Histology

- (Gr. histo, web or tissue, + logos, study) the study of the tissues of the body and of how these tissues are arranged
to constitute organs.
- Histology is the branch of the anatomical sciences that studies the microscopic structures of tissues
- Histology no longer deals with merely the microscopic structure of the body, it also concerns itself with the
body’s function
- It has direct relationship with other discipline such as cytology, biochemistry, physiology, embryology, gross
anatomy, and pathology
- Tissues are made of cells and extracellular matrix
- The small size of cells and matrix components makes histology dependent on the use of microscopes.

Laboratory techniques for specimen preparation

1. Histotechnique
2. Gross examination
3. Fixation
4. Dehydration and clearing
5. Infiltration and Embedding
6. Sectioning
7. Mounting
8. Staining

Staining
 Three classes are:
– Stains that differentiate between acidic and basic components of the cell
– Specialized stains that differentiate the fibrous components of the extracellular matrix
– Metallic salts that precipitate on tissues, forming metal deposits on them

 Hematoxylin and eosin stain

– Most commonly used stain in histology
– Hematoxylin is a base that preferentially colors the acidic components blue
• Stains nucleic acids - basophilic
– Eosin is an acid that colors the basic components of the cell a pinkish color
• Stains cytoplasmic constituents – acidophilic

Other staining method

 Toluidine blue – stains metachromatic tissue component
 Masson trichrome
• Nuclei – dark blue
• Muscle, keratin, cytoplasm – red
• Mucinogen, collagen – light blue
 Weigert elastic stain – stains elastic fibers blue
 Silver stain – stains reticular fibers black
 Iron hematoxylin – stains striations of muscle and rbc black
 Periodic acid Schiff – stains glycogen and carbohydrate-rich molecules magenta
 Wright and Giemsa stains – used for differentiating blood cells

Light microscopy
• Cross section
• Longitudinal section
• Oblique section

Advanced visualization techniques

1. Histochemistry
A method of staining tissue that provides information concerning the presence and location of intracellular and
extracellular macromolecules

2. Immunocytochemistry
Uses fluoresceinated antibodies and antiantibodies to provide more precise intracellular and extracellular localization of
macromolecules than is possible with histochemistry
3. Autoradiography
A method that uses the incorporation of radioactive isotopes into macromolecules, which are then visualized by the use of
an overlay of film emulsion

Other microscopy techniques

1. Confocal microscopy
2. Electron microscopy
3. Freeze-fracture technique

The Cell
- The basic morphological and functional unit of all living things
- Capable of independent existence under favorable conditions
- 3 Basic Functions of the Cell
1. Transport of Molecules
2. Conversion of Energy
3. Reproduction

- The human body

o composed of approx. 1014 or 100 trillion cells in adults,
o more than 250 cell types.

Basic Structures of Cell:

 Cytoplasm
o Organelles
o Inclusions
o fibrillar elements
 Nucleus

Cell membrane
- A tri-laminar structure that is comprise of two layers of phospholipid molecules
- Protective covering that delimits the cell from its surrounding
- Regulates the movement of molecules
- Provides attachment for cytoskeleton
- Receives and sends out stimuli
- Provides binding sites and receptors for enzymes and other substances
- Allows cell-to-cell recognition
- Forms specialized junctions with the cell membrane of adjacent cells

Cytoplasm
= cytosol + formed elements

Formed elements
 Organelles
 Inclusions
 Fibrillar elements

Cytosol (cytoplasmic matrix)

 Cytosol is viscid, translucent, and colloidal in nature
 It is made up of water (70%), inorganic ions, and organic molecules
 Site of biochemical processes
 Provides a suitable milieu for the organelles in performing their functions

Organelles
 Mitochondria
 Ribosomes
 Endoplasmic reticulum (rough and smooth)
 Golgi complex
 Lysosomes
  Peroxisomes
 Centrosome

Mitochondria
o The “powerhouse” of the cell because they generate most of the energy needed by the cell
o Contains enzymes involved in the Kreb’s tricarboxylic acid cycle

Ribosomes
o Site of protein synthesis
o Two types: free and attached (to rER)
 Free ribosomes – proteins for intracellular processes
 Attached ribosomes – intracellular proteins and proteins that are to be exported by the cell

Endoplasmic reticulum
 Rough
o With ribosomes
o Processes proteins that are produced by the ribosomes

 Smooth
o Site for the synthesis of cholesterol and phospholipids and transport of fatty acids and other lipids

Golgi complex
 The Golgi complex further processes, concentrates, sorts, and packages the proteins that it receives from the rER.
 It releases the proteins into the cytoplasmic matrix in the form of secretory vesicles

Lysosomes
 Contain hydrolytic enzymes released by Golgi complex
 Involved in heterophagy and autophagy.
o Heterophagy – digestion of extracellular particles
o Autophagy – digestion of unneeded or senescent cell organelles

Peroxisomes (microbodies)
 Contain oxidases and catalase
 Involved in many metabolic reactions including the catabolism of fatty acids resulting in the formation of acetyl CoA
and hydrogen peroxide.
 Acetyl CoA is needed in some metabolic processes
 Hydrogen peroxide detoxifies various substances and kills microorganisms
 Catalase degrade excess hydrogen peroxide into water and oxygen

Centrosome (MOTC)
 A dense spherical area near the nucleus and many times surrounded by Golgi complex
 Consists of a pair of minutes, short cylindrical bodies called centrioles that are surrounded by granular structure
called centriolar satellite
 The centrosome is where microtubules are assembled
 The centrioles are the sources of the mitotic spindles that appear during mitosis
 The centriole (basal body) is also the source of the cilia and flagellum

Inclusions
1. Fat droplets
2. Glycogen
3. Zymogen granules
4. Pigment granules
5. Crystals
6. Lipochrome pigment granules
7. Lipofuscin pigments
8. Dust particles

Cytoskeleton
 1. Microfilaments
- Made up of F-actin
- Involved in activities that occur in the cell membrane such as exocytosis and endocytosis
- Associated with organelles
- Mainly or partly responsible for the contraction of myoepithelial cells and muscle cells

2. Intermediate filament
 Keratin – epithelial cells
 Desmin – muscle cells
 Vimentin – fibroblasts and muscle cells
 Neurofilament – nerve cells
 Glial filament – glial cells

3. Microtubule
 Formed in the centrosome
 Play a role in the movement of organelles
 Source of mitotic spindles, cilia, and flagella

Nucleus
 Largest structure inside the cell
 Commonly round or spherical, but may occur in various shapes and may sometimes form lobes
 Consists of:
 Nuclear matrix
 Chromatin
 One or more nucleoli
Covered by a nuclear envelope

nuclear envelope
 Continuous of rER membrane
 Has nuclear pores for the exchange of substances

chromatin
 Entangling threads that forms the chromosome
 The nucleus of all human cells contains 46 chromosome (26 somatic pairs and 1 sex chromosome pair), except
for RBCs, which are anucleate, and the developing gametes, which contain only 23 chromosomes.
 Each chromosome consists of a DNA molecule and associated nucleoproteins.
 4 kinds of DNA bases:
1. Adenine
2. Thymine
3. Cytosine
4. Guanine
 Gene – the segment of the DNA molecule within a chromosome that contains the DNA sequence for the
production of a particular protein or nucleic acid
- Humans have between 30,000 to 40,000

nuclear matrix
 Homogenous substance within the nucleus where the chromatin and nucleoli are embedded.
 Composed of water, proteins, metabolites, and ions

nucleolus
 A spherical, highly basophilic structure that is usually eccentrically located in the nucleus
 The principal function is to synthesize ribosomal subunits
Movement of materials across the cell membrane
For smaller molecules
 Diffusion
 Carrier transport
 Active transport

For bigger substances

 Endocytosis (phagocytosis and pinocytosis)
 Exocytosis