I want to use, 1:10 MOI, (cell: bacteria).
Assuming that one well of cell is 1 x 10^5, thus, the no of bacteria per well should be 10 x 10^5.
The bacteria is V. cholerae, and from my reading, at A600 = 1.0, correspond to 5 x 10^8 CFU/ml.
Therefore, I should find the OD of bacteria that correspond to 10 x 10^5.
After obtaining the OD, I use dilution formula, and from that, I took 10ul and infect each well. As a result, each well will be infected with 10 x 10^5 bacteria.
Am I right? Thank you.
The method I used is as such:
obtain an OD-CFU standard curve based on OD measurement AND agar plating (usually, plate 100uL of diluted suspension; the suitable dilution factor is 5x10^5).
Then, prepare a culture (e.g. log phase), wash with buffer and concentrate it at least 10x and normalize its concentration to 10^10/mL or 10^9/mL.
When you add 1uL of the above to a 1mL final volume (e.g. with your host cells), you dilute the original concentrations (10^10/mL or 10^9/mL) by a factor of 1000x to become the working concentration of 10^7/mL or 10^6/mL.
To summarize, all you need to do is to prepare a stock solution for 1000x dilution. The unit of the bacteria density should be xxx bacti/mL (not xxx bacti/uL).
Be very careful that when you concentrate the bacteria (esp. V. cholerae), their density becomes more dangerous and prone to infect humans.
The method was described in our paper (supporting info), attached.
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