Axion’s Cell Plating Calculator is used to perform dilution calculations for a variety of cell plating conditions.
Included in this guide:
How to use the tool
Enter your cell count (Cell Stock Dinitial), in cells per mL, and initial cell suspension volume (Cell Stock Vinitial).
Note: An optional box allows you to enter hemocytometer values which will calculate the cell suspension density.
Enter the expected Plating Efficiency, or the % of cells that will attach. If unknown, This may be set or left at 100%. For multiple plating densities, Extra V % provides room for error in volume loss due to pipetting.
Enter your target cell plating density in the Cells per Well field, as well as the cell plating volume (Vplating) and number of wells to be plated. Multiple plating densities may be added or removed with the plus and minus buttons.
Note: Densities must be entered from highest to lowest.
You may choose to prepare aliquots using a Single Stock Method, where each aliquot is made from the same starting stock, or a Serial Dilution Method, where each aliquot is diluted from the previous aliquot.
Note: Click on the aliquot method name to change the method selected.
- Then, click Calculate Results. The following outputs are provided
Total and Needed cells provide the number of cells needed for the experiment.
- Cell Stock Vfinal provides the final volume to dilute (green alert with additional volume of media to add) or concentrate (red warning that you do not have enough cells) your sample to achieve the highest cell density chosen.
- The dilution table on the right shows how to prepare each aliquot, including how much stock or aliquot is required, how much needs to be diluted, and the volume of the final aliquot.
Definitions
Starting Conditions
| Cell Vinitial | Initial cell suspension volume. |
| Cell Dinitial | Initial cell density. Can be entered by the user or calculated from hemocytometer. |
Hemocytometer:
| Total Cells | Total viable cells counted on all squares. |
| #Squares | Number of large (4x4) squares counted. |
| Dilution | Cell:Trypan Blue dilution factor. |
Target Conditions
| Plating Efficiency | Percentage of cells attached/cells added. |
| Extra V | Percentage of overfill each aliquot will get. |
| Single Stock Method | Each aliquot is made from the same starting stock. |
| Serial Dilution Method | Each aliquot is diluted from the previous aliquot. |
| Cells per well | Target cells per well. |
| Vplating | Target plating volume (µl). |
| # Wells | How many wells to plate at this density. |
Results
| Total cells | Total starting cells. |
| Needed cells | Total cells needed to plate all wells at specified densities. Factors in extra V% and plating efficiency. |
| Cell Stock Vfinal | Final cell stock volume to dilute or concentrate to. Tip: If Total Cells >> Needed Cells, reduce Vinitial to conserve media. |
| Cell Stock Dfinal | Final cell stock density. |
Equations
