Serial Dilution
Protocol
by Nooralhuda Aljawhar
Methods of Serial Dilution
1. n-fold serial dilution:
diluting a solution by a factor of n with each step
Geometric Dilution.2
mixing equal volumes of two solutions with different
concentrations
3. Logarithmic dilution(a tenfold
dilution)
diluting a solution by a factor of the logarithm of the dilution
factor
Serial dilution calculations
In serology the process of dilution is used to quantitate the
concentration
of either Ab or Ag.; depending on this equation:Transfer volume(serum, specimen)
Concentration =
Final volume [Transfer volume + Diluent volume]
dilution factor = (final) volume/ transfer volume.
Final volume – transfer volume = diluent volume
Final volume / dilution factor = transfer volume
To calculate a compound dilution problem, the first step is to
plan the number and sizes of simple dilutions necessary to
reach the desired end point. To use the preceding example,
a1:500 dilution can be achieved by making a 1:5 dilution of
the original serum, a 1:10 dilution from the first dilution, and
another 1:10 dilution. This can be shown as follows:
Serum:
1:5 dilution 1:10 dilution 1:10
dilution
0.1 mL serum 0.1 mL of1:5 dilution 0.1 mL of 1:10
dilution
0.4 mL diluent 0.9 mL diluent 0.9 mL
diluent
Multiplying 5 * 10 * 10 equals 500, or the total dilution.
1. A 1:5 dilution of patient serum is necessary to
run a serological test. There is 0.1 mL of serum
that can be used.
What amount of diluent is necessary to make this
dilution using all of the serum?
2. Instructions that come with a buffer indicate
that it must be mixed with 19 parts of water for
use in a serological test. The volume of the buffer
concentrate is 50 mL. How would we find out the
amount of water to add and what would be the
final dilution factor?
3.If 0.02 mL of serum is diluted with 0.08 mL of
diluent, what dilution of serum does this represent?
a. 1:4
b. 1:5
c. 1:10
d. 1:20
4.If 0.4 mL of serum is mixed with 15.6 mL of
diluent, what dilution of serum does this
represent?
a. 1:4
b. 1:40
c. 2:70
d. 1:80
5.A tube containing a 1:40 dilution is accidently dropped. A 1:2
dilution of the specimen is still available. A volume of 4 mL is
needed to run the test. How much of the 1:2 dilution is needed to
remake 4 mL of a 1:40 dilution?
a) 0.2 ml
b) 0.4 ml
c) 0.5 ml
d) 1.0 ml
6.What is the final dilution of serum obtained
from the following serial dilutions: 1:4, 1:4, 1:4,
1:4, 1:4, 1:4?
7.The following dilutions were set up
to titer an antibody. The following
results were obtained: 1:4 +,1:8 +, 1:16
+, 1:32 +, 1:64 –. How should the titer
be reported out?
8.How much diluent needs to be added to
0.1 mL of serum to make a 1:15 dilution?
9. A 1:750 dilution of serum is needed to perform
a serological test. Which of the following series of
dilutions would be correct to use in this situation?
a. 1:5, 1:15, 1:10
b. 1:5, 1:10, 1:5
c. 1:15, 1:10, 1:3
d. 1:15, 1:3, 1:5
Thank you
