Emission spectra were recorded on a Thermoscientific Lumina Fluorescence spectrometer, using a 150 W Xenon Lamp as the excitation source. Data were processed with Luminous software.
Lumina fluorescence spectrometer
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Lumina fluorescence spectrometer is a versatile instrument designed for the detection and analysis of fluorescent samples. It measures the emission spectra of fluorescent compounds and provides quantitative data on their intensity and wavelength characteristics.
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Lab products found in correlation
Peltier temperature controller, by Thermo Fisher Scientific (2 mentions)
Mitosox red, by Thermo Fisher Scientific (2 mentions)
X pert mpd diffractometer, by Malvern Panalytical (2 mentions)
Cary 500 scan uv vis nir spectrophotometer, by Agilent Technologies (1 mentions)
Uv 3600, by Shimadzu (1 mentions)
N n diisopropylethylamine dipea, by Merck Group (1 mentions)
A300 10 12, by Bruker (1 mentions)
Maldi 7090, by Shimadzu (1 mentions)
Enhanced hrp dab chromogenic substrate kit, by Tiangen Biotech (1 mentions)
D8 advance x ray powder diffractometer, by Bruker (1 mentions)
Escalab 250xi x ray photoelectron spectroscopy, by Thermo Fisher Scientific (1 mentions)
Pb 10 ph meter, by Sartorius (1 mentions)
Uv 3600 plus uv vis nir spectrophotometer, by Shimadzu (1 mentions)
Thermo nicolet is10 spectrometer, by Thermo Fisher Scientific (1 mentions)
Chi 760e, by Chenhua (1 mentions)
Su8010, by Hitachi (1 mentions)
Jem 1011, by JEOL (1 mentions)
U 4100, by Hitachi (1 mentions)
Super dynamic iiwv cp460, by Panasonic (1 mentions)
Flsp920, by Edinburgh Instruments (1 mentions)
38 protocols using lumina fluorescence spectrometer
1
Fluorescence Spectra Acquisition
2
Fluorescence and Absorbance Spectroscopy
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Fluorescence spectra
were recorded with a LUMINA fluorescence spectrometer (Thermo, USA)
at room temperature. Absorbance measurements were performed on a Cary
500 Scan UV/vis/NIR spectrophotometer (Varian, USA).
were recorded with a LUMINA fluorescence spectrometer (Thermo, USA)
at room temperature. Absorbance measurements were performed on a Cary
500 Scan UV/vis/NIR spectrophotometer (Varian, USA).
3
Synthesis and Characterization of NIR Emitting Compounds
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2,3,3-Trimethylindolenine, 1,4-Butanesulfone, 4-Hydroxyphenyl boronic acid and 4-Amine-2,2,6,6-tetramethylpiperidinyloxy (4-amine-TEMPO) were purchased from Adamas; Tetrakis (triphenylphosphine) palladium, N,N-Diisopropylethylamine (DIPEA) and anhydrous ethanol were purchased from Sigma. All reactions were monitored by thin-layer chromatography (TLC). Flash chromatography was carried out using silica gel (200–300 mesh). 1H NMR spectra were recorded with an Agilent 600 MHz with trimethylchlorosilane (TMS) as an internal standard. Mass spectrometry (HRMS) was performed on a Bruker Ultraflextreme MALDI-TOF system or Shimadzu MALDI-7090. Electron paramagnetic resonance (EPR) was tested with Bruker A300-10/12. NIR spectroscopy was performed using a NIR spectrophotometer (Shimadzu, UV-3600). Photoluminescence was determined with a NIR fluorescence spectrometer (Lumina Fluorescence Spectrometer, Thermo Fisher, Waltham, MA, USA).
4
Quantitative Analysis of Recombinant mCherry in E. coli
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The fluorescent signal of mCherry produced in recombinant E. coli was measured with the Lumina fluorescence spectrometer (Thermo, USA) as previously described [22 (link)]. Fluorescence emission was recorded at 610 nm for mCherry, and the fluorescence value was normalized by dividing the fluorescence intensity by the OD600 value of the same sample. A dot blot using cell lysate was performed as previously described [29 (link)]. Briefly, twenty microliters of induced culture was mixed with an equal volume of assay solution (200 mM Tris-HCl, pH 6.8, 2% SDS, and 200 mM DTT), and boiled for 10 min. An aliquot of 5 μL of each sample was dripped onto PVDF membrane. After the membrane was blocked and washed, it was incubated with primary mouse anti-His IgG (Tiangen, Beijing, China). After extensive washing, the membrane was then incubated with a secondary HRP labeled rabbit anti-mouse IgG (Sangon Biotech, Shanghai, China). Finally, the associated antibodies were visualized using an enhanced HRP-DAB chromogenic substrate kit (Tiangen, Beijing, China).
5
Fluorescence Spectroscopy of Protein Samples
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The fluorescence spectra of HIU-treated and untreated protein samples were obtained by a Lumina fluorescence spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). The protein samples were dissolved in phosphate buffer (10 mM, pH 7.0) to obtain a concentration of 0.2 mg/mL. The protein solution was excited at a wavelength of 290 nm and the fluorescence emission was recorded from between 300 and 500 nm at a bandwidth of 5 nm for both excitation and emission.
6
Quantifying Fluorescent Biosensor Response
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Both of the eGFP and mCherry fluorescent proteins generated from three whole-cell biosensors with Cd(II) exposure were quantitated with a Lumina Fluorescence Spectrometer (Thermo Fisher Scientific, Waltham, MA, United States) as previously described (Hui et al., 2019 (link)). After the cultures were properly diluted in purified H2O, 3 ml of sample was added to a low fluorescence background quartz cuvette. The excitation wavelength was set at 488 nm, and the intensity of emitted eGFP fluorescence was recorded at 507 nm. The excitation wavelength was set at 587 nm, and the intensity of emitted mCherry fluorescence was recorded at 610 nm. The fluorescent signal is indicated as a fluorescence count value (unit = cnt), and normalized by dividing by the OD600 value of the same culture (Hui et al., 2018c ).
7
Quantifying mCherry Fluorescence in E. coli
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The fluorescence intensity of mCherry produced in E. coli was measured as previously described (Hui et al., 2018c (link)). Briefly, E. coli cells were collected, washed, and resuspended in 50 mM PBS. A 3-mL aliquot of E. coli suspension or diluent was added to 1-cm low fluorescence background quartz cuvette. To test the fluorescence intensity of induced mCherry, the excitation wavelength was set at 587 nm and the intensity of emitted fluorescence of mCherry at 610 nm was recorded with Lumina fluorescence spectrometer (Thermo Fisher Scientific, United States). The fluorescence intensity was normalized by dividing the fluorescence intensity at the emission wavelength 610 nm by the OD600 value of the same sample. The background value was obtained from the control assays with uninduced cell suspensions.
8
UV-Vis and Fluorescence Spectroscopy
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Absorption spectra in the 250–750 nm range were recorded on a Biochrom WPA 80-3003-75 Model Biowave II UV/Visible Life Science Spectrophotometer. Emission spectra were recorded on a Thermoscientific Lumina Fluorescence spectrometer, using a 150 W Xenon Lamp and the excitation source set to the absorbance maximum of the compound in the 300–700 nm range. The spectral bandwidth was 5 nm, with a 2 second integration time. Data were acquired with Luminous software.
9
Advanced Material Characterization Techniques
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TEM and HRTEM images were taken with
a JEM-2100F high-resolution transmission emission microscope (JEOL,
Japan). XRD spectrum was measured on a Bruker D-8 Advance Powder X-ray
diffractometer (Bruker, Germany). XPS spectra were taken with ESCALAB
250Xi X-ray photoelectron spectroscopy (Thermo Fisher Scientific).
FT-IR spectrum was recorded on a Thermo Nicolet iS10 spectrometer
(Thermo Fisher Scientific). Elemental analysis was carried out on
a Vario EL/Micro Cube organic element analyzer (Elementar Analysensysteme
GmbH, Germany). Magnetic property was recorded by using a VersaLab
Vibration Sample Magnetometer (Quantum Design). UV–vis absorption
spectra were recorded on a Shimadzu UV-3600 Plus UV–vis–NIR
spectrophotometer (Shimadzu, Japan). Fluorescence spectra were recorded
on a Thermo Scientific Lumina fluorescence spectrometer (Thermo Fisher
Scientific). Time-resolved fluorescence spectra were measured on a
Horiba Scientific QM-8075 high sensitivity steady-state transient
fluorescence spectrometer (HORIBA, Japan). ζ-Potential was recorded
on a Zetasizer Nano ZS (Malvern, U.K.). CV curves were obtained from
a Chenhua CHI-760E electrochemical workstation (Shanghai, China).
The pH values were mediated using a Sartorius PB-10 pH meter (Sartorius,
China).
a JEM-2100F high-resolution transmission emission microscope (JEOL,
Japan). XRD spectrum was measured on a Bruker D-8 Advance Powder X-ray
diffractometer (Bruker, Germany). XPS spectra were taken with ESCALAB
250Xi X-ray photoelectron spectroscopy (Thermo Fisher Scientific).
FT-IR spectrum was recorded on a Thermo Nicolet iS10 spectrometer
(Thermo Fisher Scientific). Elemental analysis was carried out on
a Vario EL/Micro Cube organic element analyzer (Elementar Analysensysteme
GmbH, Germany). Magnetic property was recorded by using a VersaLab
Vibration Sample Magnetometer (Quantum Design). UV–vis absorption
spectra were recorded on a Shimadzu UV-3600 Plus UV–vis–NIR
spectrophotometer (Shimadzu, Japan). Fluorescence spectra were recorded
on a Thermo Scientific Lumina fluorescence spectrometer (Thermo Fisher
Scientific). Time-resolved fluorescence spectra were measured on a
Horiba Scientific QM-8075 high sensitivity steady-state transient
fluorescence spectrometer (HORIBA, Japan). ζ-Potential was recorded
on a Zetasizer Nano ZS (Malvern, U.K.). CV curves were obtained from
a Chenhua CHI-760E electrochemical workstation (Shanghai, China).
The pH values were mediated using a Sartorius PB-10 pH meter (Sartorius,
China).
10
Fluorescence-Based Enzyme Kinetics Assay
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The enzymatic reaction was carried out using a Lumina fluorescence spectrometer (Thermo Fischer Scientific Inc.) coupled with a Peltier system 4-Position Cell Holder Fluorescence device to control the agitation speed and assay temperature. Reactions with Abz-KLRSSKQ-EDDnp substrate were carried out in a quartz cuvette with an optical path length of 10 mm. The wavelengths were set to λex: 320 nm and λem: 420 nm. Data were collected and analyzed using Luminous software version 3.0.
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