Eclipse ti s microscope

Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker ARX (300 MHz) spectrometer as deuterochloroform (CDCl₃) or DMSO‑d₆ solutions using tetramethylsilane (TMS) as an internal standard (ppm = 0) unless noted otherwise. High-performance liquid chromatography (HPLC) was performed on a Waters Alliance 2695 with a photodiode array 2996 system using a Waters XTerra® RP18 column (3.5 µm, 4.6 × 150 mm). All compounds used in anti-ovulation studies had purity than >95% at MAX plot as determined by this system. Medium-pressure liquid chromatography (MPLC) was performed using a Isolera One from Biotage USA® (Charlotte, NC) utilizing SiliaFlash® P60 40–63 µm silica gel from Silicycle (Quebec, Ontario). Thin-layer chromatography (TLC) analysis were carried out on silica gel GF (Analtech) glass plates (2.5 cm × 10 cm with 250 µm layer, pre-scored). Most chemicals and solvents were analytical grade and used without further purification unless otherwise noted. LNG was supplied as a gift from Wyeth (DE) and ENG was purchased from Nexconn (PRC). A Mini-Mill Pulverisette 23 purchased from Fritsch was used to mill the solid compounds for formulation. Female Sprague-Dawley rats were purchased from Charles River (Wilmington, MA). Clean tip sponge swabs were purchased from Texwipe (Kernsville, NC), and vaginal smears were visualized using a Nikon Eclipse Ti-S microscope.

Cells grown on coverslips were fixed with ice-cold methanol and staining performed as described previously [35 (link)]. Myc was visualised using an anti-Myc primary antibody and Alexa Fluor^® 488 secondary antibody, and the nucleus counterstained with DAPI. The cell staining was visualized using an Eclipse Ti-S microscope (Nikon).

Adherence of PMA-treated THP-1 cells was determined at the indicated time points. Cells were harvested by trypsinization and resuspended in RPMI. The cell number was determined using 0.4% trypan blue staining with a Vi-Cell XR Cell Viability Analyzer (Beckman Coulter, Brea, CA, USA). To determine the relative adherence, adherent cells were collected and percentage adherence was calculated as the number of adherent cells divided by the starting cell number and multiplied by 100.
Images of cells were collected with an Eclipse Ti-S microscope (Nikon GmbH, Tokyo, Japan) equipped with a Nikon Digital Sight DS-Fi1c color camera (Nikon GmbH) and a 20× objective (NA = 0.45, S Plan Fluor, Nikon GmbH) or a 10× objective (NA = 0.3, Plan Fluor, Nikon GmbH).

To study cell migration, a scratch assay was conducted. Cells were seeded in six‐well plates to full confluency. A 10 μL pipette tip was used to create a scratch, followed by medium replacement. Monitoring and imaging occurred at 0, 12, and 24 h using a Nikon ECLIPSE Ti‐S microscope. In addition, migration potential was assessed through a transwell migration assay. Cells were seeded in the upper chamber of a 24‐well Transwell (8 μm pore, corning) with a lower chamber containing 10% fetal bovine serum. After 24 h, nonmigrating cells were removed, and migrating cells were fixed, stained, and observed using the same microscope system, along with NIS‐Elements F v4.0 software for data collection.