The chemistry reagents (chemical grade) used in experiments was obtained from Sino Pharm Chemical Reagent Co. Ltd. (Shanghai, China). The real samples river water and tap water were obtained from the Shanghai University of Traditional Chinese Medicine, and sugar was purchased from a local supermarket (Shanghai, China). The chrysanthemum was purchased from Shanghai Kangqiao Chinese Medicine, grown in HangZhou, China. The analytical thin-layer chromatography (TLC) with pre-coated silica gel GF254 plates (Sino Pharm Chemical Reagent Co. Ltd., Shanghai, China) was used to detect any spots at 254 nm under UV light. 1H-NMR and 13C-NMR spectra were measured at 25 °C and referenced to tetramethyl silane (TMS) by a BRUKER AVANCE III spectrometer (Bruker, German). Electron ionization mass spectra (ESI-MS) was recorded on an Agilent 6460 triple quad LC-MS mass spectrometer (Agilent, Santa Clara, CA, USA). UV-Vis spectra and fluorescence spectra were recorded by Agilent 8454 UV-Vis spectrometer (Agilent) and Agilent G9800A fluorescence spectrophotometer (Agilent) respectively. Leica TCS-SP8 multiphoton illustrated the fluorescence images with the confocal microscope having a 63 × oil-immersion objective lens (Leica, Wetzlar, German).
Oil immersion objective lens
Manufactured by Leica
Sourced in Switzerland, Germany
The 63× oil-immersion objective lens is a high-magnification lens designed for use with Leica microscopes. It provides a magnification of 63× and is intended for use with oil immersion, which enhances image resolution and clarity. The lens is a core component for detailed microscopic examination and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Avance 3 spectrometer, by Bruker (2 mentions)
Agilent 6460 triple quad lc ms mass spectrometer, by Agilent Technologies (2 mentions)
Agilent 8454 uv vis spectrometer, by Agilent Technologies (2 mentions)
Agilent g9800a fluorescence spectrophotometer, by Agilent Technologies (2 mentions)
Tcs sp8 multiphoton, by Leica (2 mentions)
Precoated silica gel gf 254 plates, by Sinopharm (1 mentions)
Ms agilent 1100 series lc msd trap mass spectrometer, by Agilent Technologies (1 mentions)
Tensor 27 spectrometer, by Bruker (1 mentions)
Tcs sp8 confocal microscope, by Leica (1 mentions)
Fluorescence microscope, by Leica (1 mentions)
Las x software, by Leica (1 mentions)
Sp8 laser scanning confocal microscope, by Leica (1 mentions)
5 protocols using oil immersion objective lens
1
Characterization of Chrysanthemum Extracts
2
Synthesis and Characterization of Novel Compounds
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All chemicals (reagent grade) used were purchased from Sino Pharm Chemical Reagent Co., Ltd. (Shanghai, China). Reaction progress was monitored using analytical thin layer chromatography (TLC) on pre-coated silica gel GF254 (Qingdao Haiyang Chemical Plant, Qingdao, China) plates, and spots were detected under UV light (254 nm). Melting point was measured on an XT-4 micromelting point instrument and uncorrected. IR (KBr-disc) spectra were recorded by a Bruker Tensor 27 spectrometer (Bruker, German). 1H-NMR and 13C-NMR spectra were measured on a BRUKER AVANCE III spectrometer at 25 °C and referenced to tetramethyl silane (TMS) (Bruker, German). Mass spectra were obtained on an MS Agilent 1100 Series LC/MSD Trap mass spectrometer (Agilent, Santa Clara, CA, USA). UV-vis spectra were recorded on an Agilent 8454 UV-vis spectrometer (Agilent). Fluorescence spectra measurements were recorded on an Agilent G9800A fluorescence spectrophotometer (Agilent). Fluorescence images were obtained on a Leica TCS-SP8 multiphoton; a confocal microscope and a 63× oil-immersion objective lens was used (Leica, Switzerland). High-resolution EI mass spectra were recorded on an Agilent 6460 triple quad LC-MS mass spectrometer (Agilent).
3
Live-cell Mitochondrial Membrane Potential Imaging
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Live-cell microscopy was performed by transferring cells into selective medium with 3% glycerol at pH 7.4. For estimation of mitochondrial membrane potential, cells were stained with 0.5 µM TMRE for 15 min at 30°C and stained with 10 µg/ml 4′,6-diamidino-2-phenylindole (DAPI) for 5 min at 30°C. Finally, cells were immobilized on a thin agarose pad (1% [wt/vol] agarose in selective medium with 3% glycerol at pH 7.4). Microscopic pictures were performed by an EVOSFl Cell Imaging System (AMG, Germany) with a 100× oil immersion objective lens (NA 1.28; AMG, Germany) or a TCS SP8 confocal microscope with a 63× oil immersion objective lens (NA 1.4; Leica, Germany) and processed using Fiji (Schindelin et al., 2012 (link)).
4
Fluorescence Probe-based Sulfite Detection
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HepG2 cells came from the Chinese Academy of Sciences. The cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) added with 1% Penicillin-Streptomycin Solution (100×), 10% fetal bovine serum (FBS) at 37 °C under the atmosphere containing 5% CO2. Then transferred cells to confocal dishes and waited for 24 h to keep adaption.
To inspect the sensitivity of the probe, cells were incubated in 10 μM probe for 2 h at 37 °C with culture medium (90% DMEM, 10% FBS). To detect exogenetic sulfite, cells were pretreated with different concentrations of SO32− (0–200 μM) at 37 °C for 2 h and then incubated with the probe (10 μM) for 2 h. Before the detection achieved by the apparatus of Fluorescence Microscope and a 63× oil-immersion objective lens (Leica), cells needed to be washed by PBS for three times. The cells were stimulated by the intensity of wavelengths at 405 nm, 488 nm, and collected emission on 450–490 nm, 550–590 nm.
To inspect the sensitivity of the probe, cells were incubated in 10 μM probe for 2 h at 37 °C with culture medium (90% DMEM, 10% FBS). To detect exogenetic sulfite, cells were pretreated with different concentrations of SO32− (0–200 μM) at 37 °C for 2 h and then incubated with the probe (10 μM) for 2 h. Before the detection achieved by the apparatus of Fluorescence Microscope and a 63× oil-immersion objective lens (Leica), cells needed to be washed by PBS for three times. The cells were stimulated by the intensity of wavelengths at 405 nm, 488 nm, and collected emission on 450–490 nm, 550–590 nm.
5
Rapamycin-Induced 3D Imaging of HeLa Cells
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HeLa cells were grown in glass-bottom chamber and transfected with plasmids the day before fixation. Cells were stimulated with rapamycin for 30 min and fixed by 4% PFA before imaging. Three-dimensional imaging was performed with a laser-scanning confocal microscope sp8 (Leica) equipped with white laser and HyD detector array, using ×63 oil-immersion objective lens (Leica). The three-dimensional image stack was deconvoluted using the Lightning mode of LAS-X software (Leica).
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