The human CRC cell lines SW620, DLD1, and HCT116, and colorectal mucosa cell line FHC were purchased from the cell bank of the Chinese Academy of Sciences (Shanghai, China) and cultured in RPMI 1640 medium (Invitrogen Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Gibco), 100 U/ml penicillin, and 50 mg/ml streptomycin. All cells were incubated at 37 °C with a humidified atmosphere of 5% CO2. The morphology of the CRC cells was observed using phasemicroscopy (Leica,Wetzlar, Germany).
Phase microscopy
Manufactured by Leica
Sourced in Germany
Phase microscopy is an optical microscopy technique that allows for the visualization of transparent or low-contrast specimens by enhancing the contrast between different structures within the sample. The core function of phase microscopy is to convert phase shifts in the light passing through the specimen into differences in brightness or contrast, making it possible to observe and study these structures in detail.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Vimentin, by Abcam (1 mentions)
Tetramethylrhodamine isothiocyanate conjugated secondary antibodies, by Thermo Fisher Scientific (1 mentions)
E cadherin, by Abcam (1 mentions)
N cadherin, by Abcam (1 mentions)
β catenin, by Abcam (1 mentions)
Hepg2, by American Type Culture Collection (1 mentions)
4 protocols using phase microscopy
1
Culturing Human Colorectal Cancer Cell Lines
2
Cultivation of Human Liver Cancer Cells
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The human HCC cell line HepG2 cells (obtained from the Cell Bank of the Chinese Academy of Sciences, Shanghai, China) and HCCLM3 cells [47 (link)] (human HCC cell lines with high metastatic potential, established at Liver Cancer Institute, Fudan University, Shanghai, China) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen, Carlsbad, CA) containing 10 % fetal bovine serum (FBS) and 100 U ml−1 penicillin and 50 mg ml−1 streptomycin. All cells were incubated at 37 °C with a humidified atmosphere of 5 % CO2. The morphology of the HCC cells was observed using phase microscopy (Leica).
3
Immunofluorescence Analysis of EMT Markers
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The morphologies of the HCC cells were observed using phase microscopy (Leica). The immunofluorescence staining was carried out as previously described with some changes [29] (link). Cultured cells were grown in cell culture dishes and then washed and fixed. Cells were then incubated with primary antibodies to E-cadherin, N-cadherin, vimentin and β-catenin (1∶200, Abcam, Hong Kong), and anti-mouse FITC, anti–rabbit FITC and/or tetramethyl rhodamine isothiocyanate–conjugated secondary antibodies (Invitrogen). The fluorescent images were taken using a confocal microscope (Olympus).
4
Hepatocellular Carcinoma Cell Lines
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Six HCC cell lines, HepG2 (American Type Culture Collection), Huh7 (Japanese Cancer Research Bank), SMMC7721 (the Second Military Medicine College, Shanghai, China), MHCC97L, MHCC97H, and HCCLM3 (the Liver Cancer Institute of Zhongshan Hospital, Shanghai, China) were used in this study. One normal liver cell line, L0-2 (American Type Culture Collection) served as a control. All cell lines were incubated in high glucose Dulbecco’s modified Eagle medium (DMEM; Invitrogen), supplemented with 10% fetal bovine serum (FBS; Invitrogen) at 37 °C under 5% CO2 in humidified incubator. The morphology of HCC cells was assessed by phase microscopy (Leica).
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