PCa cells were inoculated into dishes containing 10 mL preheated culture medium, and evenly dispersed by gentle rotation, followed by culture, with the culture medium changed once every 2–3 days. The culture was halted once the clone in the dish was visible to the naked eyes. After removal of supernatant, the cells were washed, and fixed. The fixed cells were stained with an appropriate amount of GIMSA (Invitrogen, USA) for 10–30 min. The number of cell clones was counted under an inverted microscope (Leica DMi8-M, Co. Ltd., Solms, Germany), and the colony formation rate was calculated using the formula: the number of cell clones/the number of inoculated cells × 100%.
Gimsa
Manufactured by Thermo Fisher Scientific
Sourced in United States
The GIMSA is a laboratory equipment designed for sample analysis. It utilizes an advanced imaging system to capture high-resolution images of samples. The core function of the GIMSA is to provide visual data for researchers and scientists to analyze their samples effectively.
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Lab products found in correlation
4 protocols using gimsa
1
Colony Formation Assay for PCa Cells
2
Colony Formation Assay for MM Cells
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The MM cells in the logarithmic growth phase were detached with 0.25% trypsin and titrated gently to single cells. The number of living cells was counted, and the cell density was adjusted to 1 × 106 cells/mL. Each group of cells was inoculated with 500 cells per well in a 24-well plate containing 1 mL culture medium pre-heated at 37°C and then placed at 37°C in a 5% CO2 incubator for 2–3 weeks. The medium was renewed every 2–3 times. When the visible colonies appeared in the dish, the culture was terminated. The MM cells were fixed with 5 mL of 4% paraformaldehyde for 15 min. MM cells was added with GIMSA (Invitrogen) and stained for 10–30 min. Imaging and cell counting were performed under an inverted microscope (Leica DMi8-M, Germany). The clone formation rate (%) = number of clones formed/number of cells inoculated × 100%.
3
Cell Clone Quantification Protocol
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The quantities of living cells were counted. ESCC cells (1 × 106 cells/mL) were cultured in dishes containing 10 mL preheated medium at 37°C with 5% CO2 for 2–3 weeks. When visual clones appeared, cells were collected and fixed. After the fixative solution was removed, GIMSA (Invitrogen) was added into cells to stain the nuclei for 10–30 min. Staining solution was slowly washed off with running water. The plate was placed under an inverted microscope to count quantities of cell clones.
4
Clonogenic Assay for ESC410 Cells
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ESC410 cells at the logarithmic growth phase were digested with 0.25% trypsin and gently dispersed into single cells. The viable cells were counted and the cell density was adjusted to 1 × 106 cells/mL. Cells were seeded into a dish containing 10 mL of 37°C preheated culture solution at a gradient density of 50, 100, and 200 cells/dish, and gently rotated to disperse the cells. The cells were cultured in a 37°C incubator under 5% CO2 for 2-3 weeks. When there were visible clones in the petri dish, the culture was concluded. The supernatant was discarded and the cells were carefully rinsed twice with PBS. The cells were fixed with 5 mL of 4% paraformaldehyde, and then the fixative was removed. The cells were stained using an appropriate amount of GIMSA (Invitrogen) for 10-30 min and dried in the air after the staining solution had been slowly washed off under running water. The stained dish was placed under an inverted microscope, and the number of cell clones was observed to calculate the rate of clone formation = number of clones formed/number of cells seeded.
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