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Calculating CFU from dilution plating results

 How does a count on a plates get converted to CFUs per gram or ml


of sample? Let's illustrate the procedure with an example. Imagine
that we perform the following experiment:

 Five ml of milk are added to 45 ml of sterile broth. From this


suspension, two serial, 1/100 dilutions are made, and 0.1 ml is plated
onto Plate Count Agar from the last dilution. After incubation, 137
colonies are counted on the plate.

 This problem may be illustrated as follows:


The initial dilution is calculated
As follows:

So the initial dilution is


1/10 or 0.1 or 10-1.
 Remember, there are many ways to make 1/10 and 1/100 dilutions.

 A 0.1 ml to 0.9 ml dilution is the same as a 1 ml to 9 ml dilution and a 13


ml to 117 ml dilution.

 Next, 1 ml of the first dilution is added to 99 ml to make the second


dilution, that is a 1/100 dilution.

 This is repeated with third dilution giving another 1/100 dilution.

 Then 0.1 ml of the third dilution is plated out on a sterile plate.

The total dilution may be calculated mathematically as follows:

 To obtain the concentration of bacteria in the original sample the “dilution


factor” must be determined and then multiplied by the plate colony count.
 The dilution factor is the inverse of the total dilution –

Therefore, the “Dilution factor for our example is: 106 and the
Total Colony Forming Units (CFUs) for this sample is:

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