~BASIC CLINICAL SKILL GUIDELINES~

HISTOLOGY
Instructors:

No Name Department Phone

1 dr. I G.N. Sri Wiryawan, M.Repro Histology 08123925104
2 dr. I Wayan Sugiritama, Mkes Histology 08164732743
3 Dr. dr. I A. Ika Wahyuniari, M.Kes Histology 08123614856
4 Dr. dr. Ni Made Linawati, M.Si Histology 081337222567
5 dr. I G A Dewi Ratnayanti, M.Biomed Histology 085104550344
6 dr. I G K Nyoman Arijana, M.Si.Med Histology 085339644145
7 dr. Ni Luh Gede Yoni Komalasari Histology

General Instructions:

1. In each basic clinical skill all students should wear the laboratory cloak!
2. Read the instruction before beginning the basic clinical skill session!
3. It is not allowed to drink or eat in Histology Laboratory
4. Taking picture of the histology slide is permitted.
5. Please handle the microscope and slide with care!
6. Any damage and loss of the microscope or slide will be charge to the student.
7. On having finish observing the slide under the microscope, please draw/print the picture
of each slide and address the important part of the slide as appropriate (with number
and note as the example below)!
8. Each group should collect the completed practical worksheet on the beginning of the
following basic clinical skill schedule.

Instructions on How to Use the Microscope:

1. Turn on the microscope and the light of the microscope.

2. Scroll down the slide desk.
3. Put the slide on the slide desk properly.
4. Use the lowest magnification (4x magnification of objective lens).
5. See through the ocular piece.
6. Scroll the macrometer of the microscope while peeking through the ocular piece until the
tissue is clearly seen.
7. Adjust with the micrometer for better visualization and focus.
8. Adjust the amount of light if needed.
9. Move to the higher magnification step by step, first to 10x objective magnification and if
the tissue is clearly seen than can proceed to next higher magnification.
10. Observation with 100x objective magnification needs an oil immersion to be able to
observe the slide properly.
11. At the end of the basic clinical skill session all slides should be collected to the instructor
and the microscope should be turned off.

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Format for basic clinical skill worksheet :

Cover

Basic Clinical Skill

Histology

UNUD LOGO

SGD….
Member :
Name and Student Number

Department of Histology
Faculty of Medicine Udayana University
2018

Content :

BCS 1. Basic Histology

Slide 1.

Examiner: ………………………………………..

Signature:

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BCS 1 Basic Histology

Epithelium

OBJECTIVES: At the end of this laboratory you should be able to:

1. Recognize and correctly name the types of epithelium.

2. Name one representative site in the body where these types are found.
3. Distinguish between multicellular and unicellular glands.
4. Distinguish between serous and mucous secretory glandular cells.

SLIDES FOR THIS LABORATORY: 1,2,3,4, 5A, 5B, 6, and 7

SURFACING EPITHELIUM

SIMPLE SQUAMOUS EPITHELIUM:

Slide 1 Kidney

Simple squamous epithelium can be studied by looking at cells which line parietal layer
of capsula Bowmans.

SIMPLE CUBOIDAL EPITHELIUM:

Slide 1 Kidney.

Simple cuboidal epithelium is prominent in the cortex and medulla of the

kidney. Proximal convoluted tubules , distal convoluted tubules , and collecting ducts are
examples of simple cuboidal epithelium.

SIMPLE COLUMNAR EPITHELIUM:

Slide 2 Duodenum.

Locate a large fingerlike projection at the surface of the organ (this is an intestinal
villus ). Observe that the projection has an internal component and a surface. The
surface of each villus is covered with simple columnar epithelium . The free surface of
these cells has very tiny projections called microvilli , which are specialized for
absorption (absorptive cells). Numerous mucus-secreting cells, known as goblet
cells because of their shape, are dispersed between the absorptive cells.

PSEUDOSTRATIFIED CILIATED COLUMNAR EPITHELIUM:

Slide 3 Trachea.

Note the ciliated, pseudostratified columnar epithelium lining the tracheal lumen. This
epithelium is commonly referred to as "respiratory epithelium".

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STRATIFIED SQUAMOUS EPITHELIUM (KERATINIZING AND NONKERATINIZING):

Slide 3 Esophagus.

Stratified squamous nonkeratinizing epithelium lines the lumen of the esophagus. This
type of epithelium is found in mucous membranes.

Slide 4 Skin

Observe the thick cornified layer of stratified squamous keratinizing epithelium of the
skin. Note the changes in cellular morphology and intercellular spaces as one moves
from the basal cells to the lifeless keratinized cells on the surface.

Transitional epithelium:

Slide 5 A (Urinary Bladder contacted) dan 5 B (Urinary bladder distended).

Find the transitional epithelium that lines the urinary bladder.

Note the presence of characteristic surface dome cells in the transitional epithelium .

GLANDULAR EPITHELIUM

SEROUS GLANDS:

Slide 6 Parotid.

The parotid gland is a pure serous gland and thus made up of all serous acini . Note the
prominent granules of the acinar cells.

MIXED GLANDS:

Slide 7 Submaxillary gland.

Also organized as acini, the submaxillary gland secretes both serous and mucous
products. Serous cells are more numerous in the submaxillary (also called
submandibular) gland. Mucous secreting cells have flat nuclei located basally. The
apical region typically appears empty due to the extraction of mucous in routine tissue
preparation. Serous cells are often found capping mucous acini. These are
called demulines and are easily distinguished by their intensely staining granules.

.UNICELLULAR GLANDS:

Slide 2 Duodenum.

Observe Goblet cells in the simple columnar epithelium of the duodenum.

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Connective Tissue

OBJECTIVES: At the end of this laboratory you should be able to:

1. Distinguish the connective tissues from all epithelial tissues on the basis of location, cell
density and the presence of discrete fibers.

2. Distinguish between loose irregular (areolar), dense irregular, or dense regular connective
tissues on the basis of fiber packing and orientation.

3. Identify and list the cell types found in the various kinds of general connective tissues, and
describe their origins and functions.

SLIDES FOR THIS LABORATORY: 11, 12, 13,14,15,16,17,18,19,20, dan 21

FIBER OF CONNECTIVE TISSUE

Slide 11 Spleen, Reticular stain.

Reticular fibers (type III collagen) are thin collagen fibers not typically detected with
routine H & E staining. However, these fibers stain black with silver stain and are often
called argyrophilic fibers . The spleen demonstrates the supportive network of reticular
fibers present in many organs. In this slide, the delicate reticular fibers are black and the
thick collagen fibers are red/brown.

Slide 12. Elastic Fiber (Elastic cartilage), Van Gieson stain.

Elastic fibers are slender and branching, are difficult to differentiate with collagen fiber in
standard staining H & E. Special staining to detect elastic fiber is Van Gieson and the
fibers appear black. In this slide you will observe condrocytes in lacunae and the elastic
fibers scattered in between.

Slide 13. Collagen fiber type I (Skin), Picro Sirius Red.

Collagen fiber type I is the most common fiber found in our body, appear pink in routine
H & E. Special staining commonly used to visualize collagen fiber is Picro Sirius Red,
the fiber is stained red. Collagen fibers appear dense and unbranching, Elastic fiber are
not stained and the muscle and hair follicle are yellow.

TYPES OF CONNECTIVE TISSUE

Connective tissue can be classified as either embryionic connective tissue and connective
tissue proper or specialized connective tissue. Connective tissue proper includes: loose
connective tissue (also called areolar) and dense (irregular) connective tissue. Specialized
connective tissue types include: dense regular connective tissue, cartilage, bone, adipose
tissue, blood, and hematopoietic tissue. The majority of specialized connective tissues will be
studied in future laboratories.

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Slide 14 Developing Teeth ( Mesenchymal CT), HE

Developing teeth contains dense mesenchymal cells, which are large and have small
processus. The nuclei are oval and mitotic figures are often seen. Extracellular matrix
mainly compose of ground substances which fill the space between cells, no fiber are
visually detected in this slide.

Slide 15 Umbilical Cord (Mucoid CT), HE

In this slide you can observe umbilical artery and vein. Between the vessels lies the
mucoid connective tissue, which is the transition from embryonic to mature tissue.
Hence you can observe primarily fibroblast cells, with scarce mesencymal cells. The
collagen fibers are sparse and begin to take it’s mature form. Ground substances are
abundant.

Slide 16 Loose CT, HE

In loose connective tissue we can observe many types of cells including fixed cells and
transient cells. In the extracellular matrix the fiber are relatively few than the abundant
ground substance. Mixed types of mature fiber are seen, but mainly are collagen fibers.

Slide 17 Skin (Dense Irregular CT), HE

Dense connective tissue is termed based on the more numerous and closely packed
fiber in the extracellular compartment. The connective tissue in the skin is found in the
dermis. Below the epidermis there is thin loose CT and followed by the thick dense CT.
The cells are primarily fibroblast and scattered throughout the fiber. Commonly all types
of fiber can be found in this type of CT. In this slide we can observe that the fibers run in
many directions.

Slide 18 Tendon (Dense Regular Colagenous CT), HE

Tendon only composed of organized collagen fibers. The fibers are run in uniform
direction which is longitudinal in this slide. The fibroblast cells are lined in the periphery
of the fiber pararel to the fiber direction. The cells are flattened due to the strong
collagen fiber type I.

Slide 19 Ligament Nuchae (Dense Regular Elastic CT), HE

The elastic fibers run in uniform longitudinal direction. The elastic fibers are slender,
branching and elastic hence the fibroblast cells are scattered and maintain it’s original
form with oval nucleus.

Slide 20 Fat (White Adipose Tissue)

The slide mainly composed of fat cells. The unilocular fat cells are round or polygonal.
The nucleus is pierced to the periphery by the the large fat containing vacuole, giving an
image like a signet ring. The framework of adipose tissue is suppored by reticular tissue
which is not visually detected in this slide.

Slide 21 Liver (Reticular CT) HE

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 Liver are compose of liver cell arranged in liver lobule. The framework of the organ is
supported by reticular tissue. The reticular fiber cannot be seen in this slide. The
fibroblast seen in liver are refer to reticulocyte.

BCS 2 MUSCULOSKELETAL

Muscle Tissue

OBJECTIVES: At the end of this laboratory you should be able to:

1. Recognize and describe the anatomical features of muscle types at the tissue, cellular,
and subcellular.
2. Identify and distinguish between the muscle types in cross or longitudinal sections in the
light microscope or in electron micrographs.
3. Identify the subcellular components of myofibers and understand how they contribute to
contractility in each muscle type.
4. Identify and understand the functional significance of intercalated disks in cardiac
muscle.

SLIDES FOR THIS LABORATORY:

Skeletal muscle

Slide …Skeletal muscle, cross and longitudinally sectioned.

Study the striations of skeletal muscle. Locate a site where muscle fibers are
clearly longitudinally sectioned . Compare your observations here with electron
micrographs of skeletal muscle in your text book, particularly the structure of a
longitudinally sectioned sarcomere. Note the location and content of A bands , I bands ,
and Z lines. In the cross-sectioned skeletal muscle of the slide, locate a group of cells (a
fascicle) and look for nuclei near the edge (sarcolemmal region) of the muscle fibers
(cells). Identify endomysium , perimysium , and epimysium .

Smooth muscle

Slide … Duodenum

Find the smooth muscle in the outer perimeter of this section. It is represented here as
two layers, an outer longitudinally oriented layer and an inner circularly arranged layer .
You can readily observe the appearance of smooth muscle cells
in cross and longitudinal profiles .

Cardiac muscle

Slide … Heart

Observes the striations present in cardiac muscle. Also observe fiber bundles which are
cut in longitudinal , oblique and transverse section. Observe the centrally located nuclei
of cardiac muscle. Locate a large area of longitudinally sectioned fibers and note:

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 1. The walls of individual fibers are parallel for short distances only.
2. The variation in width is produced by frequent branching and anastomosing of
adjacent fibers.
3. The branches arising from individual fibers are variable in diameter.

In longitudinal section, the regular branching and anastomosing pattern is the most
distinctive and characteristic feature of cardiac muscle. Higher magnification will be
required to confirm the presence or absence of myofibrils and cross striations .

Observe an appropriate branching area and examine the fibers carefully and note:

1. Careful examination of the fibers and branches reveals the presence of myofibrils
and cross striations.
2. The presence of intercalated disks .
3. The nuclei frequently appear to be located in a mid-fiber position.

Bone and Bone Formation

OBJECTIVES: At the end of this laboratory, you should be able to:

1. Identify the types of bone and its components

2. Identify osteoblasts, osteocytes and osteoclasts, and to describe their relationships to
each other and their role in bone remodeling.
3. Describe and differentiate bone development, including intramembranous and
endochondral ossification.
4. Describe the growth of bone include bone growth in length and width

Bone

Bone is specialized connective tissue with a calcified extracellular matrix (bone matrix) and 3
major cell types: the osteoblast, osteocyte, and osteoclast. The first type of bone formed
developmentally is primary or woven bone (immature). This immature bone is later replaced
by secondary or lamellar bone (mature). Secondary bone is further classified as two
types: trabecular bone (also called cancellous or spongy bone) and compact bone (also
called dense or cortical bone).01

Slide : Bone, femur.

The trabecular bone present in this slide is found mostly within the epiphysis and some in the
bone marrow cavity. Osteoblasts are located immediately above the osteoid (newly formed
bone matrix). Osteocytes are found within lacunae. Giant multinucleated osteoclasts, which
break down bone, are occasionally found in lacunae termed Howship's lacunae. These are
readily found in the ossification zone of the growth plate.

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The compact bone in this slide surrounds the marrow cavity and spongy bone. Locate
the periosteum (external) and endosteum (internal) linings of the bone. Note the separation of
these linings is artifact of slide preparation.

Slide : Intramembraneus ossification

Intramembranous ossification is visible in the nasal conchae on this slide. Bone arises directly
within mesenchymal condensations. This process can be identified by the appearance of bone
spicules (islands of bone) among mesenchymal cells. Look for the eosinophilic bone matrix. The
surrounding mesenchymal cells are stellate in appearance

Slide : Endochodral Ossification in Epiphyseal plate

Endochondral bone formation is represented in this slide. Bone arises by replacement of a small
hyaline cartilage model. Locate the epiphyseal plate; it is the site for bone elongation. First, find
the hyaline cartilage and move toward the bone marrow. Identify the 5 overlapping zones:

1. Zone of Reserve or Resting Cartilage - young small cells evenly distributed,

appears as typical hyaline cartilage.
2. Zone of Cell Proliferation - chondrocytes divide, forming parallel columns.
3. Zone of Cell Maturation and Hypertrophy - cells produce collagen and ground
substance
4. Zone of Cartilage Calcification - septa of cartilage matrix become calcified, cells
die.
5. Zone of Ossification - osteoblasts invade cavities, and deposit bone matrix.

Cartilage

OBJECTIVES: At the end of this laboratory, you should be able to:

1. Recognize and differentiate the three types of cartilage, the cellular and extracellular
components of there.

2. Identify the structures within cartilage: perichondrium, interterritorial matrix, capsule, territorial
matrix, isogenous groups, chondroblasts, chondrocytes and lacunae.

SLIDES FOR THIS LABORATORY :

Slide …Trachea (hyaline cartilage), embryonal tissue

Hyaline cartilage is the most common type of cartilage. Find the surrounding capsule-
like perichondrium . The chondrocytes , shrunken with tissue preparation, are located
within lacunae. The area immediately adjacent to the cell is called the capsule and stains
intensely because of the rich glycosaminoglycan content. Clusters of cells arising from mitosis
are called isogenous groups . Hyaline cartilage is easily distinguished from elastic and
fibrocartilage because of the homogeneous (no fibers are visible) extracellular matrix (ECM) .
The darker staining matrix immediately surrounding the capsule is called territorial matrix , while
the matrix that occupies the majority of space between chondrocytes is interterritorial matrix .

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Slide : Epiglottis. (elastic cartilage)

Elastic cartilage (like hyaline cartilage) has chondrocytes located in lacunae and the tissue is
surrounded by a perichondrium . Elastic fibers predominate in the ECM. The abundance of
elastic fibers gives the ECM a heterogeneous (fibrillar) appearance when stained with H & E.

Slide : Intervertebral disk.(Fibrocartilage)

Fibrocartilage is intermediate in appearance between dense connective tissue and hyaline

cartilage. Chondrocytes are situated in lacunae and no identifiable perichondrium is present.
The matrix is acidophilic because of the large amount of coarse type I collagen fibers . Note that
there are relatively few cells when compared with hyaline cartilage.

BCS 3 CARDIOVASCULAR SYSTEM, BLOOD CELLS AND LYMPHOID TISSUE

Cardiovascular system

OBJECTIVES: At the end of this laboratory session, you should be able to:

1. Identify and describe the light and electron microscopic structure of the various types of
blood and lymphatic vessels.
2. Describe the structure modifications in the three types of capillaries, and to relate this to
differences in permeability.
3. Identify the layers of the atrial and ventricular walls and understand how differences in
the thickness of these layers contribute to heart function.
4. Identify and describe the microscopic structure of a heart valve
5. Identify the components of the cardiac skeleton and understand its structural and
functional relationship with the musculature of the heart.

SLIDES FOR THIS LABORATORY:

Vessels

Blood vessels are usually composed of three layers: the tunica intima, tunica media, and tunica
adventitia. The tunica intima consists of a layer of endothelial cells lining the lumen of the
vessel, as well as a subendothelial layer made up of mostly loose connective tissue. Often, the
internal elastic lamina separates the tunica intima from the tunica media. The tunica media is
composed chiefly of circumferentially arranged smooth muscle cells. Again, the external elastic
lamina often separates the tunica media from the tunica adventitia. Finally, the tunica adventitia

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is primarily composed of loose connective tissue made up of fibroblasts and associated collagen
fibers.

Tunica intima Tunica media Tunica adventitia

1. Endothelial cell lining 1. Smooth muscle cells, collagen 1. Mostly collagenous fibers
fibers, and ground substance
2. Subendothelial layer 2. Elastic fibers (not lamellae)
2. Elastin in the form of
3. Internal elastic lamina fenestrated elastic lamellae (esp 3. Fibroblasts and
elastic arteries) macrophages

3. External elastic lamina 4. Vasa vasorum

Elastic arteries

Slide : Elastic artery (aorta).

In the aorta, the tunica media constitutes the greater part of the vessel wall. It is made up of
multiple elastic lamellae alternating with thin layers of circularly oriented smooth muscle. The
boundary between the tunica intima and media is not readily defined, and the internal elastic
lamina is merely the innermost of the many elastic lamellae within the wall. The tunica
adventitia essentially lacks elastic lamellae, and is mainly loose connective tissue and blood
vessels (vasa vasorum).

Muscular (distributing) arteries

In muscular arteries, smooth muscle becomes the predominant constituent of the tunica media.
Internal and external elastic laminae are prominent. The tunica intima is thinner than the intima
of elastic arteries.

ARTERIOLES

In arterioles, the tunica intima consists of a continuous endothelium and a very thin
subendothelial layer. In addition, a thin, fenestrated internal elastic lamina is present in larger
arterioles but absent in terminal arterioles. Generally, the tunica media consists of 2 layers of
smooth muscle cells. But in the smallest arterioles there is a single layer. The tunica adventitia
is a thin sheath of connective tissue, which is not easily defined.

Capillaries & venules

Capillaries average from 9 to 12 µ m in diameter, just large enough to permit passage of cellular
components of blood. The wall consists of extremely attenuated endothelial cells. In cross
section, the lumen of small capillaries may be encircled by a single endothelial cell, while larger

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capillaries may be made up of portions of 2 or 3 cells. No smooth muscle is present. Recall that
at the EM level, 3 types of capillaries can be distinguished: continuous, fenestrated, and
discontinuous.

Capillaries and venules are the principal vessels where exchange between the blood vascular
space and the interstitium takes place. A surprising amount of exchange of fluid and cells takes
place through the wall of the venule. Venules are similar to capillaries in that they have a very
thin wall; but unlike capillaries they have diameters that range from approximately 20 - 60 µm.

Veins

Veins are best studied along with their corresponding arteries. Veins have the same 3 layers as
arteries, but boundaries are indistinct and elastic components are not as well developed in
veins. Histologically, veins are usually collapsed because of the thinner, less elastic walls.

Slide 80

Use this slide to appreciate the contrasting features of a medium sized artery (muscular artery)
and vein . The vein is typical of vessels not surrounded and supported by solid tissue. Veins in
these locations have a well developed muscular adventitia (muscle cells are oriented
longitudinally). This is also the typical structure of large veins such as the vena cava.

Lymphatics

Lymphatics are found in all tissues except the CNS, cartilage, bone and bone marrow, thymus,
teeth, and placenta. Lymphatics are sometimes difficult to demonstrate satisfactorily in normal
tissues because these large, thin-walled vessels frequently collapse to the point of invisibility
during tissue processing. Lymphatics are often most easily visualized and studied during
pathological processes (e.g., inflammation). Lymphatics start as blind-ended lymphatic
capillaries which coalesce to form lymphatic vessels and finally empty into the circulation via the
lymphatic ducts (thoracic and right lymphatic).

BLOOD

OBJECTIVES: At the end of this laboratory session you should be able to identify the cellular
and formed elements in peripheral blood and know their functions.

PRETEST : Before beginning this laboratory, be certain that you understand the use of the
terms:

SLIDE FOR THIS LABORATORY: 22.

BLOOD COMPONENTS

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Find a region in the blood smear slide where the RBC's are separated (not overlapping). When
studying the blood smear slide, pay close attention to nuclear characteristics, staining patterns,
and cell diameter (remember that erythrocytes are usually 7-8 µm in diameter). Use the oil
immersion lens to identify each cell type.

Slide 22 Peripheral blood smear, Wright's stain.

Identify the following components:

Erythrocytes

Platelets (small blue-purple clusters)

LEUKOCYTES

Leukocytes (or White Blood Cells) can be divided into two main types: Agranular & Granular.
When studying the WBC's, keep in mind the percentages listed below which represent the
normal adult leukocyte differential.

Neutrophils Eosinophils Basophils Lymphocytes Monocytes

50-70% 2-4% 0-1% 20-40% 2-9%

AGRANULAR LEUKOCYTES

Monocytes

Look for fibrillar chromatin in the nucleus and abundant blue cytoplasm. Recall that
macrophages arise from monocytes.

Lymphocytes

Look for clumped chromatin and a thin rim of basophilic cytoplasm.

GRANULAR LEUKOCYTES

Neutrophils

These are the most common WBC. Look for multilobed nuclei, individual granules are not
distinguishable. The cytoplasm is lightly stained a "sandy" color.

Eosinophils

Look for bilobed nuclei, individual granules are distinguishable and eosinophilic (stain intensely
red-orange).

Basophils

These cells are very rare in the blood. Basophils have coarse, basophilic granules that usually
obscure the nucleus.

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LYMPHOID ORGANS

OBJECTIVES: At the end of this laboratory session, you should be able to:

1. Identify and differentiate among lymphoid organs based on their histological structure
2. Identify and understand the cells and connective tissue element of lymphoid organ

SLIDE FOR THIS LABORATORY: 23, 24, 25, 26

Slide 23: Tymus (H & E stain,magnification 100x ) :

tymus is the other primary lymphoid organ, an encapsulated, bilateral organ in the mediastinum,
is subdivided by connective tissue (see; reticular tissue) into connected lobes. Each lobes
divided in to peripheral region (cortex), where lymphocytes are fairly dense and central region
(medulla) with fewer lymphocytes. In medulla also found degeneration form of type 6 epithelial
reticular which is known as Hassal bodies.

Slide 24 : Lymph node ( H & E stain, magnification 100x) :

lymph node is one of secondary lymphoid organ, distributed throughout the body. Lymph node
show three functional regions : the cortex (C) paracortex (P) and Medulla (M). connective
tissue( reticular connective tissue) of the capsule completely surround each lymph node and
extend as several trabeculae. Cortex characterize by many lymphoid nodules, whether in
Medulla we found medulary sinuses and cords . Paracortex is an area between cortex and
medulla. Lymphoid nodule composed of lymphocytes with reticular connective tissue.

Slide 25 : Spleen (H & E stain, magnification 100x) :

Spleen is largest lymphoid organ. Spleen located in left abdomen quadrant. Spleen is
composed of reticular tissue containing lymphocytes and other blood cells. The spleenic pulp
has two component , the white pulp and the red pulp. The white pulp consist of lymphoid tissue
surrounding the central arteoles as the periarteriolar lymphoid sheath. The red pulp consist of
spleenic sinusoid (contain blood cells of all types) and spleenic cord ( reticular tissue rich in
lymphocytes).

Slide 26 : Palatine Tonsila (H & E stain, magnification 100x).

Palatine Tonsils are partially encapsulated lymphoid tissue lying beneath and in contact with
the epithelium of the oral cavity. Palatine tonsils in the posterior parts of the soft palate, are
covered by stratified squamous epithelium. Each has 10-20 epithelial invaginations that
penetrate the tonsil deeply, forming crypts. In tonsils we found masses of lymphoid nodule
consist of reticular tissue, rich in lymphocytes.

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BCS 4 RESPIRATORY SYSTEM

OBJECTIVES: When finished with this laboratory, you should be able to:

1. Discuss and identify the conducting and respiratory portions of the respiratory tract.
2. Identify the morphological changes and their functional significance in the walls of the air
passages.
3. Identify and discuss the specific functions of the epithelial cells covering the entire
respiratory tract.

SLIDES FOR THIS LABORATORY:

Slide 11 Nasal Mucosa

Find typical respiratory epithelium of the nasal mucosa and the modified epithelium that
comprises the olfactory epithelium . Compare the respiratory epithelium with the olfactory
epithelium. Note the large nerve bundles beneath the olfactory epithelium that eventually will
make their way to the olfactory bulbs via the cribriform plate. These are called filia
olfactoria (axons of bipolar olfactory neurons). Note also the Bowman's glands in the mucosa.

Slide 13 Trachea.

The trachea is a membranous tube with walls that are supported by incomplete cartilaginous
rings . Note that these rings open toward the esophagus. Observe the mucous membrane
of ciliated pseudostratified columnar epithelium with goblet cells. This epithelium has an
unusually well defined basement membrane . The wall of the trachea is composed of a mucosa,
submucosa, cartilaginous layer, and adventitia. The lamina propria of the mucosa contains
many elastic fibers, lymphoid tissue in diffuse patches, and occasional small nodules. In
addition, mucous glands with serous demilunes are present in the submucosa. Follow the
cartilage rings around to the free ends and observe the trachealis muscle . This muscle is
smooth muscle and attaches to the free ends of the cartilage rings. Some specimens may
contain small portions of the thyroid gland anteriorly.

Slide 14 Lung.

Airways

First identify a bronchus . The trachea first branches into two primary bronchi and then each
bronchus branches several times. The characteristic feature of the bronchus is the presence
of plates of hyaline cartilage . Smooth muscle and mucus secreting cells (Goblet cells) are also
present. The bronchus is lined by ciliated pseoudostratified columnar epithelium . Next find
a terminal bronchiole . These air passages are characterized by the absence of alveoli and the
presence of cartilage and goblet cells. Smooth muscle is still evident within the muscularis
mucosae . Now observe a respiratory bronchiole , this is the first structure where gas exchange
occurs. Look for the presence of alveoli , and recall the presence of just one alveolus is enough
to classify a respiratory bronchiole. The wall of a respiratory bronchiole may be cuboidal
epithelium . Next, locate an alveolar duct which are seen as elongate air spaces with almost no

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walls. An alveolar sac is composed of clusters of alveoli. Finally, the alveolus is a single sac-like
air space.

Cells

Using the oil immersion lens, focus on an alveolar wall. Observe the alveolar capillaries which
are identified by erythrocytes in the lumen. Note the squamous epithelial cells that form part of
the alveolar wall. These are Type I Pneumocyte cells . Also find Type II Pneumocyte cells which
can be seen as cells that bulge into the alveolus. Finally, alveolar macrophages (dust cells) can
be easily located in the air space of alveolar sacs.

Visceral pleura

Find this connective tissue layer at the outer margin of the lung.

BCS 5 NERVOUS SYSTEM

Nervous Tissue

OBJECTIVES: At the end of this laboratory session, you should be able to:

1. Identify neurons and understand the functional relationship between the soma and cell
processes.

2. Distinguish between glial cells and neurons and know the different functions of these
supporting glial cells.

3. Identify and understand the connective tissue elements of the central and peripheral nervous
systems.

4. Distinguish between specific organs/structures in the central and peripheral nervous systems
(i.e. cerebellum, peripheral nerve, autonomic ganglion).

SLIDES FOR THIS LABORATORY: 50, 51, 52 and 53

Slide 50 Spinal cord.

Note the H-shaped central gray matter (containing many neurons) and the surrounding white
matter . The gray matter is organized as posterior (dorsal) horns and anterior (ventral) horns .
Also note the pia mater that is closely adherent to the spinal cord. Identify the ependymal
cells surrounding the central canal . Observe the large neuron cell bodies (somata or perikarya)
in the ventral horn and the much smaller cell bodies of the dorsal horns. Identify the neural
processes ( axon and dendrites ) and axon hillock of a large multipolar motoneuron. Observe
the large number of nuclei of other (non-neuronal) cells within both gray and white matter.
Although it is not possible to identify each cell type with certainty on this routine-stained section,
most of these represent the nuclei of neuroglial cells (astrocytes, oligodendroglia, and

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microglia). Examine the white matter at high magnification and note nerve cell
processes surrounded by empty myelin spaces and the nuclei of neuroglial cells.

Slide 51 Cerebellum and medulla.

In the cerebellum, the gray matter consists of an inner granular layer , Purkinje cell layer ,
and outer molecular layer . Identify the large Purkinje cells of the Purkinje cell layer. These three
layers (molecular, Purkinje, and granular) constitute the cerebellar cortex . Observe the
numerous granule cells (small neurons) constituting the granular layer. Note the pia
mater closely adherent to the cerebellum. Note the differences between the cerebellum and
medullawhich are both present in this slide. Locate the small amount of choroid plexus between
the medulla and cerebellum. Observe the cuboidal appearance of the modified ependymal
cells that make up the choroidal epithelium.

Slide 52 Cerebrum.

Most external in this slide is the pia mater , then gray matter , and most internally is the white
matter . The gray matter is arranged in six ill-defined layers of nerve cells . See if you can
delineate the layers of cells in the cortex (you will not be required to do this for exam purposes).
Study the larger neurons in the slide; they are pyramidal cells and are a type of multipolar
neuron. Observe the large apical dendrite projecting toward the surface of the cortex. Observe
the many nuclei of glial cells .

PERIPHERAL NERVOUS SYSTEM

Slide 53 Peripheral nerve.

In the cross section, note that the peripheral nerve contains at least two nerve
fasicles separated from each other by connective tissue containing fat cells. Identify
epineureum, perineureum, and endoneurium. In the longitudinal section, notice the regular
parallel orientation of myelinated nerve cell processes and connective tissue in longitudinal
section. At high magnification, note the elongate nuclei of Schwann cells . Also
find axons , myelin sheaths (of Schwann), and nodes of Ranvier .

BCS 6 UROGENITAL SYSTEM

URINARY SYSTEM

OBJECTIVES : At the end of this laboratory you should be able to:

1. Identify the kidney, ureter, and urinary bladder microscopically.

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 2. Distinguish between the cortex and medulla of the kidney.
3. Identify the segments of the nephron, ie, the proximal tubule, distal tubule, and
Bowman's capsule with the glomerulus and the loop of Henle.
4. Identify medullary rays, distinguishing them from other regions of the kidney cortex
5. Identify the urinary and vascular poles of Bowman's capsule.
6. Correlate the light microscopy of each part of the nephron with its ultrastructure.

SLIDES FOR THIS LABORATORY :

Slide : Kidney.

Observe the cortex , medulla , and delicate connective tissue capsule . Note the relatively large
arteries and veins between the cortex and medulla. These are the arcuate arteries and veins .
Recall that arcuate arteries give rise to interlobular arteries which run through the cortex. From
the interlobular arteries, the afferent arterioles branch off to supply the kidney glomeruli.

Renal corpuscles (glomerulus + Bowman's capsule) as well as the proximal and distal
convoluted tubules are found in the cortex. One can distinguish proximal convoluted tubules
from distal convoluted tubules by these characteristics:

1) Staining - proximal tubules are more eosinophilic than the distal.

2) Nuclei - proximal tubule cells have more heterochromatic nuclei than those of distal
tubule cells; and there are more nuclei per unit area in the distal tubule than there
are in the proximal tubule.
3) Cytoplasm - the distal tubule cells appear with less cytoplasm.
4) Microvillus "brush" border - in the proximal convoluted tubules the “brush” border has
sloughed and can be seen in the lumen; thus, the lumen appears partially filled.

Observe the medullary rays and identify collecting tubules within the rays. Identify
the vascular and urinary poles of the renal corpuscles. Also observe the parietal layer
ofBowman's capsule , the urinary space , and podocytes . Find the macula densa of the distal
convoluted tubule.

Carefully study the medullary region and note the vasa recta , thin and thick limbs of the loop of
Henle , and collecting ducts . Collecting ducts in the medulla are distinguished by the clarity with
which the borders of adjacent lining cells stand out.

Supplemental Slide 111 Kidney.

In this slide, the tubules are better preserved. The proximal tubule nuclei are smaller and stain
more intensely than those of the distal tubules .

Slide : Ureter.

Observe the transitional epithelium . Examine the layers of smooth muscle . Inner longitudinal
and outer circular layers are distinguishable in this slide. Note the lamina propria . The obvious
spaces in this layer are artifact caused by excessive shrinkage and extraction of ground
substance.

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Slide : Urinary Bladder.

Note the mucosa , submucosa , muscularis , and the serosa or peritoneal surface of the
bladder. Observe the inner longitudinal, middle circular and outer longitudinal bands ofsmooth
muscle in the muscularis. Examine the transitional epithelium with its characteristic dome
shaped surface cells. Observe any nerves or autonomic ganglion cells in the bladder wall.

BCS 7 Alimentary System and Endocrine System

Gastrointestinal tract

OBJECTIVES: At the end of this laboratory, you should be able to:

1. Identify the different papillae located on the tongue.

2. Identify and describe the regional differences in the upper GI tract
3. Identify and describe regional differences in the lower GI tract, from the beginning of the
small intestine through the anal canal.

SLIDES FOR THIS LABORATORY: 3, 40, 41, 42, and 43

Slide 40 Tongue.

This slide is representative of the posterior region of the tongue. The epithelium of the dorsal
tongue is a stratified squamous keratinized epithelium . Note the cornified layer of cells may
have been lost in preparation of this slide. Remember, most of the dorsal tongue is covered with
papillae, of which there are four principal kinds: Filiform, Fungiform, Circumvallate, and Foliate
papillae.

The posterior region is where the circumvallate papillae are located and these are present in
this slide. Observe the minor salivary glands of the lamina propria and submucosa (continuous
in the tongue). Von Ebner's glands are specialized minor serous salivary glands that drain into
the trench around the circumvallate papillae. Taste buds are embedded within the epithelium of
fungiform, foliate, and circumvallate papillae. In this slide, look for taste buds on the lateral walls
of the circumvallate papillae. Note the skeletal muscle and its relationship to the glands.

Slide 3 Esophagus.

Rely on the type of muscle in the muscularis externa to determine what region (upper, middle,
lower) of the esophagus is represented on this slide. Identify the mucosa (epithelium, lamina
propria, and muscularis mucosa),submucosa , esophageal glands , muscularis
externa and adventitia . Note the presence of small arteries and veins in the adventitia on this
slide. The larger of the veins are known as esophageal veins which may enlarge in the case of
cirrhosis of the liver to provide an alternate pathway for blood to be transported from the
intestines to the vena cava. Observe the presence of the myenteric (Auerbach's plexus) in
between the inner circular and outer longitudinal layers of the muscularis externa.

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Slide 41 Fundic stomach.

The most characteristic feature of the fundic region of the stomach is the presence of fundic
glands which contain parietal and chief cells , as well as mucous neck cells . Examine the fundic
glands and observe the surface epithelium , gastric pits , neck , and base regions of the glands.
Recall that the same type of glands are also found in the body region of the stomach. Fundic
glands are straight long, tubular glands with shallow pits. The greatest density of chief cells is in
the basal region of each gland. Thus, the lower base region appears more basophilic than the
neck and upper base region. The parietal cells are slightly eosinophilic. Observe
the mucosa , muscularis mucosae , submucosa , muscularis externa , and serosa . Also study
the submucosal and myenteric plexi of nerves and ganglia.

Slide 2 Duodenum.

Observe Brunner's glands in the submucosa. These glands are diagnostic of the duodenum.
Examine the inner circular and outer longitudinal layers of smooth muscle in the muscularis
externa . Find the peritoneal surface and note the delicate serosa . Observe the location of
the muscularis mucosae , which is often disrupted by Brunner's glands. Note the large
broad villi of the mucosa. The core of the villus is an extension of the lamina propria and also
contains a blind ending lymphatic capillary, the lacteal . Study the simple columnar
epithelium populated by goblet cells and the absorptive enterocytes with apical microvilli . The
goblet cells increase in number from the proximal to distal small intestine. The cytological
preservation in the crypts of Lieberkuhn is not very good and therefore it is difficult to observe
the many mitotic figures that are normally present. Paneth cells may be observed at the base of
the crypts but are not very well preserved. Recall that the crypts are longer in the duodenum
than in the jejunum. Study the Auerbach's (myenteric) plexi between the layers of the
muscularis externa.

Slide 42 Ileum.

The distinguishing feature of the ileum is the presence of large groups of lymphatic nodules in
the lamina propria called Peyer's patches . The plicae circulares (valves of Kerckring) in this
slide are not as prominent as those found in the duodenum and jejunum. Observe the villi and
compare to the duodenum and jejunum. Here the villi are much more leaf-like in appearance
when compared to the finger-like villi of the duodenum and jejunum. Note themuscularis
mucosa and the absence of glands in the submucosa. Examine the muscularis externa and
recall the inner circular and outer longitudinal organization. Study the crypts of Lieberkuhn and
observe paneth cells . Observe the density of goblet cells covering the villi and lining the crypts
of Lieberkuhn. Goblet cells are most numerous in the terminal ileum. Finally, observe
the serosa .

Slide 43 Colon.

There are no villi or plicae circularis in the large intestine. The simple columnar epithelium has
abundant goblet cells interspersed with absorptive cells . The crypts of Lieberkuhn are larger
than in the small intestine. This slide best demonstrates the pale staining enteroendocrine
cells (argentaffin or APUD cells) at the base of the crypts. Note the absence any glands in

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the submucosa . Examine the muscularis mucosae and the muscularis externa . Next, observe
the serosa . Find Auerbach's plexi (between the layers of the muscularis externa)
and Meissner's plexi (beneath the muscularis mucosae within the submucosa).

Liver and pancreas

OBJECTIVES: At the end of this laboratory you should be able to:

1. Recognize liver and pancreas.

2. Identify and understand the functional significance of the vascular supply and bile
drainage of the liver.
3. Identify and discuss the structural and functional arrangement of the classical hepatic
lobule, portal lobule, and the liver acinus.
4. Identify excretory ducts, interlobular ducts, intercalated ducts, acinar and centroacinar
cells in the pancreas. You should also be able to distinguish between the islets of
Langerhans and the remaining parenchyma of the pancreas.

SLIDES FOR THIS LABORATORY: 34, and 35

Liver

Slide 35 Human liver.

The principal parenchymal cell is the hepatocyte which all look very similar with H & E staining.
Some hepatocytes are binucleate . The connective tissue present is mostly seen at the border
regions between classic liver lobules . Note the connective tissue covering the uncut surfaces of
this section. This is Glisson's capsule . Observe how the connective tissue septae dividing the
lobules are continuous with the capsule

You can easily observe the components of the classic lobule on this slide. Find central
veins and identify the plates (or cords) of hepatic cells that radiate out from the central vein.
These cords of hepatic cells are separated bysinusoids . At the 6 apices of the classic lobule,
observe the portal triads made up of hepatic (portal) artery, portal vein, bile duct, and lymphatic
vessels. The lymphatic vessels are not visible in this preparation. Also locatesublobular
veins which are found separate from the hepatic (portal) artery, portal vein, and bile duct.
Sublobular veins coalesce into the hepatic vein which carries blood from the liver to the vena
cava.

Exocrine pancreas

Slide 34 . Pancreas.

The exocrine component of the pancreas is a compound tubuloacinar gland with a complex
series of ducts that convey the exocrine secretion of the pancreas to the duodenum. Observe
the serous acini of the pancreas. Each acinus is composed of acinar cells . Note the
intense basophilia in the basal portion of the acinar cells and eosinophilia of the apical portion.

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Some acini reveal the centrally located centroacinar cells . These cells are the beginning
of intercalated ducts lined with a simple low columnar or cuboidal epithelium. The intercalated
ducts empty into the intralobular ducts which have a cuboidal epithelium. These ducts empty
into the interlobular ducts which run within the connective tissue septa dividing the pancreas into
lobules. The interlobular ducts coalesce to form the pancreatic duct which empties into the
duodenum.

The endocrine portion of the pancreas, the islets of Langerhans , are seen as islands of pale
staining cells spread through the organ. Defer your detail study of these until the endocrine
glands are covered in a later laboratory.

Note: the pancreas can be distinguished from the parotid gland in that the pancreas does not
have striated ducts, and the pancreas has the characteristic islets of Langerhans.

BCS ENDOCRINE ORGANS

OBJECTIVES: At the end of this laboratory, you should be able to:

1. Identify the pars distalis, pars intermedia, and pars nervosa; and recognize the individual
cell types in the pars distalis.
2. Identify the thyroid gland and the specific cell types and their functions.
3. Identify the parathyroid gland and the specific cell types and their functions.
4. Identify the adrenal gland, distinguish cortex from medulla, distinguish the three zones of the
cortex, and identify specific cell types within the adrenal gland and give their function.
5. Identify islets of Langerhans in the pancreas and distinguish them from the exocrine portion
of the gland, name the cells types present within each islet and give their function.

SLIDES FOR THIS LABORATORY: 30, 31, 32, 33, and 34

PITUITARY GLAND

Slide 30 Pituitary gland.

PARS DISTALIS . The pars distalis is composed of two general cell types: chromophils (50%)
and chromophobes (50%). The chromophils can be further subdivided into acidophils (40%)
and basophils (10%). The acidophils secrete GH (somatotropes) and prolactin (mammotropes).
Basophils secrete TSH (thyrotropes), LH (gonadotropes), FSH (gonadotropes), and ACTH
(corticotropes). The different acidophils and basophils cannot be distinguished in the light
microscope. Chromophobes are undifferentiated or resting chromophils that appear weakly
stained with smaller nuclei and less distinct borders. Observe the numerous blood vessels , the
delicate connective tissue framework , and the connective tissue capsule . Recall that the
hypophyseal portal circulation carries releasing hormones from the hypothalamus to the
adenohypophysis targeting the acidophils and basophils and causing release of hormones into
the blood stream.

PARS NERVOSA . Nerve fibers fill most of the pars nervosa but they are not easily identifiable
without special stains. Note that the main cell type here is a glial or supporting cell called

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a pituicyte . The bulk of the pars nervosa consists of axons from neurons in the supraoptic and
paraventricular nuclei of the hypothalamus.

PARS INTERMEDIA . This structure (rudimentary in humans) lies between the pars distalis and
pars nervosa. It consists mainly of colloid filled cysts lined by cuboidal epithelium. Note:
a lumen may be present between the pars intermedia and pars distalis.

THYROID GLAND

Slide 31 Thyroid gland.

The thyroid follicle forms the parenchymal unit of the thyroid gland. Each follicle consists
of follicular epithelium (simple cuboidal) and a central mass, the colloid . Active follicles
generally have "high" epithelium. Each follicle is surrounded by a delicate layer of loose
connective tissue , containing a capillary network. Thus, the follicle cells are bounded by colloid
on one side, and are in association with capillaries on the other. Try to identify parafollicular
cells or C cells which are located within the confines of the follicular basement lamina yet
excluded from the follicle lumen by follicular epithelium cells. These cells secrete calcitonin and
may be difficult to identify. They are somewhat larger and lighter staining than follicular cells.

PARATHYROID GLAND

Slide 32 Parathyroid gland.

The parathyroid gland, like all endocrine glands, has a capsule and is well vascularized. Note
the general arrangement of solid cell cords that form the gland. Note the clear-cut cell outlines
of principal (chief) cells which have a slightly eosinophilic cytoplasm containing lipofuscin
pigment granules and moderate amounts of glycogen. Principal cells produce parathyroid
hormone (PTH). Identify oxyphil cells which are larger and very eosinophilic. The function of
oxyphil cells is unknown but they may also secrete PTH. Adipose tissue is frequently found in
this gland in older individuals.

ADRENAL GLAND

Slide 33 Adrenal gland.

Examine both the cortex and the medulla . The human adrenal gland resembles a flattened
triangle in cross section but frequently may be very irregular in shape due to the presence of
deep folds involving the entire organ.

ADRENAL CORTEX . The cortex is divided into three zones (starting from the capsule): the
zona glomerulosa, the zona fasciculata, and the zona reticularis. Identify the capsule and study
the cell arrangement in the various zones. The zona glomerulosa is composed of cells grouped
in an ovoid configuration. The zona fasciculata is composed of lipid-droplet laden cells arranged
in radial columns. Finally, the zona reticularis is composed of a loose meshwork configuration.
Know the major classes of secretions produced by each morphological area.

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Zone Secretion

Zona Glomerulosa Mineralocorticoids (aldosterone)

Zona Fasciculata Glucocorticoids (cortisol)

Zona Reticularis Weak androgens (DHEA)

ADRENAL MEDULLA . The medulla is entirely surrounded by the cortex. The principal cells are
the chromaffin cells which can be thought of as modified post-ganglionic sympathetic neurons
that lack dendrites and axons. Identify the central vein , noting particularly the abundance and
arrangement of smooth muscle in the wall.

ISLETS OF LANGERHANS

Slide 34 Pancreas.

Within the parenchyma of the pancreas you will note small islets or clusters of lighter staining
cells - these are the islets of Langerhans (endocrine part of the pancreas). Note the number of
islets on your sections and the variation in their size. The islets are composed of three cell
types: alpha cells, beta cells, and delta cells. These cells cannot be readily distinguished without
special stains. Alpha cells (15-20%) are found at the periphery and secrete glucagon. Beta cells
(70%) are found more centrally and secrete insulin. Delta cells (5-10%) are found throughout
the islet and secrete somatostatin.

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