Microbiology Dilutions

Theory and Practice

What are they for?

For many years, scientists have used a serial dilution method to control sample size. This is important to many diagnostic and enumeration methods.

Key Terms
Sample: refers to the original liquid that will be tested. If a rinse of a chicken skin is being tested, what is the sample? The liquid from the rinse is
the sample.

Diluent: the solution that is mixed with the sample (often purified water). Concentration: refers to how much of the sample is in a given test. For example, 40 to 1 (1/40) dilution means that for every 40 parts of water, there is 1 part of sample. Dilutions are given in terms of concentration.

Pitcher of Juice
For example, this is 100% pure juice. Or in other words, this is a one to one dilution.

So the concentration is 1 to 1, or 1/1.

Concentra on Volume
Even though it is made up of many ingredients, juice is our sample. And even if we were to pour it into a glass, its still 100% juice in that container.

Still 1/1 Dilution.

Serial Dilution
2.) If we take 10 ml of that sample and put it in an empty tube, it is still the same concentration. Dilution = 1/1 1.) Serial Dilution starts with a primary sample. Dilution = 1/1

Serial Dilution

But for dilutions, these other tubes are filled with 9 ml of water.

Serial Dilution
1 ml

We then transfer 1 ml of the original solution into the tube with 9ml of water. Tube #2 now has 10 ml of liquid. #1 #2 #3
1 ml SAMPLE #4

9 ml H2O #5

= 10 ml

Serial Dilution
1 ml

#1

#2

#3

#4

#5

We then transfer 1 ml of the original solution into the tube with 9ml of water. Tube #2 now has 10 ml of liquid.
1 ml SAMPLE

9 ml H2O

= 10 ml

Serial Dilution
Now, tube #1 is still completely the original concentration. (1/1 Dilution)

#1

#2

#3

#4

#5

Tube #2 has 10 ml, of which 1 ml is sample. (1/10 Dilution)

Serial Dilution
1 ml

#1

#2

#3

#4

#5

1 ml is then taken from tube #2, and mixed into tube #3

Serial Dilution
1 ml

#1 1/1

#2 1/10

#3

#4

#5

1 ml is then taken from tube #2, and mixed into tube #3 Tube #3 now has 9 ml of water, and 1 ml from tube #2 9 + 1 = 10 ml

Test Yourself
What is the concentration in tube #3?

# 1 1/1

# 2
1/10

# 3

# 4

# 5

Lets Work It Out

To find the concentration of tube #3, we multiply:

1 10
the previous concentration (tube #2)

1 10

Units transferred (ml)

over

Total units in new container

Lets Work It Out

To find the concentration of tube #3, we multiply:

1 x 10
the previous concentration (tube #2)

1 10

Units transferred (ml) over

1 100

Total units in new container

So if we continue the process

#1 1/1

#2 1/10

#3 1/100

#4 1/1,000

#5

So if we continue the process

#1 1/1

#2 1/10

#3 1/100

#4 1/1,000

#5 1/10,000

So if we continue the process

#1 1/1

#2 1/10

#3 1/100

#4 1/1,000

#5 1/10,000

Consistent Formula

#1 1/1

#2 1/10

#3 1/100 Units transferred

#4 1/1,000

#5 1/10,000

Previous concentration

x Total units

New Concentration

But what happens when we mess everything up?

TEMPO Problems

TEMPO vials = 4 ml

Find the Final Dilution:

Find the Final Dilution:

1 1 x 1 4 1 = 4

1 100

1 4

1 = 400

Always Remember
1.) Concentration Volume

2.) Follow the formula

Units transferred Previous concentration x Total units = New Concentration