Accuskan

Manufactured by Thermo Fisher Scientific

The AccuSkan is a laboratory instrument designed for the automated scanning and analysis of biological samples. It provides accurate and consistent measurements of various parameters, such as cell counts, viability, and morphology. The AccuSkan is a reliable tool for researchers and scientists working in the life sciences and biomedical fields.

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Lab products found in correlation

Pierce coomassie plus assay reagent, by Thermo Fisher Scientific (2 mentions) Multiskan fc, by Thermo Fisher Scientific (2 mentions) Cellstar 96 well microtiter plates, by Greiner (1 mentions) Agilent 7820a gc, by Agilent Technologies (1 mentions) Human il 6 quantikine elisa kit, by R&D Systems (1 mentions)

Close spelling variants of this product

AccuSkan™ GO UV/Vis Microplate Spectrophotometer AccuSkan FC microplate AccuSkan™ FC Filter‐Based Microplate Photometer AccuSkan FC microplate photometer AccuSkan FC plate reader Fisherbrand™ accuSkan™ GO UV/Vis Microplate Spectrophotometer AccuSkan FC AccuSkan™ GO UV/Vis AccuSkan Go spectrophotometer

5 protocols using accuskan

Outer Membrane Vesicle Protein Quantification

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A Bradford assay was used to determine outer membrane vesicle protein concentrations, as previously described (20 (link)). To generate a standard curve, bovine serum albumin (BSA) was diluted at 0 to 20 mg/mL in Pierce Coomassie Plus assay reagent (Thermo Fisher) to a final volume of 1 mL. Outer membrane vesicles were diluted at 2, 5, 10, 15, and 20 μL in reagent to a final volume of 1 mL. A microplate spectrophotometer (Fisherbrand AccuSkan) was used to measure the absorbance (OD₅₉₅) of the standard and samples in a 96-well plate (BrandTech). Protein concentrations were determined by comparing the optical densities of the samples to the standard curve plotted in Microsoft Excel, and final quantifications were graphed in GraphPad Prism 9. Experiments were reproduced three times from each outer membrane vesicle isolation, and one representative data set is reported.

Islam N., Kazi M.I., Kang K.N., Biboy J., Gray J., Ahmed F., Schargel R.D., Boutte C.C., Dörr T., Vollmer W, & Boll J.M. (2022). Peptidoglycan Recycling Promotes Outer Membrane Integrity and Carbapenem Tolerance in Acinetobacter baumannii. mBio, 13(3), e01001-22.

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Quantifying Outer Membrane Vesicle Proteins

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Bradford assay was used to determine outer membrane vesicle protein concentration, as previously described 44 (link) . To generate a standard curve, bovine serum albumin (BSA) was diluted 0 to 20 mg/mL in Pierce Coomassie Plus assay reagent (ThermoFisher) to a final volume of 1 mL. Outer membrane vesicles were diluted 2, 5, 10, 15, 20 L in reagent to a final volume of 1 mL. A microplate spectrophotometer (Fisherbrand AccuSkan) was used to measure the absorbance (OD595) of standard and samples in a 96-well plate (BrandTech). Protein concentrations were determined by comparing the optical densities of samples to the standard curve plotted in Microsoft Excel and final quantifications were graphed in GraphPad Prism 9. Experiments were reproduced three times from each outer membrane vesicle isolation, and one representative data set was reported.

Islam N., Kazi M.I., Kang K.N., Biboy J., Gray J., Ahmed F., Schargel R.D., Boutte C.C., Dörr T., Vollmer W., & Boll J.M. (2021). Peptidoglycan recycling contributes to outer membrane integrity and carbapenem tolerance inAcinetobacter baumannii.

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Antibiotic Susceptibility of Bacterial Pathogens

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Staphylococcus aureus USA 300 JE2, Staphylococcus aureus USA300 LAC, Escherichia coli K12 were provided by Dr. Bayles’ research lab and Pseudomonas aeruginosa PA01 was provided by Dr. Gus Wang’s lab (both at the Departament of Pathologhy and Microbiology of the University of Nebraska Medical Center – UNMC). The MICs of the PAs were studied using the broth microdilution method. Bacterial cultures were made by the direct colony suspension method to 1.5 × 10⁸ colony forming unit CFU/ml and dilute for 2 mL into 40 mL of Muller Hinton Broth (MHB) to a final concentration of ~10⁵ CFU/mL. A stock solution of each PAs was prepared in ultrapure water at 1 mg/ml concentration and pH was adjusted to 7. Then, serial dilutions were made in MHB, in Cellstar 96-well microtiter plates (Greiner, Bio-One). Each well was inoculated with 10 μL of bacterial cultures. The plates were incubated statically for 16–24 h at 37°C. The lowest concentration of PA that prevented bacterial growth was considered the MIC. The O.D value was set at 600 nm and was recorded with an AccuSkan, MultiSkan FC (Thermo Fisher Scientific). Vancomycin and Gentamicin were used as positive controls and media was used as negative control. The assay was performed in triplicate.

Rodrigues de Almeida N., Han Y., Perez J., Kirkpatrick S., Wang Y, & Sheridan M.C. (2019). Design, Synthesis, and Nanostructure-Dependent Antibacterial Activity of Cationic Peptide Amphiphiles. ACS applied materials & interfaces, 11(3), 2790-2801.

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Rumen Fluid Volatile Fatty Acid Analysis

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The pH was recorded using a digital pH meter after opening the bottles. Concentrations of volatile fatty acids (VFA) in ruminal fluid samples were determined in a liquid–liquid solvent extraction using ethyl acetate^{52 (link)} and analyzed by gas chromatography (Agilent 7820A GC, Agilent Technologies, Palo Alto, CA) using a flame ionization detector and a capillary column (CP-WAX 58 FFAP 25 m 0.53 mm, Varian CP7767, Varian Analytical Instruments, Walnut Creek, CA). Column temperature was maintained at 110 °C, and injector and detector temperatures were 200 and 220 °C, respectively.
Concentrations of NH₃-N in the rumen fluid were measured following the phenol-hypochlorite technique as described by Broderick and Kang^{53 (link)}. Absorbance was read in 96-well, flat bottom plates at 620 nm using a plate reader (AccuSkan, Thermo Fisher Scientific, Waltham, MA).

O. S. van Cleef F., B. Dubeux JC J.r., Wheeler C.S., V. García C.C., Ruiz-Moreno M., Sollenberger L.E., B. Vendramini J.M., DiLorenzo N, & Naumann H.D. (2022). Stable isotopes provide evidence that condensed tannins from sericea lespedeza are degraded by ruminal microbes. Scientific Reports, 12, 14318.

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Quantifying IL-1β and IL-6 Cytokines

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IL-1β and IL-6 measurements were made in the cell supernatants, by using Quantikine Human IL-1β/IL-1F2 ELISA Kit and Human IL-6 Quantikine ELISA Kit (R&D Systems) following manufacturer’s instructions. Samples were quantified using corrected values of 450 and 570 nm, reading absorbance in a microplate reader (Thermo Fisher Scientific, accuSkan). A 4-parameter logistic curve was used to describe the data. For each experiment at least 2 replicates were used for analysis.

Pathak S, & Vambutas A. (2022). NaCl exposure results in increased expression and processing of IL-1β in Meniere’s disease patients. Scientific Reports, 12, 4957.

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