fluorescence microscope nikon eclipse te 2000u  (Nikon)

Journal: Scientific Reports

Article Title: Cell culture media dependent in vitro dynamics and culture characteristics of adult caprine dermal fibroblast cells

doi: 10.1038/s41598-023-38634-4

Figure Lengend Snippet: Representative pictures of chemo-attractant-driven cell migration/transwell assay. ( a ) Caprine adult dermal fibroblast cells (cadFibroblast; approximately 1 × 10 6 cells/well) in different media (100 μl) were placed in the insert’s top chambers and respective media (600 μl) containing 10 ng/ml epidermal growth factor were dispensed to the respective lower chambers of wells as the chemotactic factor. Cells were allowed to migrate for 48 h. After migration, staining of cells was performed with crystal violet, and the number of migrated cells (purple-stained) was counted in ten randomly selected fields using a bright field microscope with a 4 × objective (40× total magnification). Arrows indicate migrated cadFibroblast. ( b ) A diagrammatic representation of the transwell insert device is applied to evaluate cell migration. The small gaps between the lines represent the pores of the membrane. ( c ) Quantitative evaluation of cells moved towards the chemo-attractant (average of 10 random fields at 40 × magnification). Results are expressed as mean ± SEM. Arrowheads indicate migrated cells. Bars with dissimilar presentations (a, b, c, and d) are significantly ( p < 0.05) different. DL = DMEM with low glucose (5.5 mM), DH = DMEM with high glucose (30.0 mM), ML = α -MEM with low glucose (5.5 mM), MH = α -MEM with high glucose (30.0 mM), and FGM = Fibroblast growth medium.

Article Snippet: Migrated cells were then observed at lower magnification (4×) and were counted under a bright field microscope (Nikon Instruments, Inc., Eclipse, TE 2000U, Melville, NY, USA) in ten randomly selected fields of the fixed cells.

Techniques: Migration, Transwell Assay, Staining, Microscopy, Membrane