U cmad3 phase contrast microscope

A linear scratch on confluent MC3T3-E1 cells was performed by a sterile pipette. We removed the cellular debris by washing with PBS, and then we incubated the cells in DMEM 1% FBS alone or in the presence of various treatments for 24 h (see [41 (link)] for reference). We took the photographs immediately after the scratch (t0) and at various times after treatment (t18, t24, and t48) using an Olympus U-CMAD3 phase-contrast microscope equipped with a Zeiss Axiocam Icc1 camera at a 4× magnification. We analyzed the images using ImageJ software (NIH, Bethesda, MD, USA) as previously reported [42 (link)]. The covered area percentage was calculated for each experimental group by measuring the wound size at different times from treatment compared with the initial (t0) wound size considered as 100%.

A sterile pipette tip was used to generate a linear scratch on confluent MC3T3-E1 cells. After the remotion of cellular debris by washing with PBS, the cells were incubated in DMEM 1% FBS alone or at the presence of various treatments for 24 h. Photographs were taken immediately after the scratch was performed (t0) and at various times after treatment (t18 and t24) using an Olympus U-CMAD3 phase-contrast microscope equipped with a Zeiss Axiocam ICc1 camera at 4× magnification. The images were analyzed using ImageJ software (National Institute of Health, Bethesda, MD, USA) as previously reported [48 (link)]. For each experimental group, the percentage of the covered area was calculated by measuring the wound size at different times from treatment (t18 and t24 h) compared with the initial (t0) wound size considered as 100%.