LAB – WEEK 12: Blood Composition, Types + Compatibility
Biology I, Pre-Health Sciences – Durham College, 2023-2024
INSTRUCTIONS
Before coming to the lab, familiarize yourself with the
goals, agenda, and required prior learning.
Goals:
Goals:
• Building Fake Blood
o List the components of blood plasma
and the various cell types present within
using correct terminology
o Follow a scientific protocol
o Demonstrate ability to transfer small
volumes of liquids with P20, P200,
P1000 micropipettes using proper
technique
o Accurately weigh a specific reagent
using a standard balance
o Demonstrate proper use of various lab
equipment/reagents (E.g. centrifuge)
• Blood Typing and Compatibility
o Describe the biology of ABO/Rh blood
types using correct terminology
o Describe using correct terminology why
a blood transfusion is safe or unsafe
given various ABO/Rh blood types
o Follow a scientific protocol; Collect
observations and record data; Analyze
data using deductive reasoning to form
conclusions; Compare experimental
data to positive and negative controls.
o Demonstrate proper use of various lab
equipment/reagents (E.g. ABO/Rh blood
and antibodies)
Assessment Policies:
• During the lab you will be asked to demonstrate your knowledge and
understanding of this week’s topics via formal and informal assessments.
These assessments are closed-book. They must be submitted in-class during
your scheduled lab period at the discretion of your instructor. No late
submissions are accepted. EXCEPTION: ASC students with specific
accommodations may be allowed to submit their evaluations after the
scheduled lab period.
Come Prepared:
• To be successful in this lab, you will need to complete the following before
attending the lab:
• Review this lab booklet thoroughly, including all links, appendices, etc.
• Bring your own lab coat and safety glasses
• Learn all online content from DC Connect pertaining to Week 11 >
Cardiovascular System – Part 1 > Blood Composition and Blood
Compatibility. You should be able to:
• Describe how blood separates when spun in a centrifuge
• List the major solutes and plasma proteins found in blood plasma
• List the scientific names for red blood cells, white blood cells, and
platelets
• Describe the basic structure of a red blood cell and explain how
this structure allows it to carry oxygen efficiently within the blood
• Describe the structure of hemoglobin with respect to its ability to
bind oxygen
• Describe the basic structure of a white blood cell and explain how
this structure allows it to defend the body against pathogens
• List the 5 major types of white blood cells and classify them as
granular or agranular
• Describe the basic structure of a platelet and explain how this
structure allows it to aid in blood clotting
• List the general location where blood cells are produced within the
body
• List the organs responsible for the recycling of old or damaged
blood cells
• List the 8 major types of ABO/Rh blood types and state the
antigens and antibodies present
• Predict whether agglutination will occur during a blood transfusion
given the ABO/Rh blood types of a blood donor and recipient
• List the ABO/Rh blood type that is the universal donor and
universal recipient
• TIP: The weekly quiz, practice sheets, and other online activities
can give you feedback on your knowledge of this topic and can
help you to feel confident that you are prepared for the lab.
Agenda:
• Upon entry:
• Students will be inspected for proper PPE and lab attire
• Safety considerations specific to this lab will be announced
• This blood used in this lab is simulated and not real.
• During the lab:
• Blood Building – Students will simulate how to build blood by combining
various ingredients together to present blood plasma and the cells
within.
• Blood typing – Students will be given various samples of blood including
a positive control (ABRh+), a negative control (O-) and samples with
unknown blood types. Guided by their instructor, students will use anti-
A antibodies, anti-B antibodies and anti-Rh antibodies to confirm the
blood types of the controls and deduce the blood type of the unknown
samples. List of Terms includes: antibody (anti-A, anti-B and anti-Rh),
antigen (A, B, Rh), blood type (Type A, Type B, Type AB, Type O),
agglutination, red blood cell, blood plasma.
• Appendix: Procedure – Determining Blood Types
• Blood Compatibility - The instructor will demonstrate a method for
determining if blood is compatible for blood donation. Then students will
use their data from the blood typing section to determine if one patient
blood can be successful given to another.
Additional Policies
• Taking pictures
• Students are invited to take pictures of their learning for future study
• Safety
• Students are expected to follow all safety procedures as outlined in the
Lab Safety Module and your instructor
APPENDIX: Building Blood
Make blood plasma:
1. Add the following to an Eppendorf tube:
Ingredient Blood Component Amount Equipment
The biggest component of blood
DI water 500 uL P1000
is water.
Table This symbolizes the “nutrients”
0.5 g Balance
sugar (E.g. glucose)
This symbolizes the “ions” (E.g.
Salt 0.5 g Balance
NaCl)
Protein This symbolizes 3 major proteins
0.5 g Balance
powder – albumin, globulin, fibrinogen.
2. Close the cap on the Eppendorf tube, and shake rigorously for 1 minute. This is meant
to both dissolve the sugar/salt/protein, but also to add gas bubbles from the air such as
oxygen and carbon dioxide.
Add the formed elements (living cells) to the blood plasma
3. Add the following to the Eppendorf tube:
Ingredient Blood Component Amount Equipment
This symbolizes the “red blood
cells.” Note that they are the
Sprinkles 10
most abundant of the 3 types of None
– red sprinkles
cells. This is also what gives
blood its red colour.
This symbolizes the “white blood
Sprinkles - cells.” Note that white blood cells 1
Balance
white are quite rare in comparison to sprinkle
the red blood cells.
This symbolizes the “platelets.”
Poppy Tiny
Note that they are the smallest of Balance
seeds pinch
the 3 types of cells.
Centrifuge the blood
4. With the help of your instructor, add your blood to the centrifuge and spin your sample
for 30 sec at 500 G. Inspect your sample afterwards to see if the plasma and formed
elements have been separated and which of the 2 is the most dense.
APPENDIX: Procedure – Determining Blood Types
Introduction
This protocol describes how to determine the ABO and Rh blood type if given an
unknown sample of blood. Health-care professionals use techniques such as these to
ensure safety when blood transfusions are required.
Procedure
1. Before the blood type of an unknown sample can be determined, it is important to
review the following:
a. Antigens on the red blood cell determine the blood type
Human blood can be Type A, Type B, Type AB or Type O as well as Rh
positive or Rh negative. These names are given based on the presence of
specific antigens on the surface of the red blood cells. See the cartoon
image below to see what the antigens look like for all 8 types:
b. It is possible for blood to undergo agglutination
Healthy blood is smooth, i.e. the red blood cells are separated from each
other within the blood plasma. It is possible, however, for red blood cells to
clump together in in a process called agglutination, resulting in a life-
threatening condition. Below are images of healthy blood that has not
agglutinated and unhealthy blood that has agglutinated. Notice that
agglutinated blood appears “curdled” or “grainy.”
c. Agglutination occurs when certain antibodies are present.
Agglutination only occurs if specific antibodies are present that can bind to
the antigens on the surface of the red blood cell. Importantly, only certain
combinations of antigens and antibodies result in agglutination. Read the
rules below to see how this works:
RULE 1: Agglutination occurs when antibodies bind to antigens
on the surface of a red blood cell.
RULE 2: Blood undergoes agglutination in these 3 scenarios:
• Anti-A antibodies bind to A antigens
• Anti-B antibodies bind to B antigens
• Anti-Rh antibodies bind to Rh antigens
2. Luckily, the principles of antigens, antibodies and agglutination can be harnessed to
determine the blood type of an unknown sample in the lab. This is because blood
from a patient can be combined with lab-made anti-A antibodies, anti-B antibodies
or anti-Rh antibodies and assessed to see if agglutination has occurred. This
analysis can be used to determine the blood type.
See below for 2 examples:
Patient 1:
Patient 2:
3. Now that you understand the theory of blood typing, you can perform it yourself
in the lab on 2 unknown samples of blood. To do this, follow the protocol
below:
NOTE: All unknown samples are tested alongside a positive control (ABRh+)
and a negative control (ORh-).
a. Obtain 4 different 3-welled plates and label them as follows:
i. Plate 1 = Positive control blood (ABRh+)
ii. Plate 2 = Negative control blood (ORh-)
iii. Plate 3 = Unknown patient blood 1
iv. Plate 4 = Unknown patient blood 2
b. Add the antibodies to the 3-welled plates using a transfer pipet as follows:
i. Add 1 drop of Anti-A antibody to the A well
ii. Add 1 drop of Anti-B antibody to Well B
iii. Add 1 drop of Anti-Rh antibody to Well Rh
c. Add the patient blood to the 3-welled plates using a transfer pipet as follows:
i. Add 1 drop of positive control blood (ABRh+) to all 3 wells of Plate 1
ii. Add 1 drop of negative control blood (ORh-) to all 3 wells of Plate 2
iii. Add 1 drop of unknown patient blood 1 to all 3 wells of Plate 3
iv. Add 1 drop of unknown patient blood 1 to all 3 wells of Plate 4
d. Mix using separate toothpicks and wait 10 minutes.
e. Observe if agglutination (blood clumping) occurred in each of the wells.
Record your results in the Table provided.
f. Use your data to determine the blood types of the 2 unknown patient
samples. NOTE: If you are unsure if agglutination occurred, compare your
samples to the positive and negative controls. Remember that the positive
control should have all 3 wells agglutinated and the negative control should
have 0 wells agglutinated.
g. Wash all transfer pipets and 3-welled plates as necessary
