For the Cell Counting Kit (CCK)-8 assay, keloid-derived fibroblasts (3,000 per well) were plated in 96-well plates. Following incubation for different durations (1, 2 and 3 days), cell viability was determined by adding 10 µl CCK-8 reagent to each well, according to the manufacturer's instructions (Abmole Bioscience, Inc.). The absorbance (450 nm) was determined using a microplate reader (Shenzhen Mindray Bio-Medical Electronics Co., Ltd.). For the colony formation assay, keloid-derived fibroblasts (3,000 per well) were plated in 6-well plates. After incubation at 37°C for two weeks, keloid-derived fibroblasts were fixed in 4% paraformaldehyde for 20 min at 25°C and then stained with 0.1% crystal violet (Beyotime Institute of Biotechnology) for 20 min at 25°C. Visible colonies were observed using a WMS-1033 Digital light biomicroscope at 200× magnification (Shanghai Wumox Optical Instrument Co., Ltd.). Images were captured using an EOS M50 digital camera (Canon, Inc.) and quantified by three observers.
Eos m50 digital camera
Manufactured by Canon
The EOS M50 is a compact, mirrorless digital camera produced by Canon. It features a 24.1-megapixel APS-C CMOS sensor and a DIGIC 8 image processor. The camera can capture high-quality still images and Full HD 1080p video. It supports a range of Canon EF-M and EF lenses.
Automatically generated - may contain errors
3 protocols using eos m50 digital camera
1
Cytotoxicity and Colony Formation in Keloid Fibroblasts
2
Ultrastructural Analysis of Plant Seedlings
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High-pressure freezing, freeze substitution, resin embedding, and ultramicrotomy were performed as described in Kang (Kang, 2010) . Seedlings were examined with a Canon EOS M50 Digital Camera equipped with fluorescence illumination to remove abnormal cotyledons before freezing. Frozen samples were freeze-substituted in anhydrous acetone with 1% OsO4 at -80°C for 24 hr. Excess OsO4 was removed at -80 o C by rinsing with precooled acetone. After being slowly warmed up to room temperature over 60 h, samples were separated from planchettes and embedded in Embed-812 resin (Electron Microscopy Sciences, USA; Cat. No. 14120). 80 nm thick sections of each time point were prepared with ultramicrotomy and then were examined with a Hitachi H-7650 TEM (Hitachi-High Technologies, Japan) operated at 80 kV.
3
High-Pressure Freezing for Ultrastructural Analysis
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High-pressure freezing, freeze substitution, resin embedding, and ultramicrotomy were performed as described in Kang (Kang, 2010) . Seedlings were examined with a Canon EOS M50 Digital Camera equipped with fluorescence illumination to remove abnormal cotyledons before freezing. Frozen samples were freeze-substituted in anhydrous acetone with 1% OsO4 at -80°C for 24 hr. Excess OsO4 was removed at -80 o C by rinsing with precooled acetone. After being slowly warmed up to room temperature over 60 h, samples were separated from planchettes and embedded in Embed-812 resin (Electron Microscopy Sciences; catalog no. 14120). 80 nm thick sections of each time point were prepared with ultramicrotomy and then were examined with a Hitachi 7400 TEM (Hitachi-High Technologies) operated at 80 kV.
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