Varioskan flash spectrophotometer

Manufactured by Thermo Fisher Scientific

Sourced in United States, Spain, Finland, Italy

The Varioskan Flash spectrophotometer is a high-performance instrument designed for accurate and reliable absorbance measurements across a wide range of applications. It features a xenon flash lamp, which provides a broad spectrum of light for efficient detection. The Varioskan Flash supports multiple microplate formats and offers a wavelength range of 200 to 1000 nm, allowing for a variety of spectroscopic analyses.

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Lab products found in correlation

Cell proliferation kit 1, by Roche (3 mentions) Nicolet is50, by Thermo Fisher Scientific (3 mentions) Cytoselect ldh cytotoxicity assay kit, by Cell Biolabs (2 mentions) 96 well plate, by Corning (2 mentions) Cfx connect real time system, by Bio-Rad (2 mentions) Eclipse ti, by Nikon (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Cell counting kit 8 (cck8), by Immunoway (1 mentions) Milli q system, by Merck Group (1 mentions) Phloroglucinol, by Merck Group (1 mentions) Primescipt rt reagent kit, by Takara Bio (1 mentions) Sybr green primescript rt kit, by Takara Bio (1 mentions) Rnaiso kit, by Takara Bio (1 mentions) Tg dta system, by PerkinElmer (1 mentions) Zetasizer nano z, by Malvern Panalytical (1 mentions) Jem 2100 plus, by JEOL (1 mentions) Ph1100l, by Avantor (1 mentions) Cell counting kit, by Dojindo Laboratories (1 mentions) Eclipse ti microscope, by Nikon (1 mentions) Cck 8 kit, by BestBio (1 mentions)

48 protocols using varioskan flash spectrophotometer

UV-Vis Absorbance Calibration Curves for Antioxidants

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UV-Vis absorbance calibration curves were determined for gallic acid and ferulic acid at the conditions of the to-be-used extractive media (pH = 7.5,

P

= 0.1 MPa and

T

= 298.15 K) by measuring the UV-Vis absorbance of known concentrations of the antioxidants at the wavelength of local maxima (260 and 310 nm, respectively) using the Thermo Scientific Varioskan Flash spectrophotometer. These solutions were prepared by weighing the solutes and purified water in the ADAM AAA 250 L balance and pH was adjusted with a 0.5 M aqueous solution of NaOH. The pH measurements were performed with a Crison pH meter Basic 20. Next, the absorbances of the blanks (purified water and plate) were subtracted from the experimental values and a first-degree fitting was performed, obtaining the absorbance-concentration calibration curves, as Equation (5) shows.

A = α C + β

where

A

is absorbance,

α

is the slope of the calibration curve (absorptivity),

C

is the antioxidant concentration (in g·mL⁻¹) and

β

is the y-intercept.

Velho P., Rebelo C.S, & Macedo E.A. (2023). Extraction of Gallic Acid and Ferulic Acid for Application in Hair Supplements. Molecules, 28(5), 2369.

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Anticholinesterase Activity Evaluation

Cited 1 time

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Compounds 2a–2l were evaluated for anticholinesterase activities by the Ellman’s method [36 (link)] with slight modification as described in previous investigations [22 (link),37 (link),38 (link)]. The absorbances were determined spectrometrically at 412 nm on a Varioskan flash spectrophotometer (Thermo Scientific, Waltham, MA, USA). The IC₅₀ and standard deviation values were determined graphically using Graph Pad Prism.

Agbo E.N., Gildenhuys S, & Mphahlele M.J. (2019). Inhibitory Effects of Novel 7-Substituted 6-iodo-3-O-Flavonol Glycosides against Cholinesterases and β-secretase Activities, and Evaluation for Potential Antioxidant Properties. Molecules, 24(19), 3500.

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Cell Proliferation Monitoring Using CCK-8

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The Cell Counting Kit-8 (CCK-8, ImmunoWay Biotechnology Company, Plano, TX, USA) assay was used to monitor cell proliferation. In brief, the cells transfected with siRNA or plasmid were placed on 96-well plates and cultured for 24 h, 48 h, 72 h, and 96 h. Then, the OD450 value was detected using the Thermo Scientific Varioskan Flash spectrophotometer (Thermo Scientific, Finland).

Tang P., Wu Y., Zhu C., Li Q, & Liu S. (2023). Microdissecting the Hypoxia Landscape in Colon Cancer Reveals Three Distinct Subtypes and Their Potential Mechanism to Facilitate the Development of Cancer. Journal of Oncology, 2023, 9346621.

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Cytotoxicity and Metabolic Assays

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To assess cytotoxicity and metabolic turnover, we used Lactate dehydrogenase (LDH) assay (i.e., cytotoxicity) or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (i.e., metabolic turnover) as previously described [28 (link),37 (link)], with minor modifications. Cells were distributed in 96-well plates (Costar, Milan, Italy) at a final density of 10,000 cells/well/100 µL and incubated for 24 h. The day after, cells were exposed to drugs as above described, and incubated for 4, 24 and 48 h. On the day of each time point, medium was removed and processed as manufacturer’s instructions for the LDH-viability assay (CytoSelectTM LDH cytotoxicity assay kit, Cell Biolabs, Milan, Italy). For metabolic turnover, MTT at a final concentration of 1 mg/mL was added to each well and incubated for 3 h under standard culture conditions. Media were then gently removed, 200 µL of MTT solvent (DMSO) was added, and cells were stirred on an orbital shaker for 10 min at room temperature. The absorbance was measured using a Varioskan Flash spectrophotometer (Thermo Scientific, Milan, Italy) at 570 nm. Data were expressed as the percentage of MTT reduction versus control cells. Each experiment was performed three times with six replicates per condition during each experimental run.

Torrisi F., Alberghina C., Lo Furno D., Zappalà A., Valable S., Li Volti G., Tibullo D., Vicario N, & Parenti R. (2021). Connexin 43 and Sonic Hedgehog Pathway Interplay in Glioblastoma Cell Proliferation and Migration. Biology, 10(8), 767.

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Quantifying Cell Death by ELISA

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Cell death was assessed by ELISA according to the instructions of the manufacturer (Sigma-Aldrich 115446775001). Optical density at 405 nm against 490 nm reference was determined with a Varioskan Flash spectrophotometer (Thermo Scientific). In the case of primary hepatocytes, cells were seeded in 12-well plates. In the case of pancreatic islets, 10 islets per well were used.

Sola-García A., Cáliz-Molina M.Á., Espadas I., Petr M., Panadero-Morón C., González-Morán D., Martín-Vázquez M.E., Narbona-Pérez Á.J., López-Noriega L., Martínez-Corrales G., López-Fernández-Sobrino R., Carmona-Marin L.M., Martínez-Force E., Yanes O., Vinaixa M., López-López D., Reyes J.C., Dopazo J., Martín F., Gauthier B.R., Scheibye-Knudsen M., Capilla-González V, & Martín-Montalvo A. (2023). Metabolic reprogramming by Acly inhibition using SB-204990 alters glucoregulation and modulates molecular mechanisms associated with aging. Communications Biology, 6, 250.

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Seaweed Total Phenolic Content Quantification

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Total phenolic content (TPC) was measured by the Folin–Ciocalteu method adapted to microtiter plate measurement [84 (link)]. An aliquot (100 μL) of the extract was placed into a 2 mL conical tub and mixed with 500 μL of the Folin–Ciocalteu reagent, diluted 1:10 (Sigma-Aldrich, San Luis, MO, USA) with water (Milli-Q system, Millipore, Burlington, MA, USA), and 400 μL of 12.5% sodium carbonate (Sigma-Aldrich, San Luis, MO, USA) solution. The mixture was allowed to stand at 45 °C for 15 min. Samples were centrifuged at 5500 rpm for 5 min to remove the excess of sodium carbonate. An amount of 250 μL was transferred into a microtiter plate and the absorbance was read at 765 nm in a Varioskan Flash spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). A calibration curve was obtained using phloroglucinol (Sigma-Aldrich, San Luis, MO, USA) as standard and the range of concentrations used was 10–80 ng/μL. Each standard and sample solution were run in duplicate and results were expressed as milligrams of phloroglucinol equivalents (PGE) per gram of seaweed dry weight (mg PGE/g dw). The limits of detection (LOD) and quantification (LOQ) were calculated according to Long & Winefordner (1983) [85 (link)], assuming k values of 3 and 10, respectively.

Rubiño S., Peteiro C., Aymerich T, & Hortós M. (2022). Brown Macroalgae (Phaeophyceae): A Valuable Reservoir of Antimicrobial Compounds on Northern Coast of Spain. Marine Drugs, 20(12), 775.

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MTT Assay for Cell Viability

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MTT activity was determined using the Cell Proliferation Kit I according to the recommendations of the manufacturer (Roche, Spain). Optical density was determined at 575 nm with a reference wavelength of 690 nm using a Varioskan Flash spectrophotometer (Thermo Scientific, Spain). In the case of primary hepatocytes, cells were seeded in 12-well or 24-well plates. In the case of pancreatic islets, 35 islets per well were used.

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Profiling miRNA Expression in AEC-II Cells

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The miR-21-5p, miR-34c-5p, miR-449a-5p and U6 primers were designed by Boheng Biomart (Beijing, China). Total RNA from AEC-II cells was isolated using the RNAiso Kit (Takara Biotechnology Co., Ltd., Dalian, China). RNA concentration and purity was checked using a Varioskan Flash spectrophotometer (Thermo Fisher Scientific, Inc.). A total of 500 ng RNA was reverse transcribed to cDNA using the PrimeScipt RT Reagent Kit (Takara Biotechnology Co., Ltd.) in a final volume of 10 µl, according to the manufacturer's protocol. RT-PCR was performed with a CFX Connect Real-Time system (Bio-Rad Laboratories, Inc., Hercules, CA, USA) using a SYBR green PrimeScript RT kit (Takara Biotechnology Co., Ltd.). and the following thermocycling conditions: Initial denaturation at 95°C for 5 min, followed by 40 cycles of denaturation at 95°C for 10 sec and combined annealing/extension at 65.7°C for 30 sec. The primer sequences were as follows: miR-34c-5p forward, 5′-CCAGGCAGTGTAGTTAGCT-3′ and reverse, 5′-GTTGGCTCTGGTGCAGGGTCCGAGGTATTCGCACCAGAGCCAACGCAATC-3′; miR-449a-5p forward, 5′-AACGTGGCAGTGTATTGTTA-3′ and reverse, 5′-GTTGGCTCTGGTGCAGGGTCCGAGGTATTCGCACCAGAGCCAACACCAGC-3′; U6 forward, 5′-CTCGCTTCGGCAGCACACG-3′ and reverse, 5′-AACGCTTCACGAATTTGCGT-3′. U6 was used as an internal control. The relative expression levels were calculated using the 2^−ΔΔCq method (21 (link)).

Qin S., Chen M., Ji H., Liu G.Y., Mei H., Li K, & Chen T. (2018). miR-21-5p regulates type II alveolar epithelial cell apoptosis in hyperoxic acute lung injury. Molecular Medicine Reports, 17(4), 5796-5804.

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Selective Glucose Detection using Fe1@Au NPs

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The selectivity of Fe₁@Au NPs system for glucose was detected in solution containing glucose (0.1 mM) or interfering substances (dopamine (DA), L-cysteine, sucrose, fructose, ascorbic acid (AA), uric acid (UA), maltose, lactose and galactose, 1 mM). Specifically, the Fe₁@Au NPs (20 µL, 200 µg/ml) and glucose (20 µL, 1 mM) or other interfering chemicals (20 µL, 10 mM) were added into 96-well plates containing PBS buffer (140 μL, 10 mM, pH 7.2). Then, the mixed solution was incubated for 30 min at 37 °C. Next, the TMB (20 µL, 6 mM) was introduced to the above solution. Finally, the mixture was incubated for 10 min and absorbance was detected at 652 nm using a THERMO Varioskan Flash spectrophotometer.

Wang Q., Chen K., Jiang H., Chen C., Xiong C., Chen M., Xu J., Gao X., Xu S., Zhou H, & Wu Y. (2023). Cell-inspired design of cascade catalysis system by 3D spatially separated active sites. Nature Communications, 14, 5338.

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Kinetic Analysis of POD-mimicking Catalyst

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Kinetic experiments were monitored in a reaction volume of 200 μL HAc-NaAc bufer solution (pH = 4.0) containing 20 μg mL⁻¹ the POD-mimicking catalyst, TMB (0.1–1.2 mM) as a substrate and H₂O₂ (6 mM), or H₂O₂ (0.1–10 mM) as a substrate and TMB (0.6 mM). The mixture solution was incubated at room temperature for 10 min and then used for UV-vis absorbance measurement at 652 nm using a THERMO Varioskan Flash spectrophotometer. The Michaelis-Menten constant was calculated according to the Michaelis-Menten equation:

V_{0} = \frac{V_{\max} [S]}{(K_{m} + [S])}

The V₀ is the initial velocity, V_max corresponds to the maximum reaction velocity, which is monitored when the catalytic sites on the POD mimics are saturated with substrate. [S] is the initial substrate concentration, and Km is the Michaelis constant. The initial velocity V₀ was determined by according the slope of the kinetic curve in the initial phase, and the substrate concentration [S] was obtained at t = 0 s. The kinetic parameters Km and V_max were fitted according to Michaelis–Menten equation based on the calculated V₀ and [S].

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