorption of the incident light or the emitted light can lead to a decrease in intensity and a possible change in spectral distribution [24, 25] . In light of the possibility of having a deviation in linearity due to the concentration of tetracaine and not the molecular aggregation, a Quartz fluorescence cells (Helima Fluorescence Cell, Helima UK Ltd., UK) with a 3 mm path length, with an off-centre illumination was used to record the measurements.
Excitation and emission slits were fixed at 5 nm. In all measurements, the excitation wavelength was set at 310 nm. The samples were scanned from 320 to 450 nm at a wavelength scan rate of 120 nm/min with a PMT detector gain of 600 V. The data were smoothed with a Savitzky Golay function filter size 25 using the Cary Eclipse software.
The experiments were performed at a temperature of 32 °C. The system was chemically stable over 6 days and the effect of ion pairing with the ions dissolved in the aqueous solution was not significant (Data not shown).
Excitation and emission slits were fixed at 5 nm. In all measurements, the excitation wavelength was set at 310 nm. The samples were scanned from 320 to 450 nm at a wavelength scan rate of 120 nm/min with a PMT detector gain of 600 V. The data were smoothed with a Savitzky Golay function filter size 25 using the Cary Eclipse software.
The experiments were performed at a temperature of 32 °C. The system was chemically stable over 6 days and the effect of ion pairing with the ions dissolved in the aqueous solution was not significant (Data not shown).
4
Characterization of Photoluminescent Quantum Dots
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QDs photoluminescent spectra were performed with the Varian Cary Eclipse Fluorescence Spectrometer (Agilent, Santa Clara, CA, USA) equipped with a xenon discharge lamp (peak power equivalent to 75 kW), a Czerny-Turner monochromator and photomultiplier tube detector (Model R-298). The emission spectra (1 nm data interval), which presented a maximum emission wavelength at 590 nm, were recorded upon excitation at 290 nm, with a delay time of 0.2 ms and a gate time of 5 ms. Excitation and emission slits were set at 10/10 nm respectively, and the averaging time selected to perform the experiments was 5 ms. A microplate reader accessory was used for phosphorescent immunoassay measurements.
QDs characterization was carried out by simultaneous detection and quantification of the elements constituting the QD core (S, Zn and Mn) using an ICP-MS/MS system (Agilent 8800 ICPQQQ, Tokyo, Japan). For separation and characterization of nanoparticles and their bioconjugates the ICP-MS/MS was coupled on-line to the AF4 system (AF2000, Postnova Analytics, Landsberg, Germany). Separation conditions are summarized in Table S1 (Electronic Supplementary Material, ESM). Dynamic light scattering spectra were measured by using a NanoZS90 instrument from Malvern Instruments, Houston, TX, USA.
QDs characterization was carried out by simultaneous detection and quantification of the elements constituting the QD core (S, Zn and Mn) using an ICP-MS/MS system (Agilent 8800 ICPQQQ, Tokyo, Japan). For separation and characterization of nanoparticles and their bioconjugates the ICP-MS/MS was coupled on-line to the AF4 system (AF2000, Postnova Analytics, Landsberg, Germany). Separation conditions are summarized in Table S1 (Electronic Supplementary Material, ESM). Dynamic light scattering spectra were measured by using a NanoZS90 instrument from Malvern Instruments, Houston, TX, USA.
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