Clonogenic Assay for Cell Survival

A total of 2 × 10³ AMC-HN-8- and TU177-transfected cells were seeded into the respective well of a six-well plate and incubated at 37°C in a humidified incubator containing 5% CO₂. Ten days later, the cell colonies were rinsed gently with PBS in triplicate, fixed with 4% paraformaldehyde for 20 min, and then stained with 0.5% crystal violet for 20 min. Lastly, the number of colonies formed with more than 50 cells per well was calculated using a microscope (CKX53, Olympus Corp.) at 200× magnification.

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Yu L., Cao H., Yang J.W., Meng W.X., Yang C., Wang J.T., Yu M.M, & Wang B.S. (2023). HDAC5-mediated PRAME regulates the proliferation, migration, invasion, and EMT of laryngeal squamous cell carcinoma via the PI3K/AKT/mTOR signaling pathway. Open Medicine, 18(1), 20230665.

Publication 2023

CKX53

A 103 Cells Crystal violet Microscope Paraformaldehyde

Corresponding Organization : Second Hospital of Hebei Medical University

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Variable analysis

independent variables

Cell type (AMC-HN-8- and TU177-transfected cells)

dependent variables

Number of colonies formed with more than 50 cells per well

control variables

Incubation temperature (37°C)
Incubator CO2 concentration (5%)
Incubation duration (10 days)
Cell seeding density (2 × 10^3 cells per well)
PBS rinse (in triplicate)
Fixation with 4% paraformaldehyde (20 min)
Staining with 0.5% crystal violet (20 min)
Microscope magnification (200×)

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