A Calculator for Cell Culture Plating

Let's take the guesswork out of cell culture.

I've created a calculator which aims to make plating cells more reproducible and easier.

Say I have 2 different cell lines which grow at different rates, and one cell line is larger than the other. I want to plate my cells into 6-well plates so they'll be 70% confluent in 4 days. How many cells of each do I plate? It's a simple question, but not a very simple answer.

This calculator will tell you, based on the numbers you know, how much media and cell suspension you should plate for an experiment in any vessel, for any length of time. You can freely change the type of plate, the timepoint, anything you want, and it will automatically calculate your resultant dilution needed. It's calculating off of confluence of the initial flask so it takes cell size and how cells grow into account as well.

This is the calculator:

Input values are in red on the left-hand side. In this scenario, I have cells which double every 53h, and I'm splitting a 90% confluent T75 flask 1:10, and resuspending the cells in 5 mL media. I count 2 million cells per mL, and I want to plate 9 wells of a 6-well plate with 2 mL media + cells. I want the cells to be 50% confluent in 80 hours, and I want 10% surplus for pipetting error.

On the right hand side, the grey highlighted cells are the answer. The calculator tells me I have enough cells for my assay (the green yes will change to a red no if not). It then tells me I need to put ~1373 uL of cell suspension into ~18.29 mL media, and plate 2 mL per well. That's it, done! Come back in 80h and they'll be at 50% confluence.

Say I decide I want to expand my assay time to 90h, and I instead want to plate my cells into a 24-well plate, all I have to do is adjust the surface area and time and it'll adjust the calculations for me.

Also included in the calculator is a reference table with surface areas for all commonly used vessels, along with the volume of media. Changing from one to another is as easy as altering the surface area and media volume.

Below this is a reference table to be filled out with your cell lines and their doubling times. This calculator makes cell culture one-stop shopping.

The calculator can be found here. Download it to your computer to edit:

https://docs.google.com/spreadsheets/d/1nxIdTN1-tx0UPDYWZzuor2wcQdKMhp6I/edit?usp=sharing&ouid=111441827851825405038&rtpof=true&sd=true

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A Calculator for Cell Culture Plating