Infinity analyze software

GBM neurospheres were dispersed by Accutase (AT‐104) into single cells, and 10⁴ cells were seeded in triplicate into six‐well plates in Neurobasal culture medium. At indicated time points, neurosphere formation was measured by infinity analyze software (Lumenera Corporation, Ottawa, ON, Canada) under Olympus microscope (Tokyo, Japan).

Calcein-AM (Life Technologies, Carlsbad, CA, USA) was used to determine the cell viability in accordance with the manufacturer’s protocol. Specifically, hiPSC-RPE cultures were incubated in the dark with Calcein-AM (5 µM) and nuclear staining dye (Hoechst, 1:2000; Life Technologies) for 30 min at 37 °C. Calcein-AM and Hoechst-stained hiPSC-RPE samples were imaged on a Leica DM IRBE inverted microscope (Leica Microsystems, Buffalo Grove, IL, USA) using a Lumenera INFINITY3–1 camera and INFINITY ANALYZE software (Lumenera, Ontario, Canada).

The assays evaluate the antiviral activity of the compound when administered before, during or after HSV-2 infection. Cells were incubated with different concentrations of A-3302-B (from 100 μM to 0.4 μM) in a 24-well plate at 37 °C for 2 h before infection (pretreatment), for 2 h during infection (cotreatment) or 24 h after removal of virus inoculum (post-treatment). Cells were concurrently infected with HSV-2 at MOI of 0.001 PFU/cell and treated for plaque reduction assay. Plaque size was measured with an Axiovert 200 inverted microscope (Zeiss, Jena, Germany) equipped with Infinity3 microscope camera (Lumenera, Ottawa, ON, Canada) and Infinity Capture software (Lumenera), and was analyzed with Infinity Analyze software (Lumenera) and ImageJ software (Bethesda, MD, USA).

Image samples were chosen to have distinct grains entirely in focus. Ceria pellet samples from previously published work were used [37 ]. Briefly, the samples were synthesized from 99.9% cerium oxide powder sieved to 212 µm particles. After pressing, the pellets were sintered at 1650 °C for 2 h in the air before being annealed at 1750 °C for 10 h to induce grain growth and grain boundary differentiation. Images of the resulting microstructure of 8 mm sample pellets were collected using a Nikon Eclipse LV100 optical microscope with a Lumenera Infinity 1 microscope camera attachment. Images were collected as 1280 by 1024 resolution TIF files, each approximately 9 megabytes in size. Lumenera’s Infinity Analyze software was used to calibrate the scale of the image and to place a scale bar in each bottom-right corner. Five fields (images) were acquired for five samples at a magnification of 10×. The test area for each image was approximately 0.655 mm².