Next, nonadherent cells were removed, and fresh media was added until migrated cells reached 80–90% confluency, displaying a consistent spindle-shaped appearance. The desired confluency, morphology and adhesion of hGF cells were determined by using a phase-contrast inverted microscope (Carl Zeiss Axiovert 40C Imaging Microscope). Plastic-adherent confluent cells were intermittently passaged with 0.25% trypsin-EDTA (Gibco) for 2–3 min at 37 °C in gently rocking flasks. Trypsin neutralization was achieved using complete DMEM (twice the volume used for the trypsin-EDTA reagent). Cell suspensions were then centrifuged at 1500 rpm for 5 min. The next step consisted of supernatant aspiration and discarding. The cell pellets were then resuspended in a 5 mL complete medium volume to increase cell numbers. After diluting the cell pellet and dividing them into two T-75 flasks, fresh growth medium was added to expand the cells. Observations were carried out routinely under an inverted phase-contrast microscope (Fig. 1 -B). The experiments were conducted with cells from the sixth passage (Saczko et al., 2008 (link), de Abreu et al., 2019 ).
Axiovert 40c imaging microscope
Manufactured by Zeiss
Sourced in Germany
The Axiovert 40C Imaging Microscope is a versatile laboratory equipment designed for general imaging applications. It provides a stable and reliable platform for various microscopy techniques, including brightfield and phase contrast imaging. The Axiovert 40C features a compact and ergonomic design, making it suitable for use in a variety of laboratory settings.
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Lab products found in correlation
2 protocols using axiovert 40c imaging microscope
1
Expansion and Passaging of Human Gingival Fibroblasts
2
Wound Closure in PDLF Cells
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The wound closure ability of PDLFs in different glucose concentrations was measured using the scratch migration assay. PDLFs cells were plated on 12-well culture plates at a density of 5 × 104 cells/well, incubated in a complete culture medium, and grown overnight at 37 °C, 100% humidity, 95% air, and 5% CO2 to achieve confluency. A sterile ruler was used to reference the center, then a scratch was manually produced across the cell monolayer using a sterile p200 pipette tip (Thermo Fisher Scientific). Cells were then washed with PBS to remove cellular debris and exposed to each glucose concentration with/without 1 µg/mL of LPS. After scratching, the wound closure was observed at baseline, 6, 24, and 48 h under a phase-contrast microscope (Carl Zeiss Axiovert 40C Imaging Microscope, Göttingen, Germany) until the wound healed in all the groups, and digital photographs were captured by matching reference points. The percent wound closure was calculated using ImageJ software (National Institutes of Health, Bethesda, MD, USA) version 1.50i. Data are representative of 3 replicas for each experimental condition, where 3 images were captured per condition.
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