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Dilution

There are three types of dilution: simple dilution which involves only a single dilution, serial dilution which is done in a series with the same dilution factor, and compound dilution which is similar to serial dilution but uses a different initial dilution for each tube. A serial dilution procedure is described involving labeling 10 test tubes, adding saline and antibody to the first tube and transferring 0.25ml between tubes, then adding red blood cells before incubating and reading the results to determine the endpoint and titer. Interpretation of negative and positive results and dilution factors is also briefly explained.

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184 views8 pages

Dilution

There are three types of dilution: simple dilution which involves only a single dilution, serial dilution which is done in a series with the same dilution factor, and compound dilution which is similar to serial dilution but uses a different initial dilution for each tube. A serial dilution procedure is described involving labeling 10 test tubes, adding saline and antibody to the first tube and transferring 0.25ml between tubes, then adding red blood cells before incubating and reading the results to determine the endpoint and titer. Interpretation of negative and positive results and dilution factors is also briefly explained.

Uploaded by

mir40323
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Dilution

simple dilution
Introduction Only a single dilution is made in type of
dilution
Dilution is an indication of
relative concentration which are
a form of ratio serial dilution
A dilution involves two entities: a type of dilution that is done by series
the solute, which is the material and with same dilution factor
being diluted, and the diluent,
which is the medium making up
the rest of the solution. compound dilution
there are three types of dilution: Similar to serial dilution however it have
simple dilution, serial dilution, a different initial dilution each tube
and compound dilution
Serial Dilution

Preanalytical phase Analytical phase


Material
Antibody A (anti-A antisera)
Prepare the working area by
Refrigerator
sterilizing the site.
3% Red blood cell suspension
wear proper PPE Test tube rack

Reagent must be brought to 0.85-0.90% Saline (Normal


Saline Solution/NSS )
room temperature and
shaken well before use Water proof marker

Prepare a 3% Red blood Cell Ten 12 x 75 Test Tubes


Suspension (RCS)
Three serological pipettes
Procedure

Label ten test tubes 1-10 Draw up to 0.25 ml from tube Mix well and show your
Place 0.25 ml of saline in each of #1 and transfer to tube #2. instructor your tubes.
the ten tubes. Again raise and lower to Place in refrigerator for 30
Use a clean serological pipette to solution into the pipette three minutes.
draw up 0.25 ml antibody A. Add
times to mix. Remove from refrigerator,
the antibody to tube #1 by
Repeat step 4, transferring centrifuge for 20 seconds.
carefully lowering and raising the
0.25 ml tube #2 to tube #3, Read immediately for
solution into the pipette to tube
then #4, then #5, etc. Discard agglutination by gently
#1 by carefully lowering and
0.25 ml from tube #5. shaking the tube to dislodge
raising the solution into the
Use clean serological pipette the red blood cell button.
pipette three times to mix, being
careful to avoid creating bubbles
to add 0.25 ml of 3% red
in the mixture. blood cell suspension to each
tube.
POST- ANALYTICAL PHASE

Cleaning and Disinfection


All infectious materials should be disposed
appropriately.

The working area should be cleansed with the


disinfectant before leaving.

The PPE of each individual should be removed


properly. These cannot be exposed outside
the laboratory premises.
Interpretation of Results

Negative Result Endpoint Titer Dilution Factor

Clumping of the It is used to


red blood cell It is the last tube the reciprocal of
correct for having
suspension showing the last dilution
used a diluted
agglutination showing a positive
sample in a
false negative result
determination
reactions occur
rather than the
when the tubes are
undiluted sample.
shaken too roughly
DILUTION FORMULA
𝐷𝐼𝐿𝑈𝑇𝐼𝑂𝑁 = Amount of Solute
Total Volume

TOTAL VOLUME= Amount of solute +


diluent
THANK YOU!

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