WARNING: ALL THE MATERIALS (i.e.

, TEXT DOCUMENTS, DIGITAL

SLIDES, PHOTOMICROGRAPHS, PICTURES, VIDEOS, ETC.) THAT YOU
WILL USE IN THIS LABORATORY COURSE IN HISTOLOGY ARE
COPYRIGHTED. YOU ARE NOT ALLOWED TO DOWNLOAD, UPLOAD,
SHARE, COPY, OR EDIT ANY PART OF ANY MATERIAL, UNLESS
SPECIFICALLY GIVEN PERMISSION TO DO SO. YOU ARE HOWEVER
ALLOWED TO TAKE PHOTOGRAPHS OF THE MATERIALS AS THEY
APPEAR ON THE MONITOR, BUT ONLY FOR YOUR PERSONAL USE. YOU
CANNOT POST THE PHOTOGRAPHS IN ANY SITE ON THE INTERNET,
SEND THEM TO ANYONE BY E-MAIL OR ANY ELECTRONIC MEANS, OR
MASS PRODUCE THEM.
THIS LABORATORY IS MONITORED BY A CLASSROOM MANAGEMENT
PROGRAM THAT CAN DETECT VIOLATORS OF THE ABOVE RULE.
VIOLATORS WILL BE SUBJECTED TO DISCIPLINARY ACTION.

MODULE 5: MUSCLE TISSUE

This module on Muscle Tissue, the fourth type of fundamental tissue that you will study in
this course, will introduce you to the different types of muscle and the structural features
that differentiate them from each other.

Muscle tissue is composed of muscle cells (myocytes) that are supported and bound
together by intercellular material (connective tissue). Muscle cells are highly contractile
that is why muscle tissue is the basic tissue that is responsible for locomotion of the
individual and movement of the various parts of the body.

There are three types of muscle tissue: skeletal, smooth and cardiac that are made up of
skeletal, smooth and cardiac muscle cells (fibers), respectively. Skeletal and cardiac
muscle fibers are referred to as striated because, under LM, their cytoplasm exhibit
prominent alternating light and dark cross striations. In contrast, smooth muscle cells are
nonstriated because they do not exhibit cross striations.

Skeletal muscle tissue is organized to form organs called muscles (skeletal muscles),
many of which are named (e.g., gastrocnemius muscle). Cardiac muscle tissue, on the other
hand, comprises the myocardium of the heart that propels blood through the circulatory
system when it cotracts. Smooth muscle tissue, meanwhile, forms part of the wall of the
visceral organs and blood vessels. It is also present in the parenchyma of most other
internal organs and even the skin. Smooth muscle tissue is responsible for such activities as
peristalsis, emptying of the gallbladder and urinary bladder, vasoconstriction, contraction
of the uterus and pupillary constriction.

COMPETENCIES

At the end of the module on Muscle Tissue, given an LM histologic section and/or an
electron micrograph, the student can:

1. Distinguish muscle tissue from the other fundamental tissues.

2. Classify the muscle tissue in the specimen into skeletal, cardiac or smooth.
3. Identify and classify muscle cells.
4. Point out the different cellular components (organelles, myofibrils, etc.,) in
muscle cells.
5. Pinpoint the sarcomere and its different regions/zones.
6. Delimit a muscle fiber and its endoneurium from a muscle fascicle and its
perineurium and a muscle and its epineurium.
7. Delineate motor nerve endings and identify their parts.

PREREQUISITE KNOWLEDGE

Esteban and Gonzales’ Textbook of Histology, Fifth Edition

Chapter IV – Muscle Tissue, pages 49-62

LABORATORY SESSION ON MUSCLE TISSUE

This laboratory session has two parts. During the first part of the session you will study LM
digital slides while during the second part you will go to the INTERNET to study electron
micrographs and additional LM photomicrographs.

PART1. STUDY OF MUSCLE TISSUE IN LM DIGITAL SLIDES

In this part of the session, you will open and study a number of digital slides, each of which
contains histologic section/s of muscle tissue.

You may have to consult your textbook and atlas in the course of the session.

SLIDE SESSION

 Split the monitor and start/run Imagescope as you did during the previous lab
sessions.
 Open the digital slides listed below. The slides are in the folder labeled “Module 5.
Muscle Tissue” that is within the folder labeled “HISTOLOGY SLIDES,” which is on
your desktop.
 o Motor nerve ending
o Skeletal muscle
o Skeletal-smooth-cardiac muscle

SKELETAL MUSCLE
 On the filmstrip, click the slide labeled “Skeletal muscle” to display the slide on the
Imagescope main window.

The slide contains two specimens of skeletal muscle tissue: the one on the left shows
the muscle cells in cross section and the one on the right shows the muscle cells in
longitudinal section.

 Study the specimen where the skeletal muscle cells are shown in cross section under
low magnification.
o Note that the specimen consists of islands of eosinophilic structures that are
surrounded by narrow spaces. Each of the eosinophilic structures is a muscle
fiber and the space surrounding it is actually occupied by connective tissue
known as endomysium.
o Observe that the muscle fibers form clusters called fascicles that are separated
from each other by a wide space that contains connective tissue elements, which
comprise the perimysium.
o Notice that the muscle fascicles are bundled together by connective tissue
elements that comprise the epimysium.
o Now, examine the intercellular spaces occupied by endomysium under high
magnification:
 Note the connective tissue elements (fibers and cells) that make up the
endomysium. What types of cells and fibers comprise the endomysium?
 Find blood vessels, mostly capillaries, which abound in this area.
o Still under high magnification, examine the perimysium and epimysium.
 Note that these structures are also made up of connective tissue, albeit
denser than that that makes up the endomysium.
 Find blood vessels, which are generally bigger than those in the
endomysium.
o Now, still under high magnification, shift your attention to the skeletal muscle
cells (fibers).
 Note that skeletal muscle cells are large cells that can reach up to 1 mm
in diameter. You can use the capillaries in the intercellular area, which
have an average diameter of 7 m, as gauge to estimate the size of the
muscle cells.
 Note that the nuclei of the muscle fibers are located in the peripheral
areas of the cells.

 Shift back to low magnification. Pan the slide under low magnification and find, then
study, the specimen where the skeletal muscle cells are shown in longitudinal section.
 o Observe that the section consists of several muscle fascicles.
o Delineate the endomysium that envelops each muscle fiber and the
perimysium that separate the muscle fascicles from each other.
o Note that the muscle fibers are very long cylindrical cells that taper at both
ends.
o At higher magnifications, examine the muscle fibers (cells).
 Note that the cells are multinucleated and that their nuclei are oval and
longitudinally oriented.
 Note further that the nuclei of the muscle cells are located in the
peripheral portion of the cells, near the sarcolemma. But why do some
nuclei appear to be in the central area of the cells?
 See that the cytoplasm of the muscle cells exhibits transverse striations
of alternating light and dark bands. These are the reasons why skeletal
muscle cells are referred to as striated.
 Look at the striations more intently. Delineate the light bands, which are
called isotropic bands (I-bands) from the dark bands, which are called
anisotropic bands (A-bands).

SKELETAL, CARDIAC AND SMOOTH MUSCLE

 Click the slide labeled “Skeletal-smooth-cardiac muscle” on the filmstrip to put the
slide in the Imagescope main window.

This slide contains three specimens, as you can see in the thumbnail. From left to right,
the specimens are those of: a longitudinal section of a skeletal muscle, a section of the
muscularis externa of a segment of the digestive tract, and cardiac muscle tissue.

 Focus your attention on the skeletal muscle specimen first. The specimen contains three
fascicles that are made up of several muscle fibers each. Reinforce what you have
learned from the previous slide about the structure of skeletal muscle tissue by
studying this specimen under different magnifications and identifying the same
structures that you identified in the previous slide.

 Now, shift your attention to the specimen that contains a section of the muscularis
externa of a segment of the digestive tract.

The muscularis externa is one of the histologic layers of the digestive tract. The cells
that comprise this histologic layer are smooth muscle fibers that form two layers. In
one layer, the the muscle fibers are circularly arranged while in the other (sub)layer,
the muscle fibers are longitudinally arranged.

o Under relatively low magnification, delineate the two layers of the muscularis
externa. In this specimen, the layer of circularly arranged muscle fibers is rather
thick while the layer of longitudinally arranged muscle fibers is very thin. Note
that in the layer of longitudinally arranged muscle fibers the cells are shown in
 cross section, while in the layer of circularly arranged muscle fibers, the muscle
fibers are shown in longitudinal section.
o Study the layer where the muscle fibers are shown in cross section under
higher magnification.
 Note that in cells where the nucleus is visible, it is round to ovoid and
centrally located.
 Observe that the endomysium of the muscle fibers is indistinct because
the muscle cells are tightly packed.
Smooth muscle cells are also enveloped by endomysium and they likewise
gather together to form fascicles that are encased by perimysium.
o Reinforce your understanding of the enteric nervous system by locating the
myenteric plexus of Auerbach that is in between the two layers of the
muscularis externa.
o Now, examine the layer where the muscle fibers are shown in longitudinal
section.
 Observe that the muscle cells are spindle-shaped.
 Note that the nucleus of the muscle cells is located in the thick part of the
cells.
 Note further that there is only one nucleus per cell.

 Now, study the specimen containing cardiac muscle.

o In this slide, the cardiac muscle cells are variously oriented and hence, you will
see some in cross section, some in longitudinal section and some in oblique
section. Verify this by panning the specimen under relatively low magnification.
o Under high magnification, study an area of the specimen where the muscle fibers
are seen in cross-section.
 Note that each muscle fiber is enveloped by connective tissue elements
(endomysium).
 Note further that cardiac muscle fibers likewise gather together to form
fascicles that are encased by perimysium, but the fascicles and
perimysium are not very well delineated in this slide.
 In the intercellular areas, find capillaries.
o Now, find an area where the cardiac muscle fibers are shown in longitudinal
section. Study the area under various magnifications.
 Verify that the muscle fibers are individually enveloped by endomysium
and that the intercellular area is richly supplied with capillaries.
 Observe the alternating light (I-)and dark (A) bands (i.e., striations) in
the cytoplasm of the cardiac muscle cells. Why are these striations not as
prominent as those in skeletal muscle cells?
 Note that the muscle cells branch terminally.
 Notice that the muscle fibers are mononucleated (although there are
occasional binucleated ones) and that their nucleus is centrally located.
 Find intercalated discs that connect the cardiac muscle cells end-to-end.
What comprise these structures that are unique to cardiac muscle tissue?
MOTOR NERVE ENDINGS
 Click the slide labeled “Motor nerve endings” on the filmstrip to put the slide in the
Imagescope main window.

This slide contains a teased preparation of a skeletal muscle tissue that was specially
stained to show motor nerve endings.

 Pan the slide under relatively low magnification and:

o Find axons of somatic motor neurons
o Note that the terminal segments of the axons break up into numerous fine
branches.
 At higher magnification, find motor endplates:
o Observe that each motor endplate is formed by the bulb-like termination of an
axon (axon terminal) and the plasmalemma of muscle cell on which the axon
terminal comes to rest.
o Note that a single axon, because it breaks up terminally, is involved in the
formation of several motor endplates.

PART 2. STUDY OF MUSCLE TISSUE IN ELECTRON MICROGRAPHS AND

PHOTOMICROGRAPHS THAT ARE POSTED IN THE INTERNET

 Close ImageScope, if you haven’t done so yet.

 Go to the INTERNET by double clicking either the Internet Explorer icon or the
Mozilla Firefox icon on your desktop. Then, split the screen so the Google window gets
to occupy just the left half of the screen.
 Search for and visit Histology websites that feature LM photomicrographs and
electron micrographs of Muscle tissues. Visit several websites and study as many
relevant electron micrographs and photomicrographs as possible.
o In LM photomicrographs, identify and study motor endplates.
o In EMs, identify and study the structures and cells listed below:
 skeletal muscle fiber
 nucleus and cytoplasmic organellles
 myofibril
 sarcomere
 A-band
 I-band
 H-line
 Z-disc
 M-line
 T-tubule and terminal cisternae
 cardiac muscle
 nucleus and cytoplasmic organellles
 myofibril
  sarcomere
 A-band
 I-band
 H-line
 Z-disc
 M-line
 intercalated disc and the junctional complexes that comprise the
disc
 T-tubule and terminal cisternae
 smooth muscle
 nucleus
 cytoplasmic organelles

end of module