Introduction to histology Lab

Dr.Mohammad Omran, MD
Arab American University
Faculty of Medicine
Outline

-Tissue preparation

-Stains

-Light Microscope

-Artifacts
From Biopsy to Microscopy - Tissue processing for light microscopy

https://www.youtube.com/watch?v=__D-DE0X0zk
Preparation of tissue for LM

The most routine one is paraffin section

stained with hematoxylin and eosin
(H&E)
Preparation of tissue for LM;The steps:
a. Obtaining the specimen: fresh, small pieces (less than 5mm3) – tissue block
b. Fixation: fixatives: use formalin or Bouin’s to preserve structural organization
c. Dehydration: use ethyl alcohol to get rid of water of tissue and cell
d. Clearing: use xylene to get rid of alcohol
e. Infiltration: The tissue is then placed in melted paraffin until it becomes
completely infiltrated with this substance.
f. Embedding: The paraffin-infiltrated tissue is placed in a small mold with melted
paraffin and allowed to harden.
g. Trimming: The resulting paraffin block is trimmed to expose the tissue for
sectioning (slicing) on a microtome.
h. Then put them on glass slides.
Preparation of tissue for LM; Stains:
Hematoxylin & Eosin (H&E) staining
--- Hematoxylin: basic stain, combines with acidic
components, make them appear blue color - we
call such components as basophilic :→ nuclei =
blue
--- Eosin: acidic stain, combines with basic components,
make them appear pink color 🡺 we call such
components as acidophilic (eosinophilic) :→
cytoplasm = pink
Other Stains:
How to use a microscope

https://www.youtube.com/watch?v=SUo2fHZaZCU
High Power Field (X40)
Artifacts
vesiculobullous lesions
Artifacts related to microtomy

Histopathological image shows scaring and tearing

of section due to nick in knife edge (H&E, ×10)
Histopathological image shows wrinkles
and folds due to uneven stretching of
tissue sections (H&E, ×10)
Histopathological image shows air bubbles
(H&E, ×10)
Questions
In preparing tissue for routine light microscopic study, which procedure
immediately precedes clearing the specimen with an
organic solvent ?

A.Dehydration
B.Fixation
C.Staining
D.Clearing
E.Embedding
In preparing tissue for routine light microscopic study, which procedure
immediately precedes clearing the specimen with an
organic solvent ?

A.Dehydration
B.Fixation
C.Staining
D.Clearing
E.Embedding
Which of the following staining procedures relies on the cationic and
anionic properties of the material to be stained?
A.Fixation
B.Dehydration
C.Hematoxylin & eosin staining
D.Mounting
Which of the following staining procedures relies on the cationic and
anionic properties of the material to be stained?
A.Fixation
B.Dehydration
C.Hematoxylin & eosin staining
D.Mounting
In a light microscope used for histology, resolution and magnification
of cells are largely dependent on which component?

A.Condenser
B.Objective lens
C.Eyepieces or ocular lenses
D.Specimen slide
E.The control for illumination intensity
In a light microscope used for histology, resolution and magnification
of cells are largely dependent on which component?

A.Condenser
B.Objective lens
C.Eyepieces or ocular lenses
D.Specimen slide
E.The control for illumination intensity