Cells were stained with crystal violet. (C) Morphology of ES cells, iPS cells (iPS-MEF24, clone 1-9), and MEFs. Scale bars = 200 μm. (D) Growth curves of ES cells, iPS cells (iPS-MEF24, clones 2-1–4), and MEFs. 3 × 105 cells were passaged every 3 days into each well of six-well plates.

The cells were stained with anti-α-smooth muscle actin monoclonal antibody (N1584, Dako), anti-α-fetoprotein polyclonal antibody (N1501, Dako) or anti-βIII tubulin monoclonal antibody (CBL412, Abcam) along with 4′-6-diamidino-2-phenylindole (Sigma). Total RNA derived from plated embryoid bodies on day 6 was used for RT-PCR analysis.

4 天之前 · The expansion of the individual T cell clones directly correlated with ... in 100mL of 30% EtOH) was added and incubated at room temperature for 30 minutes. Finally, plates were rinsed to remove excess crystal violet, allowed to dry and plaques were counted to calculate viral stock titers. ... Enriched CD8 + T cells were then stained and the ...

Colonies were stained with 0.1% crystal violet (Sigma-Aldrich) and the number and type of colonies counted. Total CFE and the CFE of each colony type was calculated as a percentage of the number of cells seeded. Using a graticule and eye piece, the colony was measured across perpendicular axes to estimate the area of each colony.

Crystal Violet (CV) Staining of Cells and Clone counting Timothy Lane Stocks. 3.7% Paraformaldehyde (PFA) or 10% Formalin 0.05% Crystal Violet in Distilled Water (Filter at 0.45um before use) Fix the cells for 5 min. with 3.7% PFA STAIN them 30 minutes with 0.05%CV Wash 2x with tap water, Drain them inverted for a couple minutes.

A typical clonogenic survival experiment using adherent cells lines involves three distinct components, 1) treatment of the cell monolayer in tissue culture flasks, 2) preparation of single cell suspensions and plating an appropriate number of cells in petri dishes and 3) fixing and staining colonies following a relevant incubation period ...

2024年9月12日 · Single-cell clones were selected and cultured for 21 days. Genomic DNA extraction was performed using the column animal genomic DNA purification kit (B518251; Sangon Biotech). Growing colonies were examined using PCR amplifications for …

2025年2月21日 · Crystal violet cell viability assays were carried out ... (Sigma, P9620) at 10 µg/ml concentration and individual clones were obtained by performing serial dilution on the surviving clones. ... After removing the media and washing with PBS, cells were stained with 0.1% crystal violet (Sigma, C6158) for 15 min, washed with distilled water and ...

4 天之前 · The culture medium was refreshed every 2–3 days, monitoring for clone formation. Once formed, cells were fixed with 4% tissue cell fixative for 30 min, stained with 0.1% crystal violet for 20 ...

2019年3月26日 · Owing to the heterogeneity of mutations, it is necessary to purify cell clones grown from high dilution to the point of colony formation, which can be a time-consuming process. Here, we tested a modified approach in which we seeded cells at high dilution, together with non-edited carrier cells.