Mouse brains were harvested in ice-cold PBS and homogenized with a tissue homogenizer (Kinematica, Luzern, Switzerland) at 15,000 rpm for 1 minute on ice. After centrifugation at 1500g for 2 minutes, the pellet was resuspended and incubated with solvent (DMSO), 19,20-EDP (100 μmol/L) or 19,20-DHDP (10 or 100 μmol/L) for 1 hour at room temperature with gentle shaking. Thereafter, samples were pelleted at 30,000g for 30 minutes, washed with PBS and centrifuged again (30,000g for 30 minutes) before being transferred into 4 mm MAS rotors and immediately measured. Spectra were recorded at 10 kHz MAS at a 1H Larmor frequency of 850 MHz with 64 scans and water presaturation. Sample temperatures were calibrated via KBr T1 relaxation time measurements. Samples were equilibrated for 10 minutes once the sensor reached the target temperature.
Tissue homogenizer
Manufactured by Kinematica
Sourced in Switzerland
The Tissue Homogenizer is a laboratory instrument designed to efficiently disrupt and homogenize a wide range of biological samples, including tissues, cells, and other solid materials. It utilizes high-speed rotation and specialized blades to thoroughly break down samples, ensuring efficient extraction and processing of their contents.
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3 protocols using tissue homogenizer
1
NMR Analysis of Mouse Brain Lipids
2
Rat Biomarker Quantification Protocol
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The ELISA kits for rat MTL, GAS, VIP, and ghrelin were purchased from Nanjing Jiancheng Technology Co., Ltd. (Nanjing, China). The other reagents were domestic and of analytical grade.
An ultrapure water meter (Millipore, USA), an Al104 electronic analytical balance (Shanghai METTLER TOLEDO Instrument Co., Ltd.), a Yp10002 Matou electronic balance (Shanghai Guangzheng Medical Instrument Co., Ltd.), a Tgl-5-a high-speed centrifuge (Shanghai Anting Analytical Instrument Co., Ltd.), a Varioskan flash full wavelength multifunctional microplate reader (Thermo, USA), a tissue homogenizer (Kinematica, Switzerland), and a 6890n gas chromatograph (Agilent, USA) were used.
An ultrapure water meter (Millipore, USA), an Al104 electronic analytical balance (Shanghai METTLER TOLEDO Instrument Co., Ltd.), a Yp10002 Matou electronic balance (Shanghai Guangzheng Medical Instrument Co., Ltd.), a Tgl-5-a high-speed centrifuge (Shanghai Anting Analytical Instrument Co., Ltd.), a Varioskan flash full wavelength multifunctional microplate reader (Thermo, USA), a tissue homogenizer (Kinematica, Switzerland), and a 6890n gas chromatograph (Agilent, USA) were used.
3
Pneumococcal Infection Model in Mice
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Streptococcus pneumoniae serotype 19F (strain BHN100) [18 (link)] was provided by Birgitta Henriques-Normark (Karolinska Institutet, Stockholm). Bacteria were grown in Todd-Hewitt yeast (THY) medium as previously described [19 (link)]. LPS- and IFN-γ-treated C57BL/6JRj mice and control groups were infected i.n. with 108S. pneumoniae 19F in 10 µl PBS 48 h after the first treatment. Mice were sacrificed 18 h post-infection and lung, trachea, NALT, and nasopharynx were homogenized using a tissue homogenizer (KINEMATICA AG, Switzerland). Samples were plated onto Columbia blood agar plates (BD Diagnostic Systems, Germany) and incubated over night at 37 °C, 5% CO2. CFU were counted to determine the bacterial burden.
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