To determine the BacFL31 impact on cells membrane permeability, the released extracellular ions K+ were measured. L. monocytogenes ATCC 19117 cells were centrifuged and resuspended in sterile water to reach 107 CFU/mL of cell concentration. BacFL31 was added at a final concentration of 1 X MIC to the cells suspensions. After various time intervals (15, 30, 45, 60, and 75 min), the control (without BacFL31 addition) and the treated samples (2 mL for each essay) were centrifuged at 7000 g for 10 min. The supernatants were subjected to measurement of the released potassium ions by an Analyst 200 atomic absorption spectrometer (Perkin Elmer).
Analyst 200 atomic absorption spectrometer
Manufactured by PerkinElmer
The Analyst 200 Atomic Absorption Spectrometer is a laboratory instrument designed to measure the concentration of specific elements in a sample. It uses the principles of atomic absorption spectroscopy to analyze the elemental composition of materials.
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3 protocols using analyst 200 atomic absorption spectrometer
1
Measuring BacFL31 Impact on Cell Membrane
2
Characterization of Reactive Orange-16 Dye
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All chemicals and materials were used without further purification. Solutions of the Reactive Orange-16 dye (RO-16) (Sigma-Aldrich) with an assay of 50% were prepared by dissolving the weighed solid in distilled water.
The IR spectra was collected on a Bruker Alpha ATR-FTIR spectrometer at a resolution of 4 cm−1. Samples were analysed after normalization of the spectra (allows for differences in sample loading and concentration) at pH = 3. Iron content was determined on PerkinElmer Analyst 200 Atomic Absorption Spectrometer after digestion of the sample with Conc. HCl at 140 °C. UV-VIS measurements were carried out on an Evolution 220 UV-Visible spectrophotometer by Thermo Fisher Scientific at bandwidth of 2 nm. The SEM images were obtained from a Carl Zeiss EVO 15 and the EDX on an Oxford instrument Xmax 80 mm2. The EPR measurements were done on an EMXnano from Bruker at a g factor of 4.00. The XRD measurements were done on a Bruker XRD D2 PHASER 2theta/scan Cu tube with 1.54184 Å with Lynexy (ID mode) detector run from 10 to 80 2theta and resolved on a Diffrac.Suite EVA software (VERSION 4.2.1.10).
The IR spectra was collected on a Bruker Alpha ATR-FTIR spectrometer at a resolution of 4 cm−1. Samples were analysed after normalization of the spectra (allows for differences in sample loading and concentration) at pH = 3. Iron content was determined on PerkinElmer Analyst 200 Atomic Absorption Spectrometer after digestion of the sample with Conc. HCl at 140 °C. UV-VIS measurements were carried out on an Evolution 220 UV-Visible spectrophotometer by Thermo Fisher Scientific at bandwidth of 2 nm. The SEM images were obtained from a Carl Zeiss EVO 15 and the EDX on an Oxford instrument Xmax 80 mm2. The EPR measurements were done on an EMXnano from Bruker at a g factor of 4.00. The XRD measurements were done on a Bruker XRD D2 PHASER 2theta/scan Cu tube with 1.54184 Å with Lynexy (ID mode) detector run from 10 to 80 2theta and resolved on a Diffrac.Suite EVA software (VERSION 4.2.1.10).
3
Elemental Analysis of Plant Tissues
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Plant samples were prepared by mixing equal quantities of root and shoot, oven drying at 65 °C for 72 h and powdering and sieving with 0.5 mm mesh. The Micro-Kjeldahl method was used for the estimation of total nitrogen (N) [35 ]. Phosphorus (P) content was determined using an ammonium molybdate assay wherein plant tissues were digested in HNO3 and HClO4 [36 ]. Potassium (K) was determined using a flame photometer [37 ]. Ca (%), Mg (%), were estimated by atomic absorption spectrophotometry (Analyst 200 Atomic Absorption Spectrometer, PerkinElmer) [38 (link)] and total sulphur (%) was determined by flame emission photometry and the turbidimetric method, respectively, in the di-acid extract [39 (link)].
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