Eclipse ts100 optical microscope

A simple scratch was created in cell monolayer and the capacity of the cells to migrate in the presence of different eugenol concentrations was studied. Images were taken with the Nikon ECLIPSE TS100 optical microscope at two time points: the beginning of the experiment then after 24 h. The microscope magnification was set at × 10. At the end point, cells were stained with crystal violet as previously described to obtain better quality photos under the microscope. Scratch diameter was measured using the Lumenera infinity analyze 6.5.5 software to quantify the scratch healing process. Three to four biological replicates were carried out.

The instant viability and death of cells were determined by trypan blue exclusion test. 100 μL of cell suspension was taken and mixed with an equal volume of 0.4% Trypan blue solution (Sigma Aldrich, St Louis, MO, USA) and allowed to stand for 5 min at room temperature. The numbers of live and dead cells were counted by ECLIPSE TS100 optical microscope (Nikon, Tokyo, Japan) with a hemocytometer and quantified by the instant cell viability rate (%): (the number of intact cells/the number of total cells) × 100.
Long-term cell viability was measured using MTT cell proliferation assays. After various treatments, cells were re-incubated in 96-well plates for 2, 8, 14, and 20 h using a method described earlier25 (link).

Hep3B and HepJ5 cells were plated onto 96-well plates, with 5 × 10³ cells per well, and cultured overnight before treatment. To evaluate the antitumor effects of AE-SN, the cells were treated with 0 to 2.0 mg/mL of AE-SN for 48 h. To evaluate the antitumor effects of integrated treatment with the chemotherapeutic drugs and AE-SN, the cells were treated with 0 to 20 μM cisplatin or 0 to 10 μM and 0, 0.5, or 1.0 mg/mL of AE-SN for 48 h. In this study, cell viability was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Table 1).
Two approaches, namely, microscopic observation and measurement of the cell size distribution, were performed to inspect the morphological changes in AE-SN-treated HCC cells. General morphological changes were observed using a Nikon Eclipse TS100 optical microscope (Nikon Instruments, Melville, NY, USA) and photographed at 100x magnification, whereas the distribution of cell diameter was measured using a Scepter cell counter (Merck Millipore Billerica, MA, USA), which divided surviving cells from cell fragments and debris in a borderline of 12 μm [18 (link)].