INTRODUCTION TO

HISTOLOGY

Created by: Dr. Diana Vera Estrada

Dr. Yisel Mi Guzmán Leguel

yisel.guzman@edu.uag.mx
Block I Fundamentals of medicine I
Syllabus Histology Block I
• Introduction to histology

• Basic Tissues.
– 1.Epithelial Tissue
– 2.Connective Tissue
– 3.Muscle Tissue
– 4.Nervous Tissue

• Cartilage and bone

• Blood and hematopoiesis
• Histology of the lymphatic system.
 Basic Bibliography for the course

1. (Recommended for all the topics)

Anthony L. Mescher. “Junqueira’s Basic
Histology Text and Atlas”, 14th edition.
Lange McGraw Hill.
Basic Bibliography for the course
Practice exercises and questions
James S. Lowe B. Stevens & Lowe's Human Histology. Fourth
Edition. Elsevier. (Clinical Key)
 Basic Bibliography for the course

Practice exercises and

questions
Gartner, Leslie P. PhD.
Textbook of
histology. 2017. 5TH
Edition.Elsevier.(Clinical Key)
Basic Bibliography for the course
Michael H. Ross. Histology and text Atlas with correlated cell and molecular
biology.6th edition. Lippincott Williams and Wilkins.
 Learning Objectives
• Know the definition of histology and its relationship with other sciences.
• Identify the different methods of study in histology
o Light microscopy
o Electron microscopy
o *Immunohistochemistry (no detailed information in this course)

• Understand the processes of preparing and viewing tissues by light and electron microscopy.
• Identify the most common stains utilized in light microscopy:
o Hematoxylin and Eosin (H&E)
o Mallory Trichrome
o Periodic Acid–Schiff Reaction (PAS)
o Wright-Giemsa Stain

• Understand the concept of eosinophilia and basophilia.

• Be able to identify in images the basic and acidic components of a tissue stained with H&E.
 HISTOLOGY

• Etymology:
Greek, histos (tissue)
logos (science)

The scientific study of the

structure of tissue and cells
• Biochemistry • Molecular biology

• Pathology is a branch of medical science that involves the

study and diagnosis of disease through the examination of
surgically removed organs, tissues (biopsy samples), bodily
fluids, and in some cases the whole body (autopsy).
Pathology
Preparation of Tissues for study

The most common

procedure used in
histologic research is
the preparation of
tissue slices or
“sections” that can be
examined visually
with transmitted light.
Light Microscopy
Tissue Preparation
• Steps required in preparing tissues for light microscopy include:
1. Fixation.
2. Dehydration and clearing.
3. Embedding.
4. Sectioning.
5. Mounting
6. Staining the sections.
1. FIXATION
Fixation refers to the treatment of the tissue with chemical agents that not only
retard the alterations of the tissue subsequent to death (or after removal from the
body) but also maintain its normal architecture.

Neutral Buffered Formalin Bouin Fluid

2. Dehydration and Clearing
• Dehydration: because a large fraction of the tissue is
composed of water.
• 50% alcohol
• 100% alcohol

• The tissue is then treated with xylene, a chemical

that is miscible both with alcohol and melted
paraffin.
Gives the tissue a translucent appearance
3. Embedding
• In order to distinguish the overlapping cells in a tissue and the
extracellular matrix from one another, the histologist must
embed the tissues in a proper medium and then slice them into
thin sections.
• The usual embedding medium is paraffin.
4. Sectioning
• Thin slices are removed from the block.
• For light microscopy, the thickness of each section is about 3 to 10
µm, and each section or a series of sections is mounted (placed) on
glass slides.
5. Mounting
• Paraffin must be solved out
(e.g toluol)
• Rehydration
• Mounting on a glass
*The slide preparation, from
tissue fixation to observation
with a light microscope, may
take from 12 hours to 2 days,
depending on the size of the
tissue.
6.Staining
Hematoxylin and eosin (H&E) is used most commonly

Most cells and extracellular

material are completely
colorless, and to be studied
microscopically tissue sections
must be stained (dyed).
Add a protective glass
coverslip on the slide with
clear adhesive.
Light Microscopy
Bright-Field Microscopy
With the bright-field microscope, stained tissue
is examined with ordinary light passing through
the preparation.
Maximal resolving power: 0.2 μm.

Permit clear images magnified 1000-1500 times.

• Objects smaller or thinner than 0.2 μm (such as a
single ribosome or cytoplasmic microfilament) cannot
be distinguished with this instrument
Light Microscopy B. Fundic Glands (270x) resolving power : 0.2 μm.

A. Mucosa of the Stomach (132x)

Gartner, Leslie P. PhD. “Textbook of histology”. 2017. 4TH Edition. Elsevier.

Electron Microscopy Transmission Electron Microscope (TEM)
• The TEM uses a beam of electrons
rather than a beam of light.
• This high resolution allows isolated
particles magnified as much as
400,000 times to be viewed in
detail.
RBC.

Enzyme-producing cells.
 Electron Microscopy
Scanning electron microscopy
• Provides a high resolution view of the surfaces
of cells, tissues, and organs.
• The surface of the specimen is first dried
and spray-coated with a very thin layer of
heavy metal (often gold-carbon) which
reflects electrons in a beam scanning the
specimen.
• Allows resolution of three-dimensional
subcellular structures.
Light Microscopy Stains

• Hematoxylin and Eosin (H&E)

• Masson Trichrome
• Periodic Acid–Schiff Reaction
(PAS)
• Wright-Giemsa Stain
• Silver or Gold Stains
• Stains for Elastin
Hematoxylin and eosin (H&E)

Hematoxylin and Eosin are the most

commonly used dyes in histology.
• A basic dye(hematoxylin)carries a net
positive charge on its colored portion.

• An acidic dye, such as eosin, carries a

net negative charge on its colored
portion.
Basic Dyes (+) (hematoxylin)
• (-) Anionic components: • The ability of such anionic
groups to react with a basic dye
• DNA; RNA (nuclei); phosphate
is called basophilia [Gr., base-
groups of nucleic acids). loving].
• Extracellular materials; the sulfate
groups of glycosaminoglycans, and
the carboxyl groups of proteins.
ACIDIC DYES (-) EOSIN • The reaction of cationic groups
with an acidic dye is called
acidophilia (Gr., acid-loving).
• (+) Cationic components:
• Ionized amino groups of proteins.
• Acid dyes stain the acidophilic
components of tissues such as
Membranes, proteins, collagen,
elastin, cytoplasmic filaments (muscle
cells). A. Hematoxylin
B. Eosin
C. H&E
 • Collagen fibers stains pink
• Muscles stains pink

Cardiac muscle cells in longitudinal section (×270).

Nuclei stains blue/dark purple
The periodic acid–Schiff (PAS) reaction
• The Schiff reagent is a bleached basic fuchsin that reacts with
aldehyde groups.
• Stains carbohydrates and carbohydrate-rich macromolecules. It is
used to demonstrate glycogen in cells, mucus in various cells and
tissues.

Periodic Acid—Schiff Reaction

•Glycogen stains deep red or magenta
Masson's trichrome

Collagen is stained bright blue

Cytoplasm of most cells

stains pink
Vagina, Masson trichrome
Suspect there is collagen

Suspect there is
mucus
Interpretation of Microscopic Sections
LONGITUDINAL SECTION CROSS SECTION
Interpretation of Microscopic Sections
 Artifacts

• Artifacts: an error in the

preparation process.
• In general, artifacts are
linked to methodology,
equipment, or reagents used
during preparation.
Bibliography:
• Anthony L. Mescher. “Junqueira’s basic Histology Text and Atlas”.
CHAPTER 1
• Histology & Its Methods of Study 14th edition. Lange Mc Graw Hill.

• Michael H. Ross. Histology and text Atlas with correlated cell and
molecular biology. Methods in histology. 6th edition. Lippincott
Williams and Wilkins. Chapter 1.
• Victor P. Eroschenko. Di Fiore’s Atlas of histology with functional
correlations.
• Histologic Methods. 11th Edition. Lippincott Williams and Wilkins.
Chapter 1.