To determine the osteogenic differentiation of BMSCs transfected with si-Wnt2b or ov-Wnt2b, the cells were seeded in 48-well plates and incubated at 37 °C with 5% CO2 and saturated humidity. The BMSCs were stained at days 3, 7, and 10 after transfection and osteogenic induction. The calcium deposit was stained with 1% alizarin red staining kit and pictures were taken by microscopy (TS2-S-SM, Nikon, Japan). The osteogenic differentiation of BMSCs transfected with piR-63049 mimics or inhibitor was detected by ALP stain using the BCIP/NBT Alkaline Phosphatase Color Development Kit (Beyotime Biotechnology, China) at day 7 after transfection and osteogenic induction, according to the protocol. Then the cells were imaged using a Canon EOS 350D digital camera, placed vertically in a stable manner over a white background. Alkaline Phosphatase Assay Kit (Beyotime Biotechnology, China) was used to detect the ALP activity and the manufacturer's instruction.
Eos 350d digital camera
Manufactured by Canon
Sourced in United Kingdom
The EOS 350D is a digital SLR camera produced by Canon. It features a 8.0-megapixel CMOS sensor, a 3-point autofocus system, and the ability to record images in JPEG and RAW file formats. The camera supports a range of interchangeable lenses and has a compact, lightweight body design.
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2 protocols using eos 350d digital camera
1
Osteogenic Differentiation of BMSCs via Wnt2b and piR-63049
2
Synthesis and Characterization of Functionalized Nanomaterials
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ρ-Phenylenediamine (ρ-PDA), petroleum ether (PE), silica gel (xSiO2·yH2O), ethyl acetate (EA), citric acid (CA), ethylenediamine (EDA), disodium hydrogen phosphate (Na2HPO4·12H2O), sodium dihydrogen phosphate (NaH2PO4·2H2O), sodium chloride (NaCl), potassium chloride (KCl), ethanol, quartz sand, deionized water, and securities paper were purchased from Aladdin Ltd. All chemicals were used as received and did not require further purification. Deionized water (18.2 MΩ cm) was produced by a Millipore water purification system.
A DZF-6020 vacuum dryer (Shanghai Xinmiao Co., Ltd.), AL-204-IC electronic analysis balance (METTLER-TOLEDO), HP Printer 2050 (China Hewlett–Packard Co., Ltd.), KQ-300DE ultrasound cleaner (Kunshan Instrument Co., Ltd.), H1650-W centrifuge (Hunan Instrument Co., Ltd.), FP-6500 fluorescence spectrometer (Jusco (Shanghai) Trading Co., Ltd), UV-3600 ultraviolet-visible spectrometer (Shimadzu Instruments and Equipment Co., Ltd.), TEM-2100 transmission electron microscope (Japan Electronics Co., Ltd.), FT-IR-5700 (Thermo Co., USA), X-ray photoelectron spectroscope (Thermo Fisher, UK), H-G lamp, Canon EOS 350D digital camera, and USB portable UV lamp (365 nm) were used.
A DZF-6020 vacuum dryer (Shanghai Xinmiao Co., Ltd.), AL-204-IC electronic analysis balance (METTLER-TOLEDO), HP Printer 2050 (China Hewlett–Packard Co., Ltd.), KQ-300DE ultrasound cleaner (Kunshan Instrument Co., Ltd.), H1650-W centrifuge (Hunan Instrument Co., Ltd.), FP-6500 fluorescence spectrometer (Jusco (Shanghai) Trading Co., Ltd), UV-3600 ultraviolet-visible spectrometer (Shimadzu Instruments and Equipment Co., Ltd.), TEM-2100 transmission electron microscope (Japan Electronics Co., Ltd.), FT-IR-5700 (Thermo Co., USA), X-ray photoelectron spectroscope (Thermo Fisher, UK), H-G lamp, Canon EOS 350D digital camera, and USB portable UV lamp (365 nm) were used.
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