The details of our experimental methods have been described previously[28 (link)]. Briefly, all CaAl2O4:Eu2+, Nd3+ powder samples were synthesized by a high-temperature solid-state reaction. High-purity SrCO3, Eu2O3 (Rhône-Poulenc, 99.99%), Nd2O3, Al2O3, MCO3 (M = Ca, Sr, and Ba; Merck, >99.0%), and SiO2 (Aerosil OX 50, Degussa) were mixed and H3BO3 was added as a flux. After grinding the mixtures in an agate mortar, they were fired in molybdenum crucibles for 3–5 h at ~1300°C in a furnace under a weak reductive atmosphere of flowing N2/H2 (5%) gas. After cooling down the synthesized samples to room temperature, they were ground again in an agate mortar. The final samples were irradiated with 365 nm UV-light for 5 min. After turning off the UV lamp, the emission spectra were recorded with a Hitachi 850 fluorescence spectrophotometer in the wavelength range from 300 to 950 nm.
850 fluorescence spectrophotometer
Manufactured by Hitachi
Sourced in Japan
The 850 Fluorescence Spectrophotometer is a laboratory instrument designed for the analysis of fluorescent samples. It is capable of measuring the emission and excitation spectra of fluorescent compounds, providing users with quantitative data on the fluorescent properties of their samples.
Automatically generated - may contain errors
Lab products found in correlation
4 methylumbelliferone 4 mu, by Merck Group (3 mentions)
Aerosil ox50, by Evonik (2 mentions)
D max 2500 pc, by Rigaku (1 mentions)
240 elemental analyzer, by PerkinElmer (1 mentions)
4 methylumbelliferyl β d glucuronide, by Merck Group (1 mentions)
D5200, by Nikon (1 mentions)
X gluc, by Merck Group (1 mentions)
Nicolet evolution 300 uv vis spectrophotometer, by Thermo Fisher Scientific (1 mentions)
11 protocols using 850 fluorescence spectrophotometer
1
Synthesis and Characterization of CaAl2O4:Eu2+,Nd3+ Phosphors
2
Synthesis and Characterization of Rare-Earth Phosphors
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The starting materials were high-purity SrCO3, Al2O3, Eu2O3 (Rhône-Poulenc, 99.99%), Dy2O3, MCO3 (M = Ca, Sr, Ba; Merck, > 99.0%), and SiO2 (Aerosil OX 50, Degussa). Small quantities of H3BO3 or B2O3 (0.1–0.3M) were added as a flux. The starting materials were weighed out in various amounts, mixed homogeneously, and ground in an agate mortar. Then, the dried powder mixtures were fired in molybdenum crucibles at ~1300°C for 3–5 h, under a weak reductive atmosphere of flowing N2-H2 (3%) gas, in horizontal tube furnaces. After a high-temperature solid-state reaction, the synthesized samples were cooled to room temperature in the furnace, and were ground again in an agate mortar. For the afterglow measurements, the samples were irradiated with 365 nm light for 5 min, and the emission spectra of the phosphors were recorded by a Hitachi 850 fluorescence spectrophotometer, from 300 to 950 nm. The decay curves of afterglow were measured with an ST-86LA brightness meter. All measurements were carried out at room temperature.
3
Thermal and Spectroscopic Characterization of Analytes
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All reagents were of analytical grade and obtained from commercial sources without further purification. Elemental analysis for C and H were performed on a PerkinElmer 240 elemental analyzer. Thermal analyses were performed on a SDT 2960 thermal analyzer from room temperature to 800 °C at a heating rate of 20 °C min−1 under nitrogen flow. Powder X-ray diffraction (PXRD) data were collected on a Rigaku D/Max-2500PC diffractometer with Cu Kα radiation (λ = 1.5406 Å) over the 2θ range of 5–50° with a scan speed of 5° min−1 at room temperature. Room temperature PL spectra and time-resolved lifetime were conducted on an Edinburgh FLS1000 fluorescence spectrometer equipped with a xenon arc lamp (Xe900), nanosecond flash-lamp (nF900) and a microsecond flashlamp with time-resolved single photon counting-multi-channel scaling (MCS) mode. Fluorescence spectra for the aqueous solutions of each analyte were recorded on a Hitachi 850 fluorescence spectrophotometer.
4
Determination of Selenium Content
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To determine the Se content of the feed, plasma, ovary, magnum, isthmus, and shell gland, all samples (0.5–1 g for feed, ovary, magnum, isthmus, and shell gland and 0.5–1 mL for plasma) were digested in a mixture of concentrated nitric acid and perchloric (2 HNO3:1 HClO4) for about 2 h at 200 °C until white fumes appeared. Then, 5 mL hydrochloric acid (1 HCl: 4 H2O) solution was added in the mixture and heated until white fumes appeared. After the mixture cooled, 20 mL EDTA was added in the digested sample and pH was adjusted to 1.5–2.0. After this, 3 mL 2,3 DiAminoNaphthalene (DAN) was added in the mixture and heated in the boiled water for 5 min. After the mixture cooled at room temperature, 4 mL cyclohexane was added and the mixture was shaken for 8 min. The supernatant was measured by fluorescence method using a Hitachi 850 fluorescence spectrophotometer (Tokyo, Japan) [16 (link)].
5
GUS Histochemical Localization and Quantification
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GUS histochemical location was conducted as previously described [1 (link)]. Each photograph in Fig. 2 f is representative of at least nine tissues from three replicated experiments. Fluorometric quantitative analysis was measured according to Jefferson’s method [16 (link)]. The control or transgenic samples were analysed to determine GUS activities with the substrate of 1 mM 4-methyl umbelliferyl β-d -glucuronide (Sigma-Aldrich, Shanghai, China). Fluorescence values were recorded with a Hitachi 850 Fluorescence spectrophotometer (Hitachi, Tokyo, Japan). The protein concentration was determined as described by Bradford [17 (link)].
6
Phosphor Blending via Solid-State Reaction
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For the chemical mixing
of two or three different phosphors via
high-temperature solid-state reaction, all starting materials were
first weighed, mixed homogeneously, and subsequently ground using
an agate mortar. The ground powder mixtures were fired in molybdenum
crucibles under a weak reductive atmosphere of N2–H2 gas at ∼1300 °C for 3–5 h. For chemical
mixing of two different phosphors by firing after high-temperature
solid-state reactions of each phosphor, the starting materials for
each phosphor were first weighed and subsequently mixed and ground
in an agate mortar separately. Afterward, each phosphor was synthesized
separately via a high-temperature solid-state reaction and the separately
synthesized phosphors were mixed at ratios of 10:0 or 5:5 in an agate
mortar and fired in the presence of H3BO3 at
800–1400 °C. The synthesized powders were cooled to room
temperature and ground again in an agate mortar. To measure the afterglow
intensity, the samples were irradiated at 365 nm for 5 min. After
irradiation, the emission spectrum of each sample was recorded using
a Hitachi 850 fluorescence spectrophotometer from 300 to 950 nm at
room temperature. The decay curves from the measured spectra were
fitted by three exponential components to calculate the lifetimes,
which are the inverse decay rates.
of two or three different phosphors via
high-temperature solid-state reaction, all starting materials were
first weighed, mixed homogeneously, and subsequently ground using
an agate mortar. The ground powder mixtures were fired in molybdenum
crucibles under a weak reductive atmosphere of N2–H2 gas at ∼1300 °C for 3–5 h. For chemical
mixing of two different phosphors by firing after high-temperature
solid-state reactions of each phosphor, the starting materials for
each phosphor were first weighed and subsequently mixed and ground
in an agate mortar separately. Afterward, each phosphor was synthesized
separately via a high-temperature solid-state reaction and the separately
synthesized phosphors were mixed at ratios of 10:0 or 5:5 in an agate
mortar and fired in the presence of H3BO3 at
800–1400 °C. The synthesized powders were cooled to room
temperature and ground again in an agate mortar. To measure the afterglow
intensity, the samples were irradiated at 365 nm for 5 min. After
irradiation, the emission spectrum of each sample was recorded using
a Hitachi 850 fluorescence spectrophotometer from 300 to 950 nm at
room temperature. The decay curves from the measured spectra were
fitted by three exponential components to calculate the lifetimes,
which are the inverse decay rates.
7
Histochemical Analysis of GUS Activity
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For the histochemical analysis of GUS, the infiltrated tobacco leaves were immersed in GUS staining solution (100 mM sodium phosphate buffer, pH 7.0, 0.5 mM K3Fe(CN)6, 0.5 mM K4Fe(CN)6, 10 mM Na2EDTA, 0.5 mg⋅mL-1 5-bromo-4-chloro-3-indolyl β-D -glucuronide (X-GluC), 0.1% Triton X-100) for 12 h at 37°C in darkness and were then detained in multiple changes of 75% ethanol. Further, GUS activity was expressed as nmol 4-methylumbelliferone (4-MU, Sigma-Aldrich) generated from the corresponding glucuronide (4-methylumbelliferyl β-D -glucuronide, MUG) per minute per milligram of soluble protein (Jefferson, 1987 (link)). To measure this activity, the 4-MU product was quantified by fluorescence intensity with a Hitachi 850 Fluorescence Spectrophotometer (Hitachi, Tokyo, Japan). The protein concentration was determined by the method of Bradford (Bradford, 1976 (link)).
8
N. benthamiana Transgenic Assay Protocol
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N. benthamiana leaves were used and sterile cultured for transgenic assay with EHA105 containing pCXGUS‐GUS‐P‐NbJAZ3. The GUS staining method was referred to the previous studies (Wu et al., 2020 (link); Zhang et al., 2019a (link)). Different tissues of the transgenic plants (young seedlings, mature leaves, roots, stems, flower petals, and calyxes) were collected and soaked in precooled 90% acetone solution for 30 min. Precooled GUS substrate containing 2 mM K4Fe(CN)6, 2 mM K3Fe(CN)6, 0.2% Triton‐X‐100, and 50 mM PBS (Na+, PH7.2) was used to gently flush all the tissues twice. All tissues were immersed into 100 mM X‐Gluc staining buffer for vacuum osmosis treatment for 15–20 min and transferred to a 37°C incubator for overnight incubation. Finally, all tissues treated by different ethanol concentrations (30, 50, 70, 90, and 100%) for decolorization were observed and photographed with the single‐lens reflex camera (Nikon D5200).
Leaves of the proNbCycB2‐GFP‐GUS transgenic tobacco line were used and sterile cultured for transgenic assay with EHA105 containing P35S:NbJAZ3 and P35S:NbWo. After 72 h injection, the leaves were stained with GUS substrate. The quantitative GUS assays were conducted with a Hitachi 850 Fluorescence spectrophotometer (Hitachi).
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N. benthamiana leaves were used and sterile cultured for transgenic assay with EHA105 containing pCXGUS‐GUS‐P‐NbJAZ3. The GUS staining method was referred to the previous studies (Wu et al., 2020 (link); Zhang et al., 2019a (link)). Different tissues of the transgenic plants (young seedlings, mature leaves, roots, stems, flower petals, and calyxes) were collected and soaked in precooled 90% acetone solution for 30 min. Precooled GUS substrate containing 2 mM K4Fe(CN)6, 2 mM K3Fe(CN)6, 0.2% Triton‐X‐100, and 50 mM PBS (Na+, PH7.2) was used to gently flush all the tissues twice. All tissues were immersed into 100 mM X‐Gluc staining buffer for vacuum osmosis treatment for 15–20 min and transferred to a 37°C incubator for overnight incubation. Finally, all tissues treated by different ethanol concentrations (30, 50, 70, 90, and 100%) for decolorization were observed and photographed with the single‐lens reflex camera (Nikon D5200).
Leaves of the proNbCycB2‐GFP‐GUS transgenic tobacco line were used and sterile cultured for transgenic assay with EHA105 containing P35S:NbJAZ3 and P35S:NbWo. After 72 h injection, the leaves were stained with GUS substrate. The quantitative GUS assays were conducted with a Hitachi 850 Fluorescence spectrophotometer (Hitachi).
9
Histochemical GUS Staining and Quantification
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For histochemical GUS staining, the infiltrated tobacco leaves were incubated in GUS staining solution with 50 mM sodium phosphate (pH 7.0), 10 mM Na2EDTA, 0.5 mM K4Fe(CN)6·3H2O, 0.1% Triton X-100 and 1 mM X-Gluc (Sigma-Aldrich, Shanghai, China) at 37 °C for 24 h and then cleared with 70% ethanol [90 (link)]. To monitor the activity of the DkLOX3 and CaMV35S promoters, quantitative GUS assays were performed according to the previously described method [90 (link)]. The fluorescence of the methylumbelliferone products was quantified with a Hitachi 850 fluorescence spectrophotometer (Hitachi, Tokyo, Japan). The total concentration of the protein extract from the tested samples was normalized using an established protocol [91 (link)]. GUS activity was expressed as nM of 4-methylumbelliferone (4-MU, Sigma-Aldrich) generated per minute per milligram of soluble protein. The GUS measurements were repeated at least three times with similar results.
10
Quantitative GUS Assay Protocol
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Histochemical and quantitative GUS assays were carried out according to the procedure published by Jefferson (1987 (link)). Histochemical staining of the leaves was performed as previously described (Xu et al., 2010 (link)). Quantitative GUS assays, using 4-methyl umbelliferyl glucuronide (MUG; Sigma-Aldrich) as a substrate, were quantified using a Hitachi 850 fluorescence spectrophotometer (Hitachi, Tokyo, Japan), and protein concentrations were measured using the protein–dye binding assay (Bradford, 1976 (link)) and a Nicolet Evolution 300 UV-VIS spectrophotometer (Thermo Electron Corp., Madison, WI, USA) with bovine serum albumin as the standard. GUS activity was expressed as nM of 4-methylumbelliferone (4-MU; Sigma-Aldrich) generated per minute per milligram of soluble protein. Data were subjected to one-way ANOVA for comparison of the means, and one-sided paired t-test were performed to assess significant differences between mock and treatment conditions using the SigmaPlot 10.0 statistical software package (Ashburn, VA, USA). Differences at P < 0.05 were considered significant.
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