1. Human Anatomy and Physiology
Lab 1
Blood

2. Background:
I. Blood is a connective tissue composed of formed elements (cells and cell fragments) and intercellular material (plasma)

3. Blood Sample Centrifuged
adult males: 5-6 liters
adult females: 4-5 liters

4. Hematocrit:
the portion (% by volume) of the blood that is composed of packed red blood cells
adult males: % + 5%
adult females: 42% + 5%

5. Objective 1 Plasma Characteristics
Plasma is the extracellular (intercellular) material of the connective tissue, blood:
Characteristics:
90% H2O
10% solutes:
plasma proteins (albumin, antibodies, clotting proteins)
nutrients (eg, proteins, carbohydrates, lipids, vitamins)
hormones
wastes (eg, urea, uric acid, creatinine)
dissolved gases (CO2, O2)

6. Lab Objective 1:
In this exercise, you will obtain a sample of sheep plasma and determine its:
color
transparency pH
consistency

7. Formed Elements: cells and cell fragments
Leukocyte
Erythrocyte
Platelets
Erythrocyte/Red Blood Cells (RBCs): anucleate cells that carry oxygen & carbon dioxide X 106 cells/mm3 blood
Leukocyte/White Blood Cells (WBCs): nucleated cells that provide immunity ,800 cells/mm3 blood
Platelets (thrombocyte): cell fragments used for hemostasis ,000 – 400,000 cells/mm3 blood

8. Red Blood Cells (erythrocytes):
- lack nuclei and organelles
Contain hemoglobin (Hb) which binds to O2
(oxyhemoglobin ) & CO2 (carbaminoglobin)
- have a “biconcave” shape
- diameter averages 7.5 m.

9. Never Let Monkeys Eat Bananas
There are five types of leukocytes organized into two classes:
Never Let Monkeys Eat Bananas

10. Platelets (Thrombocytes):
- fragments derived from bone marrow cells (megakaryocytes)
contain granules but no nuclei or organelles
prevent blood loss (hemostasis)

11. Objective 2
You will identify each of the formed elements on a prepared blood smear that has been stained with Wright’s stain
Wright’s Stain is a mixture of two dyes:
1. Methylene Blue: a basic dye that stains acidic components deep blue/purple
2. Eosin: an acidic dye that stains basic components red/deep pink/orange

12. Methylene blue (blue dye)
Different stains can be applied – one popular stain is a differential stain called Wright’s Stain. Wright’s stain is a mixture of eosin and methylene blue.
Methylene blue (blue dye)
has a positive charge and stains negatively charged substances (acids found in some granules, and DNA and RNA) structures that combine with methylene blue are called basophilic
Eosin (red dye)
has a negative charge and stains positively charged substances (bases found in some granules, and hemoglobin) structures that combine with eosin blue are called acidophilic
Eosin/Methylene Blue Complex
stains neutral substances (such as components of some granules) lilac

13. When blood is smeared onto a slide, dried and stained with Wright’s stain, the individual formed elements (including types of WBC’s) can be distinguished:

14. Granulocytes: Neutrophil: 50 – 70% of the circulating WBCs
m in diameter
- 2-5 nuclear lobes
- pale staining, lavender granules

15. - 2-4% of the total circulating WBCs
Eosinophils:
% of the total circulating WBCs
m in diameter
- have a bilobed nucleus
- bright reddish/orange/pink cytoplasmic granules

16. - 0 – 1% of the total circulating WBCs
Basophils:
- 0 – 1% of the total circulating WBCs
m in diameter
- unsegmented or bilobed nucleus
- deep blue/purple cytoplasmic granules

17. Agranulocytes: Lymphocytes: - 25 - 45 % of the total WBC count
- size ranges from 5 m (small) to m (large); small lymphocytes predominate
- nucleus is round or slightly oval; it may be indented
- clear blue cytoplasm that may be seen only as a ring around the nucleus
Small lymphocyte
Large lymphocyte

18. Monocyte - 3-8 % of the total WBC count - 14-24 m in diameter
- nucleus is horseshoe shaped or kidney shaped
- abundant blue gray cytoplasm that
may contain vacuoles

19. Platelets appear as small, granular cell fragments(2-3m) that may occur singly or in clumps

20. Objective 3
A differential white blood cell count is performed to determine the relative percentage of each type of WBC
It is used to detect diseases, such as acute infection, chronic infection, allergy, parasitic diseases, anemia, HIV infection, and others

22. In this activity, you will systematically scan a prepared slide and observe a TOTAL of 100 white blood cells
- identify each one and record your data to determine the relative percentages of each type of WBC
# observed
% = X 100
# counted

23. Each of the leukocytes exists within a range of normal values in peripheral blood.
Type of Leukocyte Normal % #/100 Cells If Elevated?
Neutrophil % acute infection
phagocytic
Eosinophil 2-4% allergic reaction, parasitic infection
asthma
Basophil % histamine & heparine
Lymphocyte % Immunity
Monocyte 3-8% chronic infection

24. Activity Blood Typing
Pretested blood samples will be tested for the presence of A, B and Rh agglutinogens
Type A Type B Type AB Type O
Clinically Important Blood Groups Include the ABO System

25. Rh System – which includes the D agglutinogen
Rh+ Rh-

26. Blood Type is determined by which agglutinogens are present on RBC surfaces:
Type A Type AB Type B-
Type AB Type O-

27. Agglutinins are antibodies that combine with agglutinogens and cause a clumping reaction called agglutination +
Type B Cells Anti-B agglutinin
Agglutination

30. To determine the blood type of a sample, mix separate drops of a blood sample with anti A agglutinin, anti B agglutinin and anti-D agglutinin:
Blood Typing Slide With 3 Drops Of Blood To Be Tested, One Drop In Each Well

31. Add Anti-A serum to this well and mix
Add anti-D serum to this well and mix
Make sure that you use a separate toothpick for mixing in each well…………….
Add Anti-B serum to this well and mix
If the agglutinin in the antiserum binds to agglutinogen you will soon observe agglutination

32. For example: Type B+ Type B- Type O+ Anti A Anti D Serum Serum
Anti B Serum
Type B Type B Type O+

34. This is what agglutination will look like !!!!!!!!!!!!!!!!!!!!!!!!

35. Safety Precautions for Dealing With Blood Samples:
Wear gloves
Wear goggles
Do not test your own blood – use our samples
Place any materials that contact blood in bleach solution immediately after use
5. Avoid spilling the samples – do not carry the slides or samples from place to place. If there is a spill, cover with bleach solution immediately and notify your lab instructor

36. Practice Blood Typing