Fluorescence microplate reader

Manufactured by Molecular Devices

Sourced in United States

The Fluorescence microplate reader is a laboratory instrument used to detect and quantify fluorescent signals in multi-well microplates. It measures the intensity of fluorescence emitted by samples in each well of the microplate, providing data on the fluorescent properties of the samples.

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Lab products found in correlation

H2dcfda, by Thermo Fisher Scientific (4 mentions) Dcfh da, by Merck Group (3 mentions) Cyquant cell proliferation assay, by Thermo Fisher Scientific (3 mentions) Amplex red hydrogen peroxide assay kit, by Thermo Fisher Scientific (3 mentions) Fluorescence microscope, by Zeiss (2 mentions) Fluorescence microscope, by Olympus (2 mentions) Dcfh da, by Beyotime (2 mentions) 6 well collagen coated plate, by BD (2 mentions) Bodipy tr cadaverine bc, by Thermo Fisher Scientific (2 mentions) In situ cell death detection kit, by Roche (1 mentions) Matrigel, by BD (1 mentions) Calcein am, by Dojindo Laboratories (1 mentions) Odyssey imaging system, by LI COR (1 mentions) Dcf da solution, by Merck Group (1 mentions) Vet scan analyzer, by Abaxis (1 mentions) Lsm 710 meta confocal microscope, by Zeiss (1 mentions) Daf 2da, by Merck Group (1 mentions) Dcfh da, by Corning (1 mentions) Cyquant, by Thermo Fisher Scientific (1 mentions) Ponatinib, by Selleck Chemicals (1 mentions)

Spelling variants of this product

Microplate reader (2507 citations) Gemini EM fluorescence microplate reader (11 citations) Gemini XPS Fluorescence Microplate Reader (6 citations) Spectra Max M3 Fluorescence Microplate Reader (5 citations) Multiplate microplate fluorescence reader (4 citations) SpectraMax fluorescence microplate reader (3 citations) SpectraMax Gemini XPS Fluorescence Microplate Reader (2 citations) Fluorescence microplate reader for endpoint reading (2 citations) Spectramax M2/e fluorescence microplate reader (2 citations) Fluorescence-detecting microplate reader (2 citations)

99 protocols using fluorescence microplate reader

Apoptosis Quantification in Palmitate-Treated Cells

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For the TUNEL assay, TRB3 cells were seeded in 96-well plates (5×10⁴ cells per well) and treated with or without 500 ng/ml Dox for 48 h. The cells were then treated with 0.2 mM palmitate/BSA complex or BSA (control) in RPMI 1640 containing 10% FCS for 24 h. In some experiments, the cells were pretreated with or without rottlerin (5 µM Calbiochem, Nottingham, UK) for 1 h or Gö6850 (1 µM Calbiochem, Nottingham, UK) for 1 h before being treated with palmitate. The cells were then washed twice, fixed in 4% paraformaldehyde for one hour, and then permeabilized with 0.1% Triton X-100 in PBS/BSA solution. The TUNEL assay was performed using in situ cell death detection kits (Roche, Indianapolis, IN, USA), according to the manufacturer's protocols.
Caspase-3 activity was measured by caspase-3 assay kit (Molecular Probes, Invitrogen, Carlsbad, CA, USA) following the manufacturer's instruction on harvested cells treated as indicated. Microplate fluorescence reader (Molecular Devices Corp, Sunnyvale, CA, USA) was used for detection.

Qin J., Fang N., Lou J., Zhang W., Xu S., Liu H., Fang Q., Wang Z., Liu J., Men X., Peng L, & Chen L. (2014). TRB3 Is Involved in Free Fatty Acid-Induced INS-1-Derived Cell Apoptosis via the Protein Kinase C δ Pathway. PLoS ONE, 9(5), e96089.

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Matrigel-coated Transwell Invasion Assay

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FluoroBlok Transwell insert of 8-μm pore size (Corning) were coated by adding 30 ul diluted Matrigel (BD Biosciences; 9.7 mg/ml) and air-dried overnight. The coated inserts were placed on a 24-well plate containing DMEM supplemented with 10% FBS as a chemo-attractant. The cells (2.5×10⁴) were suspended in DMEM containing 0.1% FBS and plated onto the insert. After 16-h incubation, the cells invaded to the lower side of the membrane were stained with Calcein-AM (Dojindo, Japan) for 10 min, and then recorded using Microplate Fluorescence Reader (Molecular Devices, USA).

Shi G., Yoshida Y., Yuki K., Nishimura T., Kawata Y., Kawashima M., Iwaisako K., Yoshikawa K., Kurebayashi J., Toi M, & Noda M. (2016). Pattern of RECK CpG methylation as a potential marker for predicting breast cancer prognosis and drug-sensitivity. Oncotarget, 7(50), 82158-82169.

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Monitoring SOS Response in E. coli

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SOS expression was monitored in log phase cells grown in LB medium at 37°C.[10 (link)] All strains contained plasmid P_sulA-gfp. Fluorescence was determined by measurement of emission at 525 nm with excitation at 470 nm in a microplate fluorescence reader (Molecular Device, CA, USA). Fluorescence intensity was expressed as arbitrary units per 10⁹ cells. SOS response induction by UV was carried out using the UV oven (Bio-Link, Vilber Lourmat, France) equipped with five fluorescent lamps of 8 W each, emitting from 180 to 280 nm with a peak at 254 nm. UV doses were programmed and are controlled by a radiometer that constantly monitors the UV light emission. For measuring induction of promoters by UV, 1-ml portions of lat-exponential-phase cultures (OD₆₀₀ = 0.6) of strains containing P_sulA-gfp fusion were irradiated for 2 min in an open Petri dish placed 25 cm beneath the lamp (the UV oven, 0.2 kJ/m²). Afterward, the cultures were used for measurement of production of gfp.

Huo Y.J., Qiao L., Zheng X.W., Cui C., Ma Y.F, & Lu F. (2015). MreBCD-associated Cytoskeleton is Required for Proper Segregation of the Chromosomal Terminus during the Division Cycle of Escherichia Coli. Chinese Medical Journal, 128(9), 1209-1214.

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Kinetic Aggregation of Tau Constructs

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Aggregation was induced by incubating BPS vesicles with K18 or the mimetic constructs at a molar ratio of 1:20 (protein:phospholipid) in 10 mM Hepes, pH 7.4, 100 mM NaCl, and 2.5 mM DTT at 37 °C. ThT was added immediately prior to shaking at 1000 rpm. The data were collected in triplicates at time intervals (0, 6, 12, 24, 30, 36, 48, and 72 h) using a microplate fluorescence reader (Molecular Devices). The excitation and emission wavelengths were 440 and 490 nm, respectively.
Half time to maximal fluorescence (T₅₀) of kinetic aggregation data was used as a measure of seeding competence for each tau construct. T₅₀ was determined by fitting aggregation data to Equation 3 (86 (link)), where y is the fluorescence intensity; A2 and A1 are the initial and maximum intensity signals, respectively; T is the measurement time, and dT is the time constant. Data were graphed and analyzed using GraphPad Prism version 8.0 for Mac OS X, GraphPad Software, www.graphpad.com.

Equation4.

Acosta D.M., Mancinelli C., Bracken C, & Eliezer D. (2021). Post-translational modifications within tau paired helical filament nucleating motifs perturb microtubule interactions and oligomer formation. The Journal of Biological Chemistry, 298(1), 101442.

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Characterization of PPA-JM-Phage Conjugate

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The ultraviolet-visible absorption spectra of PPA-JM-phage and JM-phage were recorded on a spectrometer (Molecular Devices). Fluorescence spectra of PPA-JM-phage with different ratios of PPA to JM-phage were obtained using a microplate fluorescence reader (Molecular Devices) with excitation of 326 nm and 671 nm. In addition, to confirm that PPA was covalently attached to the JM-phages, PPA-JM-phages were subjected to sodium dodecyl sulfate (SDS) polyacrylamide-gel electrophoresis (PAGE) and imaged using an Odyssey imaging system (LI-COR Biosciences, Lincoln, NE, USA). Details can be found in the Supplementary materials. The morphology of the PPA-JM-phage was analyzed by transmission electron microscopy (TEM; JEOL, Tokyo, Japan). Samples were prepared by depositing one drop of dilute solution onto a dry copper grid that had been coated with carbon in advance. Subsequently, the grid was stained with 2% uranium acetate before TEM observation.

Dong S., Shi H., Zhang X., Chen X., Cao D., Mao C., Gao X, & Wang L. (2018). Difunctional bacteriophage conjugated with photosensitizers for Candida albicans-targeting photodynamic inactivation. International Journal of Nanomedicine, 13, 2199-2216.

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Quantifying Intracellular Lipid Accumulation

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Intracellular lipid accumulation was quantified using Nile Red, a fluorescent dye that binds to neutral lipids. Cells were seeded in 96 well plates and incubated with chemicals for 24 h. On the following day, cells were fixed with 4% paraformaldehyde and stained with 1 μg/ml Nile Red solution. The fluorescence was measured with a microplate fluorescence reader (Molecular Devices) at excitation and emission wave lengths of 488 nm and 580 nm, respectively.

Lee W., Kim H.Y., Choi Y.J., Jung S.H., Nam Y.A., Zhang Y., Yun S.H., Chang T.S, & Lee B.H. (2022). SNX10-mediated degradation of LAMP2A by NSAIDs inhibits chaperone-mediated autophagy and induces hepatic lipid accumulation. Theranostics, 12(5), 2351-2369.

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Probiotic CMs Mitigate UV-induced ROS

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The protective effect of the probiotic CMs against UV-induced intracellular ROS production in NHDFs was assessed using a 2′,7′-dichlorodihydrofluorescein diacetate (DCF-DA, Sigma-Aldrich, USA) staining assay. Briefly, UVB-irradiated cells were treated with probiotic CMs for 24 h. The cells were then mixed with 10 μM DCF-DA solution (Sigma-Aldrich) and incubated for 1 h. The fluorescence intensity was determined using a microplate fluorescence reader (Molecular Devices, USA) with excitation wavelength at 485 nm and emission wavelength at 530 nm.

Hong Y.K., An S., Lee Y.H., Yang S.A., Yoon Y.K., Lee J., Lee G., Chung M.J, & Bae S. (2022). Potential anti-ageing effects of probiotic-derived conditioned media on human skin cells. Acta pharmaceutica (Zagreb, Croatia), 72(3).

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Quantifying Oxidative Stress in HepG2 Cells

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DCFH-DA is a cell-permeable, non-fluorescent probe that is cleaved by intracellular esterases and converted into highly fluorescent dichlorofluorescein upon reaction with H₂O₂. After 500 µM tert-butyl hydroperoxide (t-BHP) treatment with HepG2 cells for 3 h, the cells were stained with 10 µM DCFH-DA for 30 min at 37 °C. H₂O₂ generation was determined by measuring dichlorofluorescein using a fluorescence microscope (Zeiss, Germany) or fluorescence microplate reader (Jemini, Molecular Device) at excitation/emission wavelengths of 485/530 nm.

Kim J.M., Choi M.H, & Yang J.H. (2023). Cinnamomum japonicum Siebold Branch Extracts Attenuate NO and ROS Production via the Inhibition of p38 and JNK Phosphorylation. Molecules, 28(4), 1974.

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Measuring Serum AGEs in Animal Studies

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The serum samples obtained from the blood of the experimental animals were stored in sterile tubes at −80 °C until analysis. Blood urea nitrogen (BUN) and serum creatinine (SCr) concentrations were analyzed using a Vet Scan analyzer (Abaxis, Inc., Union City, CA, USA).
The OxiSelectTM AGEs Competitive ELISA Kit from Cell Biolabs (San Diego, CA, USA) was used to determine the concentrations of AGEs in the serum. The circulating AGEs were analyzed depending on the sample’s fluorescence or absorbance in each well, as measured using a fluorescence microplate reader (Molecular Devices, San Jose, CA, USA) at 450 nm.

Kundu A., Gali S., Sharma S., Park J.H., Kyung S.Y., Kacew S., Kim I.S., Lee K.Y, & Kim H.S. (2022). Tenovin-1 Ameliorates Renal Fibrosis in High-Fat-Diet-Induced Diabetic Nephropathy via Antioxidant and Anti-Inflammatory Pathways. Antioxidants, 11(9), 1812.

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Measuring Cellular ROS and NO Levels

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The cellular ROS and NO were measured by using 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) (Sigma-Aldrich Corp.) and 4,5-diaminofluorescein diacetate (DAF-2DA) (Sigma-Aldrich Corp.), respectively. 661W cells in a Corning Costar 96-well black clear-bottom plate at a density of 3 × 10³ cells/well were incubated with 1.8 μM atRAL in the presence or absence of IRBP for 2 hours, and then with 20 μM DCFH-DA or 10 μM DAF-2DA for 20 minutes. After washing with PBS twice, the relative fluorescence unit was measured at an excitation wavelength of 485 nm and an emission wavelength of 528 nm with a fluorescence microplate reader (Molecular Device). The cells incubated in the same condition were used for capturing fluorescent signals with a Zeiss LSM710 Meta confocal microscope with an ×20 objective lens. Each image was collected under the same system settings (e.g., the same laser intensity and scan speed).

Lee M., Li S., Sato K, & Jin M. (2016). Interphotoreceptor Retinoid-Binding Protein Mitigates Cellular Oxidative Stress and Mitochondrial Dysfunction Induced by All-trans-Retinal. Investigative Ophthalmology & Visual Science, 57(4), 1553-1562.

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