Ts2 s sm inverted microscope

HCT116 and HT29 cells in the logarithmic phase were used after trypsin digestion and inoculated into a 6-well plate at a density of 6 × 10⁵ cells per well of 2 ml. The cell fusion rate was close to 100% after 24 h of incubation. A 20-μl pipette tip was used to make vertical scratches on the bottom of the 6-well plate (3 strips per well). Then, the plate was washed three times with PBS solution. Following that, a serum-freedrug-containing medium was added for continued incubation. Finally, images were collected at the appropriate time using a Nikon TS2-S-SM inverted microscope and then processed using ImageJ software (version 8.0, Madison, WI, USA) to calculate the proportion of cell-free area in the treated group versus the control groups [16 (link), 17 (link)].

During the logarithmic growth stage, HCT116 and HT29 cells were inoculated in one 6-well plate (1 × 10⁶ cells per well). The cells were transfected when the growth density was 40-50%, and replaced with complete culture medium after 6 h. When the cell confluence was approximately 90%, a 10 μl pipette tip was adopted with a view to making the vertical scratches on the bottom of that plate. Next, that plate was rinsed 3 times using PBS solution. Then, 2 mL of serum-free, fresh medium was placed for continuous hatching. Photographs were taken 48 h later via one Nikon TS2-S-SM inverted microscope and handled by means of ImageJ software to calculate the healing rates of cells in both groups, where the healing ratio (%) = (scratch spacing at 0 h - scratch spacing at 24/48 h) / scratch spacing at 0 h × 100%.