The tissue concentrations of C and N (w/w; presented as C% or N%) of the germinants grown on the unlabelled germination medium, was determined using an elemental analyser (Flash EA 2000, Thermo Fisher Scientific, Bremen, Germany). The 15 N content of the germinants, grown on 15 N-labelled germination media, was determined using an isotope ratio mass spectrometer (IRMS; DeltaV, Thermo Fisher Scientific, Bremen, Germany) interfaced to the element analyser (Flash EA 2000) . Sample weight used for elemental analysis was 2 mg (± 10%) DW.
Flash ea 2000
Manufactured by Thermo Fisher Scientific
Sourced in Germany, United States
The Flash EA 2000 is an elemental analyzer designed for the rapid and accurate determination of carbon, hydrogen, nitrogen, and sulfur content in a wide range of organic and inorganic sample types. It utilizes combustion and gas chromatography techniques to provide reliable and reproducible results.
Automatically generated - may contain errors
Lab products found in correlation
Delta 5, by Thermo Fisher Scientific (6 mentions)
Delta 5 plus, by Thermo Fisher Scientific (3 mentions)
Delta 5 advantage irms, by Thermo Fisher Scientific (3 mentions)
Fesem vp scanning electron microscope, by Zeiss (2 mentions)
Cary 630 atr ftir spectrophotometer, by Agilent Technologies (2 mentions)
Microlab expert v 1, by Agilent Technologies (2 mentions)
Quantachrome autosorb iq porosity equipment, by Anton Paar (2 mentions)
Conflo 4, by Thermo Fisher Scientific (1 mentions)
Delta 5 advantage isotope ratio mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Isotope ratio mass spectrometer deltav, by Thermo Fisher Scientific (1 mentions)
Winrhizo, by Regent Instruments (1 mentions)
Delta 5 advantage, by Thermo Fisher Scientific (1 mentions)
13c glucose, by Merck Group (1 mentions)
Mat253plus, by Thermo Fisher Scientific (1 mentions)
Mat 253 irms, by Thermo Fisher Scientific (1 mentions)
Mas 200 r, by Thermo Fisher Scientific (1 mentions)
Finnigan delta plus, by Thermo Fisher Scientific (1 mentions)
Mas probe, by Bruker (1 mentions)
Av 300, by Bruker (1 mentions)
36 protocols using flash ea 2000
1
Germinant C and N Tissue Analysis
2
Stable Isotope Analysis of Ground Samples
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Ground samples were packed into 5 x 8 mm ultra-clean tin capsules and analysed using an elemental analyser (EA Flash 2000 from ThermoFisher Scientific) coupled with an isotope ratio mass spectrometer (Delta V Plus from ThermoFisher Scientific) at the stable isotope platform of the Pole Spectrométrie Océan at the University of Bretagne Occidentale (Brest, France). Stable isotope ratios were reported in the standard δ notation as units of parts peer thousands (‰) relative to the international reference standard: δX=[(RSample/RStandard)-1] * 10 3 where X is 13 C and 15 N and R is the corresponding ratio of 13 C/ 12 C and 15 N/ 14 N. Reference standard used were Vienna-Pee Dee Belemnite for 13 C and atmospheric N2 for 15 N (precision: 0.1).
3
PM10 Total Organic Carbon Measurement
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In this study, the total organic carbon (TC) is defined as the sum of elemental carbon (EC) and organic carbon (OC) [58 (link),59 (link)]. For EC measurements, the HCL fumigated filters were treated following the chemo-thermal oxidation method (CTO – 375) [60 (link),61 (link)]. The suitability of the CTO – 375 method and its comparison with other protocols are discussed by several workers [62 (link),63 (link)]. The filters were heated at 400 °C in active airflow for 24 h to remove the organic C fractions. The concentrations and isotopic composition (δ13C) of TC and EC were measured using an elemental analyzer; EA (Flash 2000; Thermo Fisher Scientific, Germany) connected to an Isotope Ratio Mass Spectrometer; IRMS (Delta V, Thermo Fisher Scientific, Germany). The IAEA cellulose standard (IAEA–CH– 3; δ13C = −24.7 ‰; C contents = 44 %) was used as laboratory standard. The analytical precision for repeat measurements of standards were better than 10 % (for C content) and 0.1 ‰ (for C isotopic composition). The sample preparation and stable isotopic measurements of TC and EC were conducted at Physical Research Laboratory, Ahmedabad. The fraction of PM10 was estimated using analytical protocols published elsewhere [34 (link),64 (link),65 (link)].
4
Microbiome Elemental Characterization
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The total amounts of C and N, and the corresponding isotope ratios of C (13C/12C) and N (15N/14N) in the microbial pellets, plants, and the endpoint pore water were analyzed with elemental analyzer (EA)-irMS (EA Flash 2000, Thermo; ConFlo IV, Thermo; Delta V Advantage IRMS, Thermo, Bremen, Germany). All calibration curves for C and N concentrations exhibited R2 > 0.99.
5
Decomposition and Nutrient Dynamics
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At 42 days after laying (DAL), we randomly sampled three Experimental bags of each residue/biomass under each tree. This was repeated during each sampling time at 83, 126, 216 and 277 DAL. During each sampling time, the decomposing residue/biomass were carefully removed from their Experimental bags and cleaned of all soil and plant debris; whenever necessary, the recovered Experimental bags were rinsed in water for 1-2 minutes to remove soil particles. The dry weight of the remaining residue/biomass was recorded after oven-drying at 60 o C for 72 hours to a constant mass. For the nutrient analyses, the dried residue/biomass was crushed through a 2mm sieve. Each sample was then milled into a homogenous powder, using a ball-and-capsule vibrating mill.
The carbon and N concentration of the samples was determined by encapsulating 2.0 -2.5 mg of the milled material in a tin cup and analyzed using the Continuous flow isotopic ration mass spectrometer, EA-Flash 2000 ThermoFisher Scientific instrument. The Ca, Mg, Mn and B concentrations were determined by ICP-OES (according to the EPA Method 6010C-ICP-OES (EPA 2007) after microwave digestion with nitric acid according to EPA Method 3052 (EPA 1996) .
The carbon and N concentration of the samples was determined by encapsulating 2.0 -2.5 mg of the milled material in a tin cup and analyzed using the Continuous flow isotopic ration mass spectrometer, EA-Flash 2000 ThermoFisher Scientific instrument. The Ca, Mg, Mn and B concentrations were determined by ICP-OES (according to the EPA Method 6010C-ICP-OES (EPA 2007) after microwave digestion with nitric acid according to EPA Method 3052 (EPA 1996) .
6
Collagen Extraction from Bone Fragments
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Collagen samples were extracted from the bones by gelatinization using methods improved from previous studies (83 (link), 84 (link)). The bone fragments were soaked in 0.4 M HCl for 48 h at 4 °C to remove hydroxyapatite, after which they were soaked in 0.1 M NaOH to remove humic material. Finally, the remains were heated in aqueous HCl (pH 4) at 90 °C for 48 h. They were then filtered and freeze-dried to produce collagen. The stable carbon and nitrogen isotope compositions of the collagen were determined using an EA–IRMS system (Flash 2000 EA coupled to a Delta V Advantage IRMS, Thermo Fisher Scientific) at the University Museum of the University of Tokyo. The atomic C/N ratio of collagen was expected to be in the range of 2.9 to 3.6 (85 ), and data for samples outside this range were excluded from the analysis.
7
Seafood Stable Isotope Analysis
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For the stable isotope analyses, approximately 500 mg of the freeze dried seafood samples was defatted with a mixture of MeOH/chloroform (2+1) and acetone by vigorous shaking. The solvent layer was removed after centrifugation. After evaporating the residual solvent at 55 o C, approximately 0.4 mg of defatted samples was enclosed in a tin capsule and analyzed by elemental analyzer-isotope ratio mass spectrometry (Thermo Fisher Scientific Flash 2000 EA coupled to a Delta V Advantage IRMS, Bremen, Germany). Stable isotope ratios were expressed as δ 13 C and δ 15 N according to the ordinal notation by the following equation.
where R represents 15 N/ 14 N or 13 C/ 12 C. Isotope ratios are expressed in per mil (‰) relative to the ratio of international reference standards, i.e., Pee Dee Belmnite for carbon and atmospheric N 2 for nitrogen. Analytical errors (1σ) for nitrogen and carbon were <0.2‰ based on the results of ten replicate analyses of reference glycine, alanine and histidine (SI Science Co., Ltd).
where R represents 15 N/ 14 N or 13 C/ 12 C. Isotope ratios are expressed in per mil (‰) relative to the ratio of international reference standards, i.e., Pee Dee Belmnite for carbon and atmospheric N 2 for nitrogen. Analytical errors (1σ) for nitrogen and carbon were <0.2‰ based on the results of ten replicate analyses of reference glycine, alanine and histidine (SI Science Co., Ltd).
8
Biomass and Nitrogen Content Quantification
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Plants were grown under sterile and long-day conditions for 19 days on N-free MS medium, 0.5 mM L-Gln, 1% (w/v) agar and 0.5% (w/v) sucrose, buffered to pH 5.8 using 7.7 mM MES. The seedlings were dried at 60 °C and subsequently weighed. A minimum of four replicates were used for biomass determination. Afterwards, seedlings were homogenized for determination of total N content. Partially, samples were pooled to obtain a minimum of three biological replicates. The analysis was performed using an Elemental Analyzer—Isotope Ratio Mass Spectrometer (EA-IRMS) (EA: Flash EA 2000, IRMS: Delta V, both from Thermo Fisher Scientific) (Werner et al. 1999 (link)). Data presented are mean values ± SE (P < 0.05, one-way ANOVA with an additional Dunnett’s test, n = 3). Black stars indicate the comparison between lht1, aap5 and lht1 aap5 with the control Col-0, as their genetical background is Col-0. T1:3 and T4:4 were compared with lht1 aap5, as their genetical background is lht1 aap5 (gray stars).
9
Quantification of Plant Biomass P, N, C
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For analysis of P concentration in plant biomass, samples of shoots or roots (0.1 g each) were incinerated at 550°C for 12 h, the ashes were extracted with 1 mL boiling concentrated HNO3, and the extracts were then made up to 50 mL with ultrapure (18.2-MΩ) water. Phosphorus concentration in the acid extracts was measured spectrophotometrically by the malachite green method (90 (link)). The N and C concentrations in the plant biomass (2-mg ground samples) and substrates (20-mg ground samples), as well as the isotopic composition of N in the same samples, were analyzed using the Flash EA 2000 elemental analyzer coupled to a Delta V Advantage isotope ratio mass spectrometer (Thermo Fisher Scientific, Bremen, Germany).
10
Nanomaterial Characterization by Advanced Techniques
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The specific surface area and pore distribution were investigated using a Quantachrome Autosorb-IQ porosity equipment (Quantachrome Instruments, Boynton Beach, FL, USA) by Brunauer–Emmett–Teller (BET) and Barrett–Joyner–Halenda (BJH) methods. Prior to analysis, the nanomaterials were degassed at 373–393 K. The morphology of all samples (before and after adsorption) was investigated using a FESEM VP Scanning Electron Microscope (Carl Zeiss, Oberkochen, Germany) at 0.8 nm resolution, 2.5 nm VP mode and 30 kV; also, EDS was applied for the determination of metal content. Functional groups were elucidated using a Cary 630 ATR-FTIR spectrophotometer (Agilent Technologies, Inc., Santa Clara, CA, USA). Prior to analysis, the samples were ground in an agate mortar and dried at 80 °C under vacuum. The FTIR spectra were registered between 4000 and 400 cm−1 (32 scans at a resolution of 8 cm−1 and a threshold of 0.002), and MicroLab Expert v.1.0.0 Software (Agilent Technologies, Inc., Santa Clara, CA, USA) was used for the interpretation.
The elemental analysis was achieved by combustion and pyrolysis coupled with gas chromatography (Flash EA2000, Thermo Scientific, Waltham, MA, USA) [23 (link)].
The elemental analysis was achieved by combustion and pyrolysis coupled with gas chromatography (Flash EA2000, Thermo Scientific, Waltham, MA, USA) [23 (link)].
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