8

LABORATORY ACTIVITY II
CYTOLOGY

A. ANIMAL CELLS
The cell is the basic unit of all organisms. Most cells are microscoplic, though some
Iike bird's eggs are visible through our naked eyes, Although all cells are basically the same,
they differ in detals of thelr structures. A typlcal anlmal cell Is irtually non-exdstent. These
diíferences are correlated with the specific task of the cell as a part of the organism. The
visible structures of the cell
are called organelles.
Scrape the inside of your cheek with the blunt end of the toothpick. Place the
Scraplngs on a slde with a drop of lodine solution and cover with a glass slip. Examine the
CHEEK CELLS (Figure 2.1a) under the LPO and HPO. If the cells have absorbed the iodine,
the nucleus will appear as a small round dark body near the center of each cell. The
cytoplasm is lighter and bounded by the plasma (cell) membrane.
The FROG'S ovUM (FIgure 2.1b) is surrounded by a cll (plasma) membrane which
encloses all other organelles. The nucleus is usualy round and bounded by a smooth or
comugated nuclear membrane contalining several nucleoli along its periphery. This membrane
encloses the nucleoplasm. Between the plasma membrane and the nuclear membrane is an
area flled with cytoplasm.
ULTRASTRUCTURES OF A CELL (Figure 2.1c) Because of its extraordinary
resolving power, the electron microscope seems to be the ideal instrument for the study of
cellular ultrastructures. Direct magnifications as high as 20,000x to 160,000x may be obtained
and the micrographs may be enlarged photographically to 1,000,000x or more depending
upon the resolution achieved. The following structures are commonly revealed in detail under
the electron microscope:

1. The PLASMA (CELL) MEMBRANE is a thin film surrounding the cell. It is

actually a double-structured membrane having lipoidal and protein
components which is about 100A° thick. The membrane is capable of
lowering the rate of penetration of molecules as well as discriminates
between them having the property of selectively semipermeable. A number
of modifications of the cell membrane is apparent. The narrow evaginations
or finger-like projections distributed at random along the cell surfaces and
with the oytoplasm are called microvilli. In the striated border of the
intestinal epithelium the microvilli are packed to form a continuous surface.
Some cells are diliated. The ilia terminate in a transverse plate called basal
body. Individual dilium is made up of a ring of nine peripheral and two
centrally located filaments running in the direction of the long axis. The
narrow invaginations of the cell surface are called pinocytic vesicle for the
ingestion of dissolve substances. Localized thickenings of apposed cell
membranes have been observed in many cells. The thickened portions of
each membrane are generally very close together appearing as a single
structure called desmosomes. The basal membrane is commonly formed by
connective tissues and amorphous ground substance.

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2. The CYTOPLASM flls the space between the cell membrane and nucleus. It
s primarly protelnaceous and many of its properties similar to the colloidal
dlsperslons of complex protelns. The colloidal properties essental to sol-gel
transformatlons, viscoslty, splndle formatlon, and cell cleavage.
3. MITOCHONDRION IS a double-membraned structure where the outer
membrane is stretched tighty around while the inner membrane
Invaglnates Iinto the body of the organelle forming serles of compartments
or cristae. Whether the cristae are invaginations or separate structures are
still debatable. This is the center of cellular respiration where the foods are
oxldized to CO2 and H0 with the subsequent release of energy. The
complete Kreb's Cycle is probably carried out in the mítochondria.

4. The LYSOSOMES are spherical bodles containing a number of digestive

enzymes which break down biological polymers into smaller constituents
that can be processed by the mitochondrial enzymes. The lysosomal
membrane Isolates the digestive enzymes from the rest of the cytoplasm.
Ruptures of the membranes and consequent release of the enzymes lead to
the lysis of the cell.

5. The CENTRIOLES are cylinders which are open at both ends. The wall of
centriole has nine groups of microtubules arranged in a circle. The central
microtubules and special arms are absent in the centriole.

6. A system of flattened sacs which may dilate to become vacuoles is the

GOLGI COMPLEX. One of the main characteristics of the complex is the lack
of ribosomes. The Golgi complex appears to be surrounded by a zone from
which the ribosomes are excluded. Protein synthesls does not occur in this
organelle. The structure is concerned with the storage and alteration of
substances synthesized in the cell and their transport in the Golgi zone.

7. The ENDOPLASMIC RETICULUM are channels of vesicular bodies

interconnected to form a complex network which separates two distinct
phases In the cytoplasm. The rough endoplasmic reticulum is well
developed in cells whlch are actually engaged in protein synthesis. RNA
containing ibosomes are attached to their membranes. The smooth
endoplasmic reticulum forms a continuous system making up a tubular
network that pervades the cytoplasmic matrix.

8. The NUCLEUS may be fusiform, ellipsoidal, flattened or irregular depending

on cell shape and function. The following structures are recognized in the
Interphasic nudeus:

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(a) A nucdear membrane appears as a clear outine on both ytoplasmic

and nuclear sldes. This Is composed of two membranes showing
openlngs or nucleer pores.
(6) The nucleoplasm flls most of the nucleer spece. This represents the
uncondensed reglons where the chromosomes are largely dispersed
into fine macromolecular components.
(c) The condensed regions of the chromatin representing the parts of the
chromosomes are frequently found near the nuclear membrane and
partly attached to the nucleolus.
(d) The nucleolus is generally spheroidal and large. They are single or
multiple
usualy containing ribo-nucleoproteins.
Label all the visble parts of the cheek and egg cells. Label the parts of an ideal animal cell as
seen under the electron microscope.

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Name: Date:
Section: Room
LABORATORY ACTIVITY 2A
CYTOLOGY Animal Cells

Figure 2.1a. Cheek cells from the human mouth. Figure 2.1b. Egg cell of the
frog.

Figure 2.1c. General diagram of the ultrastructure of an animal cell.

Questions:
1. What are organelles? Enumerate all organelles seen in your specimens?

2. Does a typical cell exist? Why?

3. How does an animal cell differ from a plant cell1?

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B. SHAPES OF CELLS

The variety of shapes assumed by cells is largely determined by their arrangement,

position, and function within their respective tissues or organs. There are different shapes of
cells and some of them are as follows:
a. The egg cells or ova of frogs are rounded with big centraly located nuclel gving
the shape of SPHERICAL. The nucleoli are numerous, and the cytopasm occupies
the space between the nucleus and cell membrane.
b. The STELLATE cells are motor cells of the dorsal and ventral horns of the frogs
spinal cord. They may be star-shaped or triangular in appearance with one to
several protoplasmic processes and are essential for the conduction of impulses
from the spinal cord to different parts of the body.
C. The flattened, hexagonal cells is called SQUAMOUS which is located at the outer
layer of the skin for protection, they are closely set together Ilike the tles of
mosaic pavement.
d. The cells occur in the epithelial tissue along the inner lining of the small intestine
is called COUMNAR. The height exceeds the width and the nucleus is distincty
oval. They are responsible for the absorption of nutrients.
e. The PYRAMIDAL cells are found at the tip of the loops of the inner lining of the
small intestine; its shape is similar to an isosceles triangle. Its function is the
same as that of the columnar cell.
f. The cels that constitute the muscular layer of the intestine or stomach are the
FUSIFORM ells. They are spindle-shaped, tapering at both ends, and responsible
for peristalsis.
g. Equidimensional cube cells that line the uriniferous tubules of the kdney are
CUBOIDAL in shape. The lumen formed serves as the passageway of urea as
fitered by the Bowman's capsule.
h. Liver cells are distinctly POLYGONAL. They are responsible for the secretion of
blle.
i. The shapes of white blood corpuscles of the frog are variable and imegular which
is said to be AMORPHOUS due to their activities. They enguf bacteria and other
forelgn bodies.
. The red blood cells are clearly OVAL wlth prominent nuclel. They carry
hemoglobin and oxygen to the tissue from the lungs.

C. OSMOSIS IN RED BLOOD CELLS

Living cells are dynamic units which they rely on chemical and physical
processes to carry out many functions. Every functioning cell participates in the
maintenance of a dynamic equilibrium or homeostasis within the entire organism.

Prepare three (3) slides containing fresh blood smear of the frog. On one
slide place a drop of distled water (hypotonic solution) and on another a dnrop of
physlological salt solution (0.9 % NaC) which Is isotonic. On the third slide put a drop

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or so of 3% NaC (hypertonic solution) along one edge of the cover slip. Observe the
shapes of the cells after a few seconds.

Label the parts of the different klnds of cells. Accomplish the table of the source and functions
of the different types of cells.

Draw the red blood cells as affected by the three kinds of solution.

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Name: Date:
Sectlon: Room:

LABORATORY ACTIVITY 2B
CYTOLOGY Shapes of Cells

Fig. 2.2a. Sphercal cell Fig 2.2b. Stellate cell

Fig. 2.2c. Squamous cell Fig. 2.2d. olumnar cell

Fig. 2.2e. Pyramidal ce Fig. 2.2f. Fusiform cell

Fig 2.2g. Cuboidal cell Fig. 2.2h. Poygonal cell

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Fig. 2.21. Amorphous cell Fig. 2.2e. Oval cell

Sources and Functlons of Varlous Types of Cells

Shapes of Cells Sources Functions

1.Spherical

2.Stellate

3.Squamous

4.Columnar

S.Pyramidal

6.Fusiform

7.Cuboidal

8.Polygonal
Amorphous

10.0val

Questions:

1. What are the criteria for classifying the cells?

2. Are all cells microscopic? Defend your answer.

Laboratory Guide in General Zoology-

Name: Date:
Section: Room:
LABORATORY ACTIVITY 2C
CYTOLOGY Osmosis in Red Blood Cells
Appearance and Reacton of Red Blood Cells in:

Appearance Reaction
1.Hypotonkc Solution

2.Isotonic Solution

3.Hypertonic Solution

Questions:
1. What is the importance of diffusion in cells?

2. Why do cells behave normally in 0.9% salt solution?

3. Explain the reactions of red blood cells to the solutions.

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D. MITOSIS

Cell division can be classified as direct or indirect. In mitosis (indirect), a

spindle is formed and the chromatin of the nucleus Is organized into chromosomes
which split
longitudinally
unlike in amitosls (direct) where the nucleuS simply
constricts into two parts. Mitosils is the usual type of cell dvislon while amitosis is
relatively rare. From algae to orchids and amoeba to man the essental processes of
cell dviskon are remarkably simllar. We can demonstrate the divislon of one cell and
illustrate the fundamental strategy In al.
The cells of the whltefish (Coregonus) blastula are used to ilustrate cdearty
the process and reveal the differences distingulshing animal from plant mitosls. The
process In both cases is basically simlar, except that in animal cels, centrioles with
radlating astral rays are present and the partition between daughter cells is produced
by a constriction or furmrOw. On plants, the centrioles are absent and the partitions
bebween the cells or plates are formed between the daughter nuclel. However, the
behavior of the chromoscomes is fundamentally the same. The chromosome number
of various species of Coregonus is gven below. The chromosomes are numerous,
darkly stained, and their behavior can be studied easily by beginning students.
Examine sections of whitefish blastula and identify the following stages.

Table 1. Chromosome Number of Various Species of Coregonus (Zelinsky & Makhrov,

2002).
Species Chromosome numbers
2n
Coregonus albula 80
Coregonus lavaretus 80
Coregonus lavaretus baerd 80
Coregonus schinzll 72 36
a. INTERPHASE (Fig. 2.3a). Ths Is the normal cell between successive mitosis.
This is the perlod of exposure of genetlc materials, proliferatlon of RNA and
proteins in the cytoplasm. The term "resting cell" is not accepted because the
cells are very active, manufacturlng protoplasm for cell growth and synthesizing
proteins for the next cell division. The nucleus is enclosed by a nuclear membrane
and contalins chromatin which forms an Iregularly scattered granular network
with one or more nucleoli. The centriole with radiating astral rays are clear.

Fig. 2.3a. Interphase

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b. In PROPHASE (Fig. 2.3b) stage, the chromosomes become distinct as long thin
threads which gradually thicken as they split longitudinally into two halves which
IS called chromatids. Somewhere along their length the chromatids are joined by
a spherlcal, refractle body called centromere (kinetochore, chromomere,
chromocenter), Chromatids continue to shorten coils decrease in number, as they
are Increasing In diameter. Simultaneously, the centriole divides Into two and the
halves migrate along the nuclear membrane until they lie opposite each. The
moving centrioles, as they travel toward the poles, acquire a number of radiating
refractile fibers in a starburst pattern called asters, and the fibers are astral rays.
Toward the end of this stage, the nuclear membrane and the nuceus
disintegrate.

Fig. 2.3b. Prophase (Ear, Middle, Late/Prometaphase)

C. The nuclear membrane has disappeared and the chromosomes are grouped along
the equatorial plane of the cell midway between the poles during the METAPHASE
(Fig. 2.3c) stage. Spindle flbers run betweern the centrioles. This is the shortest
phase of mitosis. This stage is also marked by dissolution of the nuclear
membrane and the appearance of astral rays.

Fig. 2.3c. Metaphase

d. Immediately after metaphase in which the chromosomes lie along the equatorial
plane, the centromeres split and the daughter chromatids begin to migrate
toward opposite poles which indicate the beginning of ANAPHASE (Fig. 2.3d). The
movement of the chromatids is accomplished by two simultaneous and
complementary actions. The spindle fiber connecting the centromere with the
centriole is a contractile protein and its contraction pulls the chromatid. Another
fiber develops between the halves of the centromere and pushes the chromatid.
Movement of chromatids may be due also to mutual repulsion of the division

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Products of each centromere. The result is a duster of chromatids assembled

around each centriole..

Fig. 2.3d. Anaphase

e. In TELOPHASE (Fig. 2.3e), the chromatids reach their polar destination. Each
chromatid becomes less distinct and the spindle fades. The nudear membrane is
reconstituted and the nucdeus reappears. This stage s accompanled by a divislon
of the cytoplasm (ytokinesis) as shown by the appearance of a deavage furrow
at the equatorlal plane of the cell. The mechanism of cytokinesis is still unknown
but evidences point that asters and astral rays play a vital role.

Fig. 2.3e. Telophase

f. TWO-DAUGHTER CELL (Fig. 2.3). The cleavage furrow deepens until complete
cytoplasmic separation is achieved. Each daughter cell s actually an interphasic
cell.

Fg. 2.3f. Two-daughter cll

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MORPHOLOGICAL CHANGES DURING ANIMAL MITOSIS.

changes oCcurring in varlous cellular organelles can be convenienty summarizedmorphologca
Some -

as TOowS:

Structures Stages of Mitosls

1.Nuclear InterphaseProphase
Intact
Metaphase_ Anaphase Telophase
Disintegrating Disintegrated Disintegrated | Reforming9
Membrane
2.Necleoll Visible and Dispersed Invislble Invislble Invisible
darkly
stained
3.Chromatdds/ | Randomly Condensed Allgned along | Split at Aggregatng
Chromosomes colled equatorlal centromeres, at poles,
threads plate migrating indistinct
toward poles Invisible
4.Centromeres Invisible Visible as dark | Visible as Split Invisible
dots dark dots
5.Centrioles Visible Form Remain at Remain at Start
spindles, poles poles replication
migrate
towards the
poles
6.Spindle Not formed Being formed Attached to Visible Disappear
Fibers centromeres between
centrioles
Compare the mitobic stagesof whitefish blastula with the photomicrographs (Figs. 2.3 a-f).
Label the different parts of the cell as It undergoes mitos!s.

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Name: Date:
Sectlon: Room:.

LABORATORY ACTIVITY 2D
CYTOLOGY Mitosis

Fig. 2.3a. Interphase Fig 2.3b. Prophase

Flg. 2.3c. Metaphase Fig. 2.3d. Anaphase

Fig. 2.3e. Telophase Fig. 2.3f. Two-daughter Cell

Questions:
1. What causes the cell to divide?

2. Briefily differentiate the varlous stages of animal mitosis from each other using the
behaviour of the chromosome as the basis,

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