Magellan 400 scanning electron microscope

Peritoneal tissues from female c57bl/6 mice were excised, rinsed twice, and submerged in PBS. While submerged, tissues were uniformly trimmed to a 2.0 × 2.0mm square and pinned to a SYLGARD® 184 silicone coated dish. Cells to be assayed were labeled with CellTracker™ Green, trypsinized, diluted to 200,000cells/ml, and 2.5ml of cell suspension was added to tissue and incubated for 2 hours at 37°C. After incubation, the tissue was washed vigorously five times, removed from the silicone bed, and mounted onto a glass coverslip for imaging (EVOS® FL) and enumeration. The number of cells/5 fields was counted and the assay was performed in triplicate. Results show mean +/− SEM. Student’s t-test was used to determine p values (Sigmaplot). Alternatively, tissues were processed for scanning electron microscopy, as described previously (Barbolina et al. 2013 (link)). Briefly, post-assay tissues were washed, fixed, dehydrated, and dried. Once dried, samples were placed on carbon stubs, subjected to Flash-Dry™ silver paint, coated with platinum, and examined using a Magellan 400 scanning electron microscope (FEI, Hillsboro, Or).