Imaging Flow Cytometry Dataset For Aged Buccal, Contact Epithelial and Vaginal Cells

Buccal and contact samples were obtained pursuant to VCU-IRB approved protocol ID#HM20000454_CR3. The ten volunteers for buccal samples were asked to swab the inside of cheek for 30 seconds. Swabs were left to dry for 1 hour to 6 days. For contact samples, ten volunteers (six of whom were buccal cell donors) were asked to hold/rub a conical tube (P/N 229421; Celltreat Scientific) for five minutes to deposit cells. Tubes were left out for 24 hours to 5 days to dry before collecting cells. Cells were collected from the surface with 1 sterile, pre-wetted swab, and 1 sterile, dry swab. Ten vaginal samples were obtained pursuant to VCU-IRB approved protocol ID#HM20002931_Ame2. Volunteers were asked to swab the inside of the vaginal cavity, and swabs were dried and stored at room temperature until analysis. Storage times ranged from 48 hours to approximately eight weeks. Swabs were eluted in 1 mL of 1x Cell Staining Buffer (P/N 420201; Biolegend), and gently vortexed for 10 seconds. Samples were centrifuged at 1500 × g at 4°C for 5 minutes. The supernatant was discarded, and the cell pellets were dissolved in 100 uL of 1x Cell Staining Buffer for imaging flow cytometry. All samples were analyzed using an Amnis® Imagestream X Mark II (EMD Millipore, Burlington MA) equipped with 405nm, 488nm, 561nm, and 642nm lasers. Laser voltages for all tests were set at 120mW, 100mW, 100mW and 150mW, respectively. Images of individual events were captured in five detector channels labeled: 1 (430-505nm), 2 (505-560nm), 3 (560-595nm), 5 (640-745nm), and 6 (745-780nm). Channel 4 was used to capture Brightfield images. Magnification was set at 40x and autofocus was enabled so that the focus varied with cell size.