1.

Introduction to Cells, Undergraduate – Graduate

Histology Lecture Series

Tissues, and Microscopy Larry Johnson, Professor

Veterinary Integrative Biosciences
Texas A&M University
College Station, TX 77843
VIBS 443 and VIBS 602
 OBJECTIVES

1. Preview cellular ultrastructure

2. Preview cells, tissues, and organs

3. Overview of light and electron microscopy

4. Preparation of specimens – types of visions

5. Section orientation
 Introduction to HISTOLOGY
PROTOPLASM – Living Substance
CELL – Smallest unit of protoplasm
CELL Simplest animals consist of a single cell.

TISSUE – Groups of cells with same general function

and texture (texture = tissue)
TISSUE
e.g., muscle, nerve, connective tissue, epithelium
ORGAN – Two or more types of tissues;
larger functional unit
ORGAN
e.g., skin, kidney, intestine, blood vessels
ORGAN SYSTEM - Several organs
e.g., respiratory, digestive, reproductive systems
SYSTEM
 tissue

organ
cell

organ system
 In the 1660s, Robert Hooke looked
through a primitive microscope at
a thinly cut piece of cork. He saw a
series of walled boxes that
Onion Root Mitosis reminded him of the tiny rooms, or
cellula, occupied by monks, and
he coined the word "cell.“
Cells
in a 
plant
Although animal cells do not
have cell walls like plants,
cells are the “building blocks
of life” of both.
 FOUR BASIC TYPES OF
TISSUES IN THE BODY
-----------------------------------------------
Epithelium Connective tissue

Muscular tissue Nervous tissue

 Epithelium
Functions:
Cover organs, line viscera and blood
vessels, secretory cells of glands
 Epithelium
Distinguishing features and
distribution:
• Always sit on a basement membrane, but
come in a variety of configurations:
classified on the basis of their shape and
of the surface cells and whether one
(simple) or more (stratified) layers of
cells are stacked upon each other.
• These cells are always attached to their
neighbors by desmosomes, tight
junctions, and gap junctions.
 Epithelium
Histological Identification

Simple Squamous –
single layer of flat cells
(blood vessels, covering of
organs)
Stratified Squamous –
Multiple layers of cells with
flat ones at the surface
(skin, gums)
 Epithelium
Histological Identification

Simple cuboidal – Single

layer of square cells
(kidney tubules, liver
cells, many others)

Simple columnar –
Single layer of tall, thin
cells (intestinal
epithelium)
 Epithelium
Histological Identification
Pseudostratified columnar – single
layer of tall, thin cells packed together
in such a jumble that they seem to be
in layers, although all of the cells reach
the basement membrane (respiratory
passage)

Transitional – stratified cuboidal

epithelium of urinary passages
 Epithelium
Histological Identification
Some epithelia have surface
specializations such as
numerous microvilli or cilia.
EPITHELIUM

CONNECTIVE
TISSUE

Skin

Gunther von Hagens’

BODY WORLDS
 Connective Tissue

Function:
the histological glue which binds the other
tissues together to form organs,
specializations include blood, cartilage, and
bone.

CONNECTIVE
TISSUE
 Connective Tissue
Distinguishing features and histological identification:
• Loose connective tissue – sparse collagen and elastic
fibers, plentiful cells including fibroblasts, leukocytes

• Dense connective tissue – concentrated collagen, few cell

• Cartilage – avascular homogeneous matrix of collagen and

protein-polysaccharides with few cell

• Bone – calcified collagen matrix with few cells trapped in

the caves of bone
CONNECTIVE
TISSUE
 Fat
Muscle

Gunther von Hagens’ Body Worlds

 Muscle
Function:
generation of contractile force.

Distinguishing features:
• High concentration of contractile
proteins actin and myosin arranged
either diffusely in the cytoplasm
(smooth muscle) or in regular
repeating units called sarcomeres
(striated muscles, e.g., cardiac and
skeletal muscles).
 Muscle
Histological identification:
• Skeletal muscle – very long cylindrical
striated muscle cells with multiple
peripheral nuclei
• Cardiac muscle – short branching
striated muscle cells with one or two
centrally located nuclei
• Smooth muscle – closely
packed spindle-shaped
cells with a single centrally
placed nucleus and
cytoplasm that appears
homogeneous by light
microscopy
 Muscle
Distribution:
• Skeletal – striated muscles mostly
associated with the skeleton
• Cardiac – striated muscles associated with
the heart
• Smooth – fusiform cells associated with the
viscera, respiratory tract, blood vessels,
uterus, etc.
 Smooth muscle

NERVOUS
TISSUE

Gunther von Hagens’

Body Worlds
Nervous Tissue
Functions:
specialized for the transmission,
reception, and integration of
electrical impulses

Distinguishing features:
• neurons – very large excitable cells with long
processes called axons and dendrites. The axons
make contact with other neurons or muscle cells at
a specialization called a synapse where the
impulses are either electrically or chemically
transmitted to other neurons or various target cells
(e.g., muscle). Others secrete hormones.
Nervous Tissue

Bipolar
neurons
of inner ear
 Nervous Tissue
Glial cells – the supporting cells of nervous tissue.
Nerves – collections of neuronal processes bound
together by connective tissue. Axons may be
coated by a myelin sheath (“myelinated”) or
simply protected by being cradled in an
indentation of a glial cell (“unmyelinated”).

Myelinated

Unmyelinated?
 Nervous Tissue
Nerves – collections of neuronal processes bound together by
connective tissue. Axons may be coated by a myelin sheath
(“myelinated”)

DEMO SLIDE BOX 78

Nervous Tissue
Distribution:
• Comprise the central nervous
system
• Individual peripheral nerves
are found throughout the
body
• Individual neurons and
clusters of neurons (called
ganglia) are found in most
organs
Gunther von Hagens’
Body Worlds
 Blood cells -functions
(classified as connective tissue)
Functions:
• Red cells – carry oxygen to the tissues
and CO2 from tissues
• White cells – transient inhabitants of the
blood which are manufactured in bone
marrow and pass through the blood to
connective tissue where they
participate in defense against biological
and chemical invaders
• Platelets – blood clotting
 Blood cells
(classified as connective tissue)

Distinguishing features and histological

identification:
• red cells – biconcave discs containing
hemoglobin
• white cells – granulocytes have one of three
different types of granules and lobed
nuclei; lymphocytes and monocytes have
few granules and round or indented
nuclei
• platelets – anucleate cell fragments
produced by megakaryocytes in the bone
marrow
 LYMPHOID TISSUE
Functions:
responsible for the “immune response” to foreign
invaders which is mediated by either antibody
produced by the cells or by the cells themselves.

Amplification of the immune response:

factors (lymphokines) and other cell types
involved
 Lymphoid Tissue
Distinguishing features and
histological identification: clusters
of lymphocytes free in the connective
tissue or surrounded by a connective
tissue capsule as in a lymph node

Distribution:
in primary lymphoid organs (spleen,
thymus, lymph nodes); along alimentary
canal and respiratory passages.
Blood Vessels
Not one of the four basic tissues, but necessary to learn at this
stage; All blood and lymph vessels are lined with endothelium
(a simple squamous epithelium)
 Blood Vessels
Histological identification:
• Artery – thick wall composed of
smooth muscle plus some connective
tissue surrounding a small lumen
• Capillary – narrow tube lined with a
single endothelial cell
• Vein – large lumen relative to
thickness of connective tissue and
smooth muscle wall
• Lymphatic – small thin walled vessels
which carry lymph
Gunther von Hagens’
BODY WORLDS
 Where are these basic tissues located?
EPITHELIUM
CONNECTIVE TISSUE
MUSCULAR TISSUE
NERVOUS TISSUE

Epithelium
 Where are these basic tissues located?
EPITHELIUM
CONNECTIVE TISSUE
MUSCULAR TISSUE
NERVOUS TISSUE

Connective
tissue
 Where are these basic tissues located?
EPITHELIUM
CONNECTIVE TISSUE
MUSCULAR TISSUE
NERVOUS TISSUE

Muscular
tissue
Where are these basic tissues located?
EPITHELIUM
CONNECTIVE TISSUE
MUSCULAR TISSUE
NERVOUS TISSUE

NERVOUS
TISSUE
 Magnification vs. Resolution
1. Magnification - increase in image size
2. Resolution - smallest distance between two
points that can be seen (distinguished)
 Magnification vs. Resolution

1. magnification - increase in image size

2. resolution - smallest distance between two
points that can be seen (distinguished)

Calculated by:
0.61 (wavelength) / numerical aperture

0.25 um for light microscope

0.1 nm for electron microscope
 Sample Preparation
1. Fixation
2. Embedding
A. Paraffin
B. Plastic
 Sample Preparation
1. Fixation
2. Embedding
A. Paraffin
B. Plastic
3. Sectioning
A. 0.5 um for Light Microscopy
B. 60-80 nm for Electron Microscopy
Sample Preparation
 Staining
1. Light Microscopy
A. Hematoxylin and Eosin (H&E)
B. Periodic Acid/Shiff (PAS)
C. Toluidine Blue

2. Electron Microscopy
(TEM)
A. Osmium
B. Lead Citrate
 Staining
1. Light Microscopy
A. Hematoxylin and eosin (H&E)

B. Periodic Acid/Shiff (PAS)

Color provides clues

C. Toluidine Blue

Shape
Size
Intensity of staining
 Staining
1. Light Microscopy
A. Hematoxylin and Eosin (H&E)
B. Periodic Acid/Shiff (PAS)
C. Toluidine Blue

2. Electron Microscopy
(TEM)
A. Osmium
B. Lead Citrate
145 Fundic stomach (H&E)
243 Fundic stomach, monkey (PAS)
244 Surface mucus cells of Fundic stomach,
rabbit (toluidine blue)
 Toluidine Blue
Eosin (H&E) of retina
19709 Transparency of unstained tissue
Phase Contrast
Dead stained cells
Bright Field

Live unstained cells

Phase Contrast

Nomarksi
“differential interference
contrast”

Dark Field
 Magnetic
lens

Glass
lens
Other Light and Electron Microscope
Procedures

• Immunofluorescence
• Autoradiography
• In Situ Hybridization (ISH)
• Freeze Fracture
– (Membrane Analysis)
AUTORADIOGRAPHY
self radioactive
Evidence for protein pathway

Autoradiography
Procedure to localize a
product (e.g., protein) within a
cell or gel that is self-
radioactive due to the cell’s
incorporation of radioactive
precursors (e.g., radioactive
amino acids) into that product
that is visualized in a
photographic emulsion.
Autoradiography vs Fluorography
Terminology
Autoradiography is the direct exposure of
film by beta particles or gamma rays.

Fluorography is the exposure of film by

secondary light that was generated by the
excitation of a fluor or a screen by a beta
particle or a gamma ray.
 In situ hybridization
• In situ hybridization (ISH) is a type of
hybridization that uses a labeled
complementary DNA or RNA strand (i.e.,
probe) to localize a specific DNA or RNA
sequence in a portion or section of tissue (
in situ).
Source: http://en.wikipedia.org/wiki/In_
situ_hybridization
Photomicrograph of a
section of human epithelial
tumor (condyloma) in which in
situ hybridization with the DNA
of the human papilloma virus
type II (HPVII) was performed.
Observe dark staining of
several nuclei, indicating the
presence of the genome of
this virus in the tumor,
suggesting its possible
participation in the genesis of
the tumor.
 Carbon Replica
1. Heavy metal evaporated from
a filament “shadows” the
specimen.

2. A strengthening film of
carbon evaporated from above.

3. The replica is floated onto the

surface of a powerful solvent to
dissolve away the specimen.

4. The replica is washed and

picked up on a copper grid for
examination.
Typical TEM

Carbon replica TEM

Section orientation

Nerve axons
Section Orientation Plane of section can make it look as if there are more
layers of cells than there are, but it can never make it
look like there are less than the actual number.

It looks like multiple

layers layer
of cells

One layer
of cells
 Plane of section can make it look as if there are more
Section Orientation layers of cells than there are, but it can never make it look
like there are less than the actual number.
Next time

Membrane and Receptors

 Many illustrations in these VIBS Histology YouTube videos were modified from
the following books and sources: Many thanks to original sources!

• Bruce Alberts, et al. 1983. Molecular Biology of the Cell. Garland Publishing, Inc., New York, NY.
• Bruce Alberts, et al. 1994. Molecular Biology of the Cell. Garland Publishing, Inc., New York, NY.
• William J. Banks, 1981. Applied Veterinary Histology. Williams and Wilkins, Los Angeles, CA.
• Hans Elias, et al. 1978. Histology and Human Microanatomy. John Wiley and Sons, New York, NY.
• Don W. Fawcett. 1986. Bloom and Fawcett. A textbook of histology. W. B. Saunders Company,
Philadelphia, PA.
• Don W. Fawcett. 1994. Bloom and Fawcett. A textbook of histology. Chapman and Hall, New York,
NY.
• Arthur W. Ham and David H. Cormack. 1979. Histology. J. S. Lippincott Company, Philadelphia, PA.
• Luis C. Junqueira, et al. 1983. Basic Histology. Lange Medical Publications, Los Altos, CA.
• L. Carlos Junqueira, et al. 1995. Basic Histology. Appleton and Lange, Norwalk, CT.
• L.L. Langley, et al. 1974. Dynamic Anatomy and Physiology. McGraw-Hill Book Company, New York,
NY.
• W.W. Tuttle and Byron A. Schottelius. 1969. Textbook of Physiology. The C. V. Mosby Company, St.
Louis, MO.
• Leon Weiss. 1977. Histology Cell and Tissue Biology. Elsevier Biomedical, New York, NY.