All solid samples (n = 36, contains 3 replicates) were freeze-dried (Alpha 1-2 LD freeze-dryer; Martin Christ, Osterode am Harz, Germany) and passed through a sterile 2-mm sieve. The sieved samples were mixed with ultrapure water (1:5, wt/vol) to get a suspension. The supernatant pH of the suspension was determined using a multiparameter water quality detector (Hach, Loveland, CO, USA) (25 (link)). Dissolved anions and cations were measured with anionic chromatography (ICS-600; Thermo Scientific, USA) and inductively coupled plasma-optical emission spectrometry (ICP-OES) (iCAP 7600+; Thermo, USA), respectively, after filtration with 0.22-μm filters (47 (link)). The concentrations of CH4 and CO2 gases and the carbon isotope of CO2 ([δ13C]CO2) of cave air samples were measured by a high-precision carbon isotope analyzer (G2201-I; Picarro, USA) using cavity decay spectroscopy (cavity ring-down spectroscopy [CRDS]) (5 (link)) at the Institute of Karst Geology, Chinese Academy of Geological Sciences.
Alpha 1 2 ld freeze dryer
Manufactured by Martin Christ
Sourced in Germany, United Kingdom
The Alpha 1-2 LD is a freeze-dryer designed for laboratory use. It is capable of freeze-drying samples, with a maximum drying area of 0.18 m².
Automatically generated - may contain errors
Lab products found in correlation
Icap 7600, by Thermo Fisher Scientific (3 mentions)
Ics 600, by Thermo Fisher Scientific (3 mentions)
Nicolet 6700 ft ir, by Thermo Fisher Scientific (3 mentions)
Multiparameter water quality detector, by HACH (2 mentions)
Ultraturrax t 25 digital, by IKA Group (1 mentions)
Emitech sc7620, by Quorum Technologies (1 mentions)
Vega3, by TESCAN (1 mentions)
Sarcosine, by Merck Group (1 mentions)
Aas 18 5ml, by Merck Group (1 mentions)
L tryptophan, by Merck Group (1 mentions)
Chromeleon 7.2 sr4, by Thermo Fisher Scientific (1 mentions)
L asparagine, by Merck Group (1 mentions)
L glutamine, by Merck Group (1 mentions)
12 protocols using alpha 1 2 ld freeze dryer
1
Characterization of Cave Geochemistry
2
Freeze-Drying and FT-IR Analysis of BNC
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Before testing, samples were freeze-dried in an ALPHA 1–2/LD freeze dryer (Martin Christ GmbH, Osterode am Harz, Germany). Chemical analysis of variations in the structure of BNC produced under different conditions was performed by FT-IR with attenuated total reflectance mode (ATR). The spectra were recorded at a resolution of 8 cm−1, in the range of 4000 to 650 cm−1 using a Nicolet 6700 FT-IR (Thermo Fischer Scientific, Waltham, MA, USA). For each sample, 200 scans were taken.
3
Extraction and Antioxidant Evaluation of Chestnut Burs
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The chestnut spiny burs were collected in October in a chestnut grove (Montella, Avellino, Italy), air-dried and crushed by a blade mill (Grindomix RM 100, Retsch, Bergamo, Italy).
The chestnut burs hydroalcoholic extract was prepared as reported by Esposito et al. (2019) [6 (link)]. Briefly, a mixture of aqueous ethanol (50%, v/v) at 45 °C, was added to a sample (50 g) of dried chestnut burs. The mixture was homogenized by Ultra-Turrax (IKA ULTRA-TURRAX T25 digital, IKA-Werke GmbH & Co. KG, Staufen, Germany), at 10,000 rpm for 8 min, and left for 30 min under magnetic stirring. After filtration through a 45 μm mesh sieve and centrifugation (by Thermo Electron/ALC PK120-V1, Thermo Fisher Scientific, Waltham, MA, USA) for 5 min at 5000 rpm, the liquid extract was processed in a rotavapor (Heidolph Hei-VAP Value Digital Rotary Evaporator, Heidolph Instruments GmbH & Co. KG, Schwabach, Germany) to remove the ethanol. Subsequently, the extract was freeze-dried (Alpha 1–2 LD freeze dryer, Martin Christ, Osterode am Harz, Germany) to remove the residual aqueous portion until a dry extract (CSE) was obtained. The stable 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical was employed to determine the antiradical activity of CSE using the procedures previously described [23 (link),24 (link)].
The chestnut burs hydroalcoholic extract was prepared as reported by Esposito et al. (2019) [6 (link)]. Briefly, a mixture of aqueous ethanol (50%, v/v) at 45 °C, was added to a sample (50 g) of dried chestnut burs. The mixture was homogenized by Ultra-Turrax (IKA ULTRA-TURRAX T25 digital, IKA-Werke GmbH & Co. KG, Staufen, Germany), at 10,000 rpm for 8 min, and left for 30 min under magnetic stirring. After filtration through a 45 μm mesh sieve and centrifugation (by Thermo Electron/ALC PK120-V1, Thermo Fisher Scientific, Waltham, MA, USA) for 5 min at 5000 rpm, the liquid extract was processed in a rotavapor (Heidolph Hei-VAP Value Digital Rotary Evaporator, Heidolph Instruments GmbH & Co. KG, Schwabach, Germany) to remove the ethanol. Subsequently, the extract was freeze-dried (Alpha 1–2 LD freeze dryer, Martin Christ, Osterode am Harz, Germany) to remove the residual aqueous portion until a dry extract (CSE) was obtained. The stable 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical was employed to determine the antiradical activity of CSE using the procedures previously described [23 (link),24 (link)].
4
Characterization of Cave Geochemistry
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All solid samples (n = 36, contains 3 replicates) were freeze-dried (Alpha 1-2 LD freeze-dryer; Martin Christ, Osterode am Harz, Germany) and passed through a sterile 2-mm sieve. The sieved samples were mixed with ultrapure water (1:5, wt/vol) to get a suspension. The supernatant pH of the suspension was determined using a multiparameter water quality detector (Hach, Loveland, CO, USA) (25 (link)). Dissolved anions and cations were measured with anionic chromatography (ICS-600; Thermo Scientific, USA) and inductively coupled plasma-optical emission spectrometry (ICP-OES) (iCAP 7600+; Thermo, USA), respectively, after filtration with 0.22-μm filters (47 (link)). The concentrations of CH4 and CO2 gases and the carbon isotope of CO2 ([δ13C]CO2) of cave air samples were measured by a high-precision carbon isotope analyzer (G2201-I; Picarro, USA) using cavity decay spectroscopy (cavity ring-down spectroscopy [CRDS]) (5 (link)) at the Institute of Karst Geology, Chinese Academy of Geological Sciences.
5
Soil Physicochemical Analysis Protocol
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All samples were freeze-dried (Alpha 12 LD freeze-dryer; Martin Christ, Osterode am Harz, Germany) and passed through a sterile 200-mesh sieve. Total organic carbon (TOC) content and C/N ratio were analyzed with a C-S analyzer (EA 4000, Analytik Jena AG, Jena, Germany) after acidification with 3 M HCl. One gram soil samples were mixed with 5 mL ultrapure water followed by 10 min vortex and subsequent centrifugation at 6,800 × g for 10 min (Yun et al., 2016b (link)). Filtrates (0.22 μm membrane) were used for the pH measurement and dissolved ion analysis. The pH was measured with a multi-parameter water quality detector (HACH, Loveland, CO). The analysis of dissolved anions and cations was performed using anionic chromatography (ICS-600, Thermo Scientific, Waltham, MA) and ICP-OES (iCAP 7,600+, Thermo Scientific), respectively.
6
Scanning Electron Microscopy Analysis of Solidified Implants
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The inner/surface morphologies of completely solidified implants were examined using a VEGA3 TESCAN (TESCAN, Brno, Czech Republic) scanning electron microscope (SEM). After rinsing in water carefully and then freeze-drying (Alpha 1-2 LD Freeze dryer, Christ, UK), the samples were treated with liquid nitrogen and then crushed. Samples were relocated onto metal stubs by double-adhesive conductive tape and sputter-coated with platinum using an EMITECH SC7620 sputter coater (Quorum Technologies Ltd., Laughton, United Kingdom). The voltage was set to 3.0 kV for SEM observation.
7
Nanostructured Lipid Carriers for Herbal Extracts
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The nanostructured lipid carriers encapsulating DSG and/or wild yam extract (Yam) were obtained using high shear homogenization and high-pressure homogenization procedures applied successively to an o/w emulsion as described by Lacatușu et al. [19 (link)]. The lipid phase consisting in a blend of solid lipids (GMS, CP) and vegetable oils (EPO or SOY) and the aqueous phase (which contains 2.5% surfactant) were mixed under continuous stirring and were kept for 20 min at 73 °C. Proportions established between lipid matrix and herbal bioactive (DSG and/or wild yam extract) were chosen following the optimization step (Table 1 ). Afterwards, the pre-emulsion was submitted to the HSH (High Shear Homogenizer PRO250, Oxford, CT, USA) at 12,000 rpm for 1 min and HPH (APV 2000 Lab Homogenizer, Lubeck, Germany) for 6 homogenization cycles at 500 bars. The obtained nanodispersions were cooled at room temperature, stored overnight at −25 °C, and freeze dried by lyophilization (−55 °C, 54 h, using a Martin Christ Alpha 1–2 LD Freeze Dryer, Osterode am Harz, Germany).
8
Morin-Infused Hydrogel Scaffolds
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Hydrogel scaffolds were formulated incorporating varying concentrations of morin. 400 mg each of KER and PSH, and varying weights of morin (0, 100, and 200 mg) were dissolved in 18 ml of distilled water. 1 ml each of 15% STMP and 30% NaOH were added to the admixture which was then poured into Teflon molds, kept at −20 °C for 12 h, and lyophilized (Christ Alpha 1-2 LD Freeze Dryer, UK). The freeze-dried product was washed thoroughly with distilled water to remove the NaOH, and subsequently re-lyophilized to obtain porous hydrogel scaffolds. The scaffolds obtained have been termed as PSH + KER, PSH + KER + 0.5% MOR, and PSH + KER + 1% MOR, based on the morin concentration of the preparatory solution.
9
Frog Secretion Extraction and Preservation
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The frogs Amolops hainanensis were all mature adults and captured in the field in China. The secretion acquisition procedure was performed as described previously [31 (link)]. After being frozen in liquid nitrogen and lyophilized in an Alpha 1-2/LD freeze dryer (Martinchrist, Osterode, German), the sample was stored at −20 °C prior to use. This study was approved by the Nanjing University of Chinese Medicine Ethical Review Board—Approval Code: SYXK (S.U.) 2018-0048, 26 October 2018.
10
Quantitative Amino Acid Profiling
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Amino acid profiles were quantified using a modified version of the procedure described by Fürst, Pollack, Graser, Godel, and Stehle (1990) (link). Leaf material was lyophilized using an Alpha 1-2 LD+ freeze dryer (Christ, Osterode, Germany). About 10 mg plant powder was arly from 0 to 4.4 min to 71.5:28.5, followed by an isocratic step for 0.3 min. From 4.7 to 6.9 min, solvent A reduced further to 43%, followed again by an isocratic step till 9 min. At 9.8 min, the composition reached 0:100 (A:B) and remained for 5.7 min, to wash the column. At 15.7 min, the initial composition of 98:2 (A:B) was used for 4.3 min to re-equilibrate the column for the next run. Amino acids were detected using a Fluorescence detector 3400 RS and UV-Detector 3100 (Thermo Fisher, Dreieich, Germany). Primary amino acids were excited with 337 nm wavelength, whereas the emitted light at 442 nm was measured. Secondary amino acids were excited with 266 nm, and the emitted signals detected at 305 nm. Standards were prepared using the amino acid standard solution from Sigma-Aldrich (AAS18-5ML), stacked with L-Asparagine, L-Glutamine, gamma-Aminobutyrate, beta-Aminobutyrate, L-Tryptophan and Sarcosine (each from Sigma-Aldrich, Hamburg, Germany). Evaluation of the peak areas was done with the software Chromeleon 7.2 SR4 (Thermo Fisher, Dreieich, Germany).
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