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. //old definition
• the branch of the anatomical sciences that studies the microscopic structures of tissues //macro sa anaphy
• no longer deals with merely the microscopic structure of the body, it also concerns itself with the body’s
function
• //tissues may look haphazardly arranged but they are actually perfectly placed when we study their fxs
• direct relationship w: cytology, biochemistry, physiology, embryology, gross anatomy, & pathology**

Tissues: made up of cells and extracellular matrix

• small size of Cs and ECMs cause of dependence on microscopes

Histotechnique Routine method followed in histopath also

1. Gross examination
• Performed by pathologist. Examine tissue spx.
• properties identified: discoloration, changes in consistency, hardening, etc. weight, dimension
• IMPORTANT: identification of pathologic part of tissue to be examined later

2. Fixation /another term: preservation

• Most commonly used agent: 10% formalin
• Decomposition and bacterial growth prevented
• Spx fixed, preserved, + hardened for chemicals used in subsequent steps later

3. Dehydration and clearing

• Dehydration: To remove fixative and intracellular and extracellular water
• Most commonly used dehydrating agent: alcohol
• Clearing: To remove alcohol and prepare for infiltration
• Most commonly used clearing agent: xylene or xylol

// if we can find an infiltrating agent that can mix with water, dehydration and clearing is not required

4. Infiltration and Embedding

• Fill intra and extracellular spaces w a medium.
• Most commonly used: paraffin wax
• Embedding: tissues placed in paraffin blocks

5. Sectioning
• Tissues cut by microtomes into very thin slices: ~5-10 microns. Ideally 1 layer of cells for light to pass

6. Mounting
•
7. Staining
• Identify, differentiate, and visualize tissue
STAINING:
§ Three major classes are:
§ Stains that differentiate between acidic and basic components of the cell
§ E.g. H&E stain. Nucleus: acidic: violet. Cytoplasm: basic: pink or reddish
§ Specialized stains that differentiate the fibrous components of the extracellular matrix
§ E.g. Azan stain. Collagen fibers: blue. Muscle fibers: red.**
§ Metallic salts that precipitate on tissues, forming metal deposits on them
§ E.g. Silver stain. Reticular fibers: black
//mature red blood cells don't have nuclei

Hematoxylin and Eosin Stain

– Most commonly used stain in histology
– A counter-staining method that will help differentiate nucleus from cytoplasm
Hematoxylin Eosin
– base – acid
– colors the acidic components – colors the basic components
– blue – pinkish
– stains nucleic acid (basophilic) – stains cytoplasmic constituents (acidophilic)

Staining (for quiz daw not for exam pero ewan)

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
///Staining method not the fastest way, not the most specific, but cheapest

Cross section Oblique section Longitudinal section

Most common See layers, how they relate w each other In order to assess the organizations of tissues in those
organs: long tissues such as muscle fibers, bone tissues
• / |

ADVANCED VISUALIZATION TECHNIQUES

1. Histochemistry staining method
A method of staining tissue that provides information concerning the presence and location of intracellular and
extracellular macromolecules
2. Immunocytochemistry antigen-antibody reaction**
Uses labeled (fluorescents) antibodies and/or antiantibodies to provide more precise intracellular and
extracellular localization of macromolecules than is possible with histochemistry
3. Autoradiography use radiation to visualize tissues
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 techniques: confocal microscopy - produce 3d image
electron microscopy - spx electron bombardment. External surface of cells
freeze-fracture technique - spx frozen rapidly, then fractured and broken down. Inside of cells

THE CELL
• basic morphological and functional unit of all living things
• Capable of independent existence under favorable conditions even outside the body

3 BASIC FUNCTIONS of the Cell

- Transport of Molecules
// The reason why we can function as human beings. E.g. circulatory system distributing nutrients etc.
- Conversion of Energy
Most cells can produce own energy. ATP from mitochondria. RBCs cannot
- Reproduction
Ability to replicate, produce more cells.

* The Human Body

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

BASIC STRUCTURE
• Cytoplasm
– Organelles
– Inclusions waste products & useful products produced by organelles
– Fibrillar elements cytoskeleton, maintains shape of cell. Allow mvment of ?
• Nucleus
– Storage site for information

PARTS OF THE CELL

• Cell membrane
o A tri-laminar structure that is comprise of two layers of phospholipid molecules
o Fx: protect cell from extracellular substances (+changes in pH, hormones, enzymes)
o Phospholipid = hydrophobic tail (fatty acid), hydrophilic head (phosphate group and glycerol)
Fxs:
• Protective covering that delimits the cell from its surrounding Divides intracellular and extracellular
• Regulates the movement of molecules even water cannot easily pass through
• Provides attachment for cytoskeleton
• Receives and sends out stimuli e.g. functions of the neurons.
• Provides binding sites and receptors for enzymes and other substances e.g. hormones and antibodies.
• Allows cell-to-cell recognition e.g Activity of the WBCs and neurons**
• Forms specialized junctions with the cell membrane of adjacent cells

• Cytoplasm
o cytosol + formed elements
cytosol: fluid portion formed elements: solid structures
o Cytosol (cytoplasmic matrix)
§ Cytosol is viscid, translucent, and colloidal in nature
§ 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
o Formed elements
§ Organelles
• Mitochondria
• Ribosomes
• Endoplasmic reticulum (rough and smooth)
• Golgi complex
• Lysosomes
• Peroxisomes
• Centrosome
§ Inclusions
• Fat droplets
• Glycogen
• Zymogen granules
• Pigment granules
• Crystals
• Lipochrome pigment granules
• Lipofuscin pigments
• Dust particles
§ Fibrillar elements
ORGANELLES
- 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 – production of proteins for intracellular processes
• Enzymes used by lysosomes, proteins needed by peroxisomes, & components of
cytoskeleton
§ Attached ribosomes – intracellular proteins and proteins that are to be exported by the cell
• Found on surface of rough endoplasmic reticulum (rER)
- Endoplasmic reticulum
o Rough
§ With ribosomes
§ Processes proteins that are produced by the ribosomes
§ Prepare proteins for transport to golgi complex
§ Mainly involved in protein synthesis
o Smooth
§ Site for the synthesis of cholesterol and phospholipids and transport of fatty acids and other lipids
§ Mainly involved in lipid synthesis

- Golgi body / golgi complex

o further processes, concentrates, sorts, and packages the proteins that it receives from the rER.
o releases the proteins into the cytoplasmic matrix in the form of secretory vesicles:
§ Like cell membrane. Composed of phospholipids. Will merge with location’s membrane
o Transportation unit of the cell. Transports products produced by the cell
 - Lysosomes
o Like soldiers inside cells. Lysosomes releases enzymes that destroys the microorganisms na nakuha ng
WBCs
o Contain hydrolytic enzymes released by Golgi complex //they denature proteins, but not specific
o Involved in heterophagy and autophagy.
§ Heterophagy – digestion of extracellular particles e.g. microorganisms ingested by wbc
§ Autophagy – digestion of unneeded or senescent cell organelles e.g. dead organelles
o // Activity of lysosomes are inhibited by FIXATIVES

- Peroxisomes (microbodies)
o Contain oxidases and catalase
o Involved in many metabolic reactions including the catabolism of fatty acids resulting in the formation of
acetyl CoA and hydrogen peroxide.
o Acetyl CoA is needed in some metabolic processes
o Hydrogen peroxide detoxifies various substances and kills microorganisms
§ Potent substance can destroy cell wall of bacteria
o Catalase degrade excess hydrogen peroxide into water and oxygen
o // involved in both production and limiting the activity of hydrogen peroxide

- Centrosome (MOTC)
o A dense spherical area near the nucleus and many times surrounded by Golgi complex
o Consists of a pair of minute, short cylindrical bodies called centrioles that are surrounded by granular
structure called centriolar satellite
o The centrosome is where microtubules are assembled
o One of the main fx: during mitosis
§ The centrioles are the sources of the mitotic spindles that appear during mitosis
o The centriole (basal body) is also the source of the cilia and flagellum
§ Near nucleus: centriole, on other cells (cilia, flagellum): basal body

INCLUSIONS
Fat droplets Storage form of lipids found in adipoce tissues
Glycogen Storage form of glucose found mostly in liver cells
Zymogen granules Storage form of enzymes
Pigment granules Responsible for giving color. (eg melanin)
crystals Unknown function or purpose. Not even mentioned if pathologic or normal.
Found in cells of testes such as Sertoli and Leydig cells. Also in macrophages**
Lipochrome pigment granules
Lipid-based color pigments**
Lipofuscin pigments
Dust particles Not normal inclusions. // More in number than actual lung cells
Found in cytoplasm of macrophages in our lungs

• Cytoskeleton
o 3 types: microfilaments, intermediate filaments, microtubules
Microfilaments Intermediate filaments microtubules
Made up of F-actin Keratin – epithelial cells Largest cytoskeleton
Involved in exocytosis & endocytosis Desmin – muscle cells Formed in the centrosome
Associated with organelles /abundant in smooth muscle fibers Play a role in the movement of
Vimentin – fibroblasts & muscle cells organelles
 Mainly or partly responsible for the Neurofilament – nerve cells Source of mitotic spindles, cilia, and
contraction of myoepithelial cells* Glial filament – glial cells flagella
and muscle cells

• Nucleus
o Largest structure inside the cell
o Commonly round or spherical, but may occur in various shapes and may sometimes form lobes
o Consists of:
§ Nuclear matrix
§ Chromatin
§ One or more nucleoli
o Covered by a nuclear envelope
• Nuclear envelope
o Continuous with rER membrane
o Has nuclear pores for the exchange of substances
• Chromatin
o Entangling threads that forms the chromosome
o The nucleus of all human cells contains 46 chromosome (22 somatic pairs and 1 sex chromosome pair),
except for RBCs, which are anucleate, and the developing gametes, which contain only 23 chromosomes.
o Each chromosome consists of a DNA molecule and associated nucleoproteins.
o 4 kinds of DNA bases:
§ Adenine, Thymine, Cytosine, 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
• Diff. concentration of substances in the body
• Nuclear matrix
o Homogenous substance within the nucleus where the chromatin and nucleoli are embedded.
o Composed of water, proteins, metabolites, and ions
• Nucleolus
o A spherical, highly basophilic structure that is usually eccentrically located in the nucleus
o principal fx: synthesize ribosomal subunits

MOVEMENT OF MATERIALS ACROSS THE CELL MEMBRANE:

For smaller molecules
1. Diffusion - Movement of solutes across the cell membrane governed by the concentration gradient
2. Carrier transport - Also diffusion. Just requires carriers. Carriers can be saturated
3. Active transport - Requires energy. Kahit iba conc. gradient.

For bigger substances // Vit. B. iodine, etc.

1. Endocytosis (phagocytosis and pinocytosis)
2. Exocytosis - Release of substances from the cell