Cells (2000 per well) were placed in 6-cm culture dishes in triplicate and cultured for 2 weeks. The colonies were fixed with 4% paraformaldehyde for 5 min, and then stained with Giemsa solution (KeyGEN BioTech, Nanjing, China) for 15 min after being washed twice with PBS. The number of colonies in each well was counted.
Giemsa solution
Manufactured by Keygen Biotech
Sourced in China
Giemsa solution is a staining reagent commonly used in the field of biology and medical diagnostics. Its core function is to selectively stain cellular components, such as nuclei and cytoplasm, for enhanced visibility and analysis under a microscope. The solution contains a mixture of methylene blue, eosin, and azure dyes that bind to different cellular structures, allowing for the differentiation of cell types and the identification of abnormalities.
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Lab products found in correlation
4 protocols using giemsa solution
1
Colony Forming Assay Protocol
2
Clonogenic Assay for Cell Viability
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HeLa or SiHa cells were seeded in 6-well plates at a density of 1,000 cells/well. Cells were then cultured at 37°C for 10 days until visible colonies appeared. Colonies were stained with 500 µl Giemsa solution (Nanjing KeyGen Biotech Co., Ltd.) and incubated for 30 min at 37°C. Colonies were then imaged using a X71 (U-RFL-T) fluorescence microscope (Olympus Corporation; magnification, ×40).
3
Colony Formation Assay Protocol
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Briefly, cells in each group were seeded in 35-mm dishes at 500 cells/dish and incubated at 37 °C in 5% CO2 for 2 weeks. The cells were fixed and stained with Giemsa solution (KeyGEN BioTECH, Jiangsu, China). Colonies consisting of at least 50 cells were counted and photographed under a microscope.
4
Clonogenic and Soft Agar Assays
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A total of 2 × 103 cells were seeded on six-well plates and maintained in a medium containing FBS for 10–14 days until visible clones appeared. For staining of colonies, 500 μl of Giemsa solution (Keygen, Nanjing, China) was added into each well and incubated for 30 min and removed followed by three washes using PBS.
For performing colony formation in soft agar, each well of a six-well plate that contained 2 ml of 0.5% (w/v) low-melting agar (Sigma–Aldrich, St. Louis, MO, United States) in DMEM medium with 10% FBS was laid in each well. Suspended cells were mixed equally, and 5 × 103 cells in 2 ml of 0.3% low-melting agar in 10% FBS were added above the polymerized base solution. Plates were incubated (37°C, 5% CO2) for 14 days before colony number, and diameter was quantified microscopically.
For performing colony formation in soft agar, each well of a six-well plate that contained 2 ml of 0.5% (w/v) low-melting agar (Sigma–Aldrich, St. Louis, MO, United States) in DMEM medium with 10% FBS was laid in each well. Suspended cells were mixed equally, and 5 × 103 cells in 2 ml of 0.3% low-melting agar in 10% FBS were added above the polymerized base solution. Plates were incubated (37°C, 5% CO2) for 14 days before colony number, and diameter was quantified microscopically.
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