t

) Cell Biology Laboratory Manual with Cancer Applications :,,',

li;r,,'
:,-,.,
ill S
,
,
)
)
; 7. Laboratory Exercise
,
t Blood cells: understanding the basics. Red blood cells,
t white blood cells, and platelets. How they appear and
t how to distinguish them.
)
) lntroduction
t Students are often confused when they learn that blood is a connective tissue. The
t word tissue sounds like a solid component and yet the liquid blood is called a tissue.
t Although blood is a liquid connective tissue, it has many solid components called
t formed elements that are suspended in the liquid part of the blood, called plasma.
) Plasma consists mainly of water with many dissolved solutes. A few substances
, that are dissolved in the plasma include proteins like albumin, globulin, fibrinogen,
, and other regulatory substances like hormones and enzymes. Nonprotein nitrogen
t substances include urea, uric acid, and creatine. Nutrients include amino acids,
) glucose, fatty acids, and glycerides. ln addition, blood contains dissolved gases,
) such as oxygen, carbon dioxide, and various electrolytes, such as sodium,
, potassium, magnesium, and calcium.
,
,
)
t
r,
ffitr ffi
.l:{ffi

r, 'i#x
ffi
t
trr
t ffi;ffi#.P q#u$#-g&H#ffi,
tr, mffi x#ffiffiffiffiffi
.$.&

-i9.7-1. Human blood smear at low-power magnification showing erythrocytes

rt (x 200).

I'
It
!t
It
r' 95

t
i7$ Cell Physiology Lab

Fig.7-2. High powermagnification of a blood smearshowing donutshaped red blood

cells with no nucleus. ln the center is a nucleated white blood cell (lymphocyte) (X
1000).

The solid part of the blood (formed elements) consists of the following:
. Leukocytes (white blood cellsMBC): These cells are of various sizes with
a distinct nucleus (see Figures 7-2 and 7-3).
. ThromboQyteq (platelets or cell fragments): These elements are scattered
within the red blood smear and appear as irregular specks or particles (see
Figures 7-1,7-2, and 7-9).

The leukocytes, depending on their appearance, are classified into two types:

1 . Granular leukocytes (granulocytes).

. Neutrophils (Figs. 7-4 andT-5).
. Eosinophils (Fig. 7-6).
. Basophils (Fig. 7-7).
I

Cell Biology Laboratory Manual with Cancer Applications

r itiff,

2. Agranular leukocytes (agranulocytes).

. Lymphocytes (Fig. 7-B).
. Monocytes (Figs. 7-9 and 7-10).

+,+ffi$t$$f-
, ':1 ,n.t:..,:, , r;;:;:::, r,'
:t!!ij!,i"': .'::1,'::'
,,..,',.,,f11 . ,r :.,,::::iii. '
,,-.,,,,,
f'.11,::::,:i.
.
t:, t.i'':
.;r,, ;. iiit:*$
*.,, ,
'i1
';;, i-*****i
liiiitiiiii,l,ffiffii$$ii+iiifii:.fxl r+r.rni*fi+gf61iry
#ffi$$f fit$f s*.l]:
#++$#s
fiEH$Efi $$x#$i

ff*
ffiffi*--* *$-',fi
ffi H-.
Fig. 7-3. Low magnification of a human blood smear (X100). Note the minute
red blood cells spread like a lawn. The deeply stained cells are white blood cells
(leukocytes)scattered among the red blood cells.

Fig.7-4 illustrates a human blood smear showing a lawn of red blood cells at
high power. This normal smear preparation is made from the author's (LA) blood
with a clear neutrophil in the center.

97
 f
I
78,';;,,,,.,,...! Cell Physiology Lab
I
I
1

I
I
t
I
I
t
I
I
I
I
I
I
I
Fig.l-4. A lawn of RBCs with one neutrophil in the center of the field indicated by
the arrow (X 400).

Fig. 7-5. A neutrophil polymorphonuclear cell. Notice the nuclear lobes attached
by thin nuclear threads (X 2000).

98

I
t
II Cell Biology Laboratory Manual with Cancer Applications i' : ..: 7O:

It
i
t
r,
l,
t
rl
It
r,
t
I.
Ir
Fig. 7-6. An eosinophil with a dumbbell shaped nucleus with a clear connecting
nuclear thread (arrow). Note the granular nature of the cytoplasm (X 1000).

---*-ffiffiffiffi
..:: :'. "':"""::::::" .Ll:,..: ..: ::::: ::::'

Fig.7-7 . A deeply stained granular basophil (X 2000).

99
 r::ii!.,1 Cell Physiology Lab
.,3

Fig. 7-8. A lymphocyte (X 2000).

flg 7-9 A monocyte (X 1000). Note also the small cell fragments or platelets
(arrows) scattered amongst the red blood celts.

100
 Cell Biology Laboratory Manual with Cancer Applications
8.

Fig.7-10. A monocyte, rymphocyte, and a donut-shaped erythrocyte

(x 2000).

What is hematopoiesis?
The process by which the formed elements differentiate into distinct
cell types
is called hemopoiesis or hematopoiesis. Hematorogy is the study
of brood. The
blood constitutes one type of body fluid. Bodily fluids are extremety
important and
play major roles in the body's homeostasis. Blood is important
for homeostasis,
transport of nutrients and gases, and for the body's defense.

How are blood cells synthesized in the body?

All of the different cells in blood and lymph are derived from
a common precursor
cell called the pluripotent stem cel/. These cells are found in
the bone marrow.
Approximately 0.05 to 0.1% of bone marrow cells represent
hemopoietic stem
cells' These cells replenish themselves and give rise to all the formed
elements
of the blood. The pluripotential stem cell generates two distinct intermediary
stem cells, called multipotential stem cells, which include myeloid
stem cells and
lymphoid stem cells. The myeloid stem cell gives rise to
erythrocytes, granular
leukocytes, monocytes, and thrombocytes. The lymphoid
stem cell, on the other
hand, gives rise to T-lymphocytes and B_lymphocytes.

l0t
F
li..i ii ::ti,',:.,ii:,i,i:ii,
; ti: tt:,: ii,i;i ir:r: rt:,f:i T
itt.',! Cell Physiology Lab T
I
f
I
T
I
Multipotential stem cells form progenitor cells, which are also called colony I
forming units (CFU), and each CFU gives rise to distinct, specific formed
elements I
in the blood. For example, erythrocytes have their own colony forming
cells called t
CFU-E. Be aware that stem cells and progenitor cells cannot be distinguished I
morphologically under the microscope, and they often appear like lymphocytes. T
The progenitor cells develop into precursor cells, known as blasfs. you wilr I
notice a number of blast cells in circulation, which will develop into actual
formed T
elements after several rounds of cell division. T
The basics of hematopoesis (how blood cells differentiate from stem cells)
t
are I
shown in a simple flow chart diagram (Fig. 7-11).
I
U
U
Pluripotent cell I
n t
{}
Multipotential cell
I
U
Myeloid series lymphoid series
U
n
il il I
Lt ttilw I
Colony forming Units CFU Precursors of B and T cells U
JI il
{) !
Blast cells
o
B and T lymphoblast
{!
I
RBCs, platelets, monocytes,
B lymphocyte T lymphocyte U
neutrophils, eosinophils, basophils
I
Fig.7-11. Flow chart of hemopoiesis and differentiation of blood cells.
I
I
U
f
U
I
T
I
r
r
102
lr
a
 L.),:a:1::.a::: ::)a:aa':a::ai : a:a:a1 a:ita::ta.t! tai

i: : i : :,: la : l:rii:r: ::::r:iiir:riiti!::irii I

Cell Biology Laboratory Manual with Cancer Applications it rttili8*iil$i+

1 ---^--^'--^*
i

Blood cell cancer

As shown in the illustrations, many blood cell types are suspended in the liquid
part of the blood. Any of these cell types can become abnormal, lose their growth
control, and become cancerous. Depending on the type of blood cells and the
pathological change, they are given different names.

Leukemia's and lymphomas are cancers of the blood and lymphatic systems.
These two cancers together account for an estimated B% (82,000 cases annually)
of cancer incidence in the United States. They are the most common cancers in
children, representing about half of all childhood malignancies (Cooper 1993).

Leukemias and lymphomas result from the continuous proliferation of cells that are
blocked at various stages of differentiation into specialized cell types. Leukemias
arise in the blood forming cells of the bone marrow and can result from abnormal
cell proliferation of any of the different kinds of blood cells within either the myeloid
or lymphoid lineages. Lymphomas, by contrast, develop from lymphocytes or
macrophages in lymphatic tissues (Cooper, 1993).

ln children, leukemia is about twice as frequent as lymphomas, whereas lymphomas

are more common in adults. You will study the various blood cancers both in adults
and children in later sections of this manual. You will also study how leukemia and
lymphoma are classifled, the blood cell types involved, and how the pathology of
the bone marrow changes in disease in the histopathology section of this manual.

How do you identify various types of blood cells?

Microscopic analysis of blood cells and evaluation of blood cell
pathology.

When students are given microscopic glass histology slides of human blood
smears, they often complain of the following technical problems:

1. They see nothing on the slide and believe it is a blank slide.

103
 84 i) .......
Cell Physiology
_.
Lab

2. They claim the microscope is not working since nothing comes into focus.
3. They claim they cannot see anything because it is too small.

Fig.7-12 shows a low-power magnification of a human blood smear. Blood cells

appear like little specks under low power. Students often fail to focus on these
cells because they rotate the course knob of the microscope too quickly.

Fig.7-12- A low power magnification of a brood ceil smear (x 40).

f- The illustrations above were presented to familiarize students with various blood
cell types before moving to the next section, which presents a macro blood cell
visual model. This model is designed to better help you understand blood cell
anatomy and what can go wrong in disease states. ln addition, the macro model will
help you better visualize microscopic materials and to understand the hemopoietic
system and defects in blood cells leading to various changes in histopathology.
For more details, see leukemia and lymphoma under the histopathology section
of this manual.

t04
 cell slolosll t-aboStoy Malual with cancelAnnlicatiols
.s

In this laboratory exercise, students should be able to identify the blood cell types
by using vendor-supplied blood histology slides.

Laboratory Exercise:

Observe the following blood cell types under the microscope, and draw them in
color in the space provided. See illustrations for guidance. Start with low-power
field, and then move to high-power magnifications. Your instructor will guide you in
the proper handling of the slides and identification of cell types.

Erythrocytes

Thrombocytes

Neutrophils

Eosinophils

Basophils

Lymphocytes

Monocytes

Blast cells

105
 Cell Physiology Lab
i+#,;

Draw various tyoes of blood cells below and verify with your instructor

Red blood cells Platelets

Neutrophil Eosinophil

Basophil Lymphocyte

Monocyte A blast cell

6ta
Did you know that you can make your own blood slide from a pin drop of blood?; il
[See procedure at the end of section ZB.]

106
 Cell Biology Laboratory Manual with Cancer Applications
liiiirS#;

Critical thinking:

't. were you able to identify blood cell types under low power magnification?

2. How many types of white blood cells are there?

3. What are the functions of white blood cells?

4. can something go wrong with white blood ceils? (Hint: what is leukemia? How
many types of leukemia are there? List alltypes of teukemia that you are familiar
with, and identify the bload cell type involved in each teukemia.)

5. ln leukemia, do blood cells increase or decrease in number? Do blood cells

change in their shape?
Cell Physiology Lab

tn
,G-
T
) i Something more to think about:

6. Have you ever wondered why there is a critical shortage of blood for transfusion?

7. Who can give blood as a donor? What qualifies a person? (

I
I
I
I
{
B. What is a blood transfusion and how is artificial (synthetic) blood made?
(
I
I
(

i08