Plating 2 48 well plates of Dv-1 cells for DiNV titering
Using 2 48 well plates with the interior 20 wells only, meaning I’ll need 40 wells of cells. Plates will have this layout:
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | |
|---|---|---|---|---|---|---|---|---|
| A | cell control | 10^-1 | 10^-2 | 10^-3 | 10^-4 | 10^-5 | ||
| B | 500ul medium | 200ul 10-1 dilution | 200ul 10-2 dilution | 200ul 10-3 dilution | 200ul 10-4 dilution | 200ul 10-5 dilution | ||
| C | 500ul medium | 200ul 10-1 dilution | 200ul 10-2 dilution | 200ul 10-3 dilution | 200ul 10-4 dilution | 200ul 10-5 dilution | ||
| D | 500ul medium | 200ul 10-1 dilution | 200ul 10-2 dilution | 200ul 10-3 dilution | 200ul 10-4 dilution | 200ul 10-5 dilution | ||
| E | 500ul medium | 200ul 10-1 dilution | 200ul 10-2 dilution | 200ul 10-3 dilution | 200ul 10-4 dilution | 200ul 10-5 dilution | ||
| F |
- Scraped 1 flask of confluent Dv-1 cells and spun down at 800rpm for 4 minutes in a 15mL conical
- Removed the supernatant
- Resuspended the cells in 1mL medium
- Used 20ul in a hemocytometer to count the cells:
- Q1: 494 cells
- Q2: 578 cells
- Q3: 468 cells
- Q4: 512 cells
- Average: 513 cells
- Total in the 1mL that should be 5,130,000 cells
- I was not sure how to do the calculation here at the time, I just resuspended all the cells to a total of 10mL
- Then added 200ul of that mixture to each of the 40 wells in the plate
- this ends up being about 100,000 cells per well
- These cells were let to sit in the 23C incubator overnight before inoculation