Volocity visualization software

IHC performed with ovarian cancer tissue microarray and animal tissue specimens. For IHC, the samples were incubated with primary antibody, followed by incubation with HRP-conjugated secondary antibody. METTL14 FISH was performed to analyze the copy numbers of the METTL14 gene in ovarian cancer and normal tissue samples. Briefly, tissue sections were deparaffinized in xylene, rehydrated in a series of ethanol solutions, and then subjected to FISH with the METTL14 FISH probe (1–2.5 nM) in a buffer containing 2× saline-sodium citrate, 25% formamide, 10% dextran sulfate, and 1 mg/mL yeast in a humidified chamber for 3–4 h at 42°C, according to the manufacturer’s recommended protocol. The specific FISH signal was detected as brown punctate dots. Images were captured using a Nikon 90i microscope-equipped orca ER camera (Hamamatsu Photonics, Hamamatsu, Japan) with a 60×, 1.4 N.A. VC objective lens and were quantified using the Volocity visualization software (Perkin elmer, Waltham, MA, USA).

For imaging purposes, 6 to 10 worms were immobilized in 35 mm optical glass bottomed dishes (MatTek Corporation, Ashland, MA) with 6 µl of 0.1 mM sodium azide in PBS. Confocal images were taken using a Leica TCS SP8 microscope. GFP fluorescence was illuminated using a 488 nm argon laser line with a 63×1.4NA oil Apochromat CS2 objective. Fluorescence was captured using a spectral HyD detector over ∼100 Z-planes. Confocal images were visualized, rendered and analyzed using Volocity Visualization Software (v 5.4, PerkinElmer).