How to Calculate CFU From Dilution

Written by eric moll
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How to Calculate CFU From Dilution
Use a sterile metal loop to streak the sample across the surface of the agar. (Bacteria Colonies image by ggw from Fotolia.com)

CFU stands for Colony Forming Units, a term in microbiology used to quantify how many bacteria are present in a solution. Depending on the concentration of your sample, you will need to perform multiple dilutions and plate the different samples onto petri dishes. If there are too many bacterial colonies, they will be hard to count, and if there are too few, the sample may not be representative. It is generally a good idea to plate the original solution, then a 1/10 dilution (1 part solution, 9 parts saline), a 1/100 dilution and possibly a 1/1000 dilution.

Skill level:
Moderate

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Instructions

  1. 1

    Perform a preliminary count of each dish once the bacteria have had time to incubate, usually one or two days. You want to count only individual colonies, which should be distinct, isolated dots, not a whole blob of different colonies grown together. Choose the plate which has more than 30 of these colonies but less than 300.

  2. 2

    Count the number of individual colonies. This is the CFU number of your dilution -- you will have to perform a simple calculation to determine the CFU of the original sample. For this example, a hypothetical plate containing 46 colonies will be used.

  3. 3

    Determine the size of the dilution you used. Ideally, you labelled the petri dishes ahead of time. For this example, you can mix 1ml of bacterial culture with 99ml of saline. This is a 1/100 dilution.

  4. 4

    Multiply the degree of the dilution by the amount you actually plated. If, in this example, you plated 0.1ml of your 1/100 dilution onto the agar, you multiply 0.1 x 1/100, for a result of 1/1000 or 0.001.

  5. 5

    Divide the CFU of the dilution (the number of colonies you counted) by the result from step 4. For this example, you would divide 46 by 1/1000, which is the same as multiplying by 1,000. The result is 46,000 CFU in the original sample.

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