Materials and Methods
SF9 cells were gently sloughed from culture to maintain viability. Cells were centrifuged at 100 g then resuspended in PBS for counting.
Trypan Blue
Cells in PBS were mixed 1:1 with Trypan Blue (DeNovix CAT #CD-TRY-1.5) and counted using the Trypan Blue app on the CellDrop Cell Counter using the default protocol described below:
Table 1: SF9 counting protocol for the Trypan Blue app.
Dilution Factor | 2 |
Minimum Diameter | 6 |
Maximum Diameter | 30 |
Live Roundness | 50 |
Dead Roundness | 25 |
Stained Threshold | 35 |
Small Cell Mode | OFF |
Irregular Cell Mode | OFF |
AO/PI
SF9 cells in PBS were mixed 1:1 with AO/PI (DeNovix CAT #CD-AO-PI-1.5) and counted using the AO/PI app on the CellDrop FL Automated Cell Counter using the protocol outlined below:
Table 2: SF9 counting protocol for the AO/PI app.
Dilution Factor | 2 |
Minimum Diameter | 8 |
Maximum Diameter | 20 |
Live Roundness | 1 |
Dead Roundness | 1 |
Green FL Threshold | 1 |
Red FL Threshold | 12 |
Irregular Cell Mode | OFF |
Results and Summary
The CellDrop Automated Cell Counter counts cells and reports viability, circling live cells in green and dead cells in red. The Trypan Blue app identifies stained (dark) cells as dead and unstained cells as live and labels them appropriately (Figure 1). The AO/PI app identifies green-fluorescing cells as viable live cells, and red-fluorescing cells as dead (Figure 2). CellDrop enables rapid and accurate counting and identification of live and dead cells in a variety of mammalian and other cell types, including the SF9 insect cell line.
18-JUL-2022