Juli br microscope

Two cell types were used in the study: primary gingival fibroblasts; normal, human, adult (HGF) from ATCC (Manassas, VA, USA); and CHO cells purchased from Sigma-Aldrich (The European Collection of Authenticated Cell Cultures—ECACC). Cell cultures were carried out in an incubator at 37 °C, in a 5% CO₂ atmosphere, at 95% humidity. Cells, after thawing, were cultured for at least 2 weeks prior to testing. During culturing, confluence measurement was performed using a Juli Br microscope (NanoEntek, Seoul, Republic of Korea). Cell cultures were passaged once a week with trypsin/EDTA solution. Cells for the assay were counted using a NucleoCounter^® NC-200 automatic cell counter (ChemoMetec A/S, Allerod, Denmark). Cells were cultured in Dulbecco’s modified eagle medium (DMEM) without phenol red or F-12K medium (Kaighn’s modification of Ham’s F-12 medium) supplemented with 10% fetal bovine serum (FBS), antibiotics, and L-glutamine (200 mM). Culture reagents were purchased from Biological Industries (Beit-Haemek, Israel). Detailed methodology for the assessment of cell vitality is presented by Hadzik et al. concerning experimental implant surfaces [16 (link)].

To assess the effect of the test compounds on tumor metastasis, a migration assay was performed. The cell culture monolayer was scratched using the SPLScar™ Scratcher cell migration assay system (Figure 2). Then, the tested oils were added to the cell cultures. In the migration assay, the lowest active concentration was selected for all tested oils. Photographs were taken with a Juli Br microscope (NanoEnTek Inc., Seoul, Republic of Korea), then the cell cultures were incubated for 24 h and the scratch was photographed again. In addition, the rate of crack overgrowth was analyzed by monitoring the culture using a Juli microscope. Using the ImageJ 1.54d open software platform, the length of the scratch was measured after its preparation, 24 h after incubation with the test compounds, and at 15 min intervals to determine the rate of overgrowth.

The Normal Human Dermal Fibroblast (NHDF) cell line from Lonza (Basel, Switzerland) and the L929 cells were purchased from Sigma-Aldrich (Merck Group, Headquarters, Darmstadt, Germany) (The European Collection of Authenticated Cell Cultures—ECACC). All cells were cultured at 37 °C, in 5% CO₂, 95% humidity in a CO₂ incubator. The cells were cultured for a minimum of 2 weeks before the assay. If the confluence exceeded 70% (confluence measurement, JuliBr microscope, NanoEntek, Seoul, Korea), the cell cultures were passaged with trypsin/EDTA solution. The cells were counted with an automatic cell counter NucleoCounter^® NC-200 (ChemoMetec A/S, Allerod, Denmark). If the confluence was less than 70%, the medium was replaced with fresh medium. The cells were cultured in Dulbecco’s modified Eagle medium (DMEM) without phenol red, 10% fetal bovine serum (FBS), penicillin (10,000 U/mL), streptomycin (10 mg/mL), and L-glutamine (200 mM). All culture reagents were purchased from Biological Industries (Beit HaEmek, Israel).