phase contrast microscope nikon te  (Nikon)

Journal: Current Issues in Molecular Biology

Article Title: Promising Support Coming from Nature: Antioxidant and Anti-Inflammatory Potential of Castanea sativa Wood Distillate on Skin Cells

doi: 10.3390/cimb46090556

Figure Lengend Snippet: Effect of WD on endothelial paracellular permeability in HUVEC and integrity of tight junctions affected by inflammatory damage. ( A ) The permeability assay was performed on HUVEC seeded on transwell inserts. The confluent monolayers were treated with WD [0.04–0.07%, ( v / v )] and an inflammatory mix (MIX: IL-1β [100 ng/mL] + TNF-α [10 ng/mL]). At the end of the treatment, FITC–dextran (10 µM) was added and permeability in the HUVEC monolayer was detected as passage of FITC–dextran from the upper to the lower compartment of the transwell insert. The extent of permeability was determined by measuring the fluorescence at 485/535 nm, excitation/emission, respectively, after 15 min. Data are reported as fluorescence unit (n = 3) ± SD; *** p < 0.001 vs. untreated cells (CTR); ### p < 0.001, compared to the MIX. From a molecular point of view VE-cadherin ( B ) and ZO-1 ( C ) were assessed by immunofluorescence analysis of HUVEC treated for 18 h with WD [0.04–0.07%, ( v / v )], alone or in combination with the inflammatory mix (MIX: IL-1β [100 ng/mL] + TNF-α [10 ng/mL]. VE-cadherin fluorescence was captured by a secondary antibody conjugated with Alexa Fluor 555 ( B ). ZO-1 was visualized by Alexa Fluor 488 ( C ). DAPI-stained nuclei in blue. Images were obtained by a Nikon Eclipse TE 300 microscope (magnification 60×). The merge images were obtained using FIJI ImageJ software 6.1.1.jar. The presented pictures are representative of n = 3 experiments.

Article Snippet: Images of the wound in each well were acquired from 0 to 18 hours using a phase contrast microscope (Nikon Eclipse TE 300, Nikon, Tokyo, Japan), at 10× magnification.

Techniques: Permeability, Fluorescence, Immunofluorescence, Staining, Microscopy, Software

Journal: Poultry Science

Article Title: Exosomal circ_CCDC7/gga-miR-6568-3p/Pax7 axis accelerates the differentiation of chicken embryonic stem cells infected with subgroup J avian leukosis virus

doi: 10.1016/j.psj.2024.103898

Figure Lengend Snippet: Effect of ALV-J on ESCs. (A and B) The morphology of ESCs infected (cESC-ALV-J) or uninfected ( cESC ) with ALV-J under an phase-contrast microscope at 20X (A) and 40X (B). (C) Cell proliferation and cell number calculation. (D) RNA concentration of exosomes from ESCs infected or uninfected with ALV-J. *, P < 0.05. **, P < 0.01. ***, P < 0.001. The scale bar indicates 250 μm. (E) Western blot analysis on cESC-ALV-J and cESC. Results use antibodies against skeletal muscle-specific markers (MYHC and Pax7) and cESC markers (Oct4 and Nanog).

Article Snippet: Subsequently, the filter paper was picked up with forceps, and the cESCs in the embryonic disc adsorbed on the filter paper were rinsed into the PBS in the 1.5 mL eppendorf tube. cESCs in PBS were centrifuged at 1,000 rpm for 5 min, resuspended in Dulbecco's Modified Eagle Medium ( DMEM ), seeded at a density of 1 × 10 5 cells in a 6-well plate, and cultured at 39°C with 5% CO 2 for 72 h. The DMEM included high glucose medium with 10% fetal bovine serum ( FBS ; Thermo Fisher Scientific, Inc., Waltham, MA), 2% chicken serum, 1% sodium pyruvate, 1% glutamax, 1% non-essential amino acids ( NEAA ; Thermo Fisher Scientific, Inc., Waltham, MA), 10 μg/mL leukocyte inhibitory factor ( LIF ; Thermo Fisher Scientific, Inc., Waltham, MA), 10 μg/mL stem cell growth factor ( SCF ; Thermo Fisher Scientific, Inc., Waltham, MA), and 50 μg/mL fibroblast growth factor 2 ( FGF2 ; Thermo Fisher Scientific, Inc., Waltham, MA). cESCs’ cultural medium was changed every 2 d, and passaging was done every 3 d. Cells were visualized under a bright-field phase-contrast microscope (Nikon-Eclipse TE 2000-S).

Techniques: Infection, Microscopy, Concentration Assay, Western Blot