Introduction to Histology

Bismark Oliver C. Lemana, M.Sc.

Biological Sciences Department
College of Science, UST

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 Histology as a Science
• G: histo – web or tissue
• Science of tissues (cells + ECM)
– Arrangement to constitute an organ
• Fundamental Types
– Epithelial
– Connective
– Muscular
– Nervous

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 Tissue Composition
• Cells + extracellular matrix

Small intestine x.s.

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 Histological Sample Preparation
50% 75% 100%

• Fixation
– 37% formaldehyde/glutaraldehyde
– Osmium tetroxide (EM)
• Dehydration
– Alcohol series

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 Histological Sample Preparation

• Clearing
– Xylene/xylol
• Infiltration
– paraffin
• Embedding
– Paraffin or resin

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 Sectioning (Microtome)

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 Sectioning (Cryostat)
• Freezing microtome

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 Staining
• Tissues are colorless after dehydration
• Distinction between different tissues
• Basic dyes – basophilic (acidic
structures or molecules)
– Hematoxylin, azures, cresyl violet
• Acidic dyes – acidophilic structures
– Eosin, PAS, Picro-sirius

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 Hematoxylin-Eosin (H&E)
• Purplish blue, blue-black (H); pinkish
red (E)

Section of the stomach

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 Pararosaniline-toluidine blue (PT)
• Purple (chromatin); light violet
(cytoplasm, collagen)

Parotid Gland: (a) PT; (b) H&E

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 Mallory Trichrome
• Purple (nucleus); bright red
(cytoplasm, keratin, erythrocytes);
blue (collagen)

Fibrocartilage: (a) Mallory Trichrome; (b) H&E

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 Picro-sirius-hematoxylin (PSH)
• Collagen (red); cytoplasm (light
violet/pink

Section of kidney

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 Wright-Giemsa Stain
• Differential for leukocytes;
erythrocytes (pink-orange)

Blood cells

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 Silver or Gold Stain
• Neurons/reticulin fibers (dark
brown/black)

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 Stain for Lipids
• Oil red O; Sudan black
• Osmium tetroxide (black)

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 Other stains
• Azures, cresyl violet, brilliant cresyl
blue, luxol fast blue, light green
• Basic aniline dyes
• Cell and extracellular structures
• Paraffin sections
• Bright permanent colors

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 Microscopy
• Light microscopy
– Bright-field
– Fluorescence
– Phase-contrast
– Differential interference
– Confocal
– Polarizing
• Electron microscope
– SEM & TEM

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 Bright-Field Microscope

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 Flourescence Microscopy

Acridine orange DAPI (4’,6-diamino-2-

phenylindole)
Kidney cells

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 FLUORESCENCE MICROSCOPY
A fluorophore (or fluorochrome, similarly to a chromophore) is a fluorescent
chemical compound that can re-emit light upon light excitation.

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 D5 OIM -7M –ru486 D5 OIM -7M –ru486

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 Unstained cells

Bright field microscope Phase-contrast microscope Differential Interference

microscope

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 Confocal Microscope

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 Polarizing microscope

Bright-field Polarizing

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 Transmission Electron microscopy

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 Scanning electron micrscope

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 Cell and Tissue Culture
• In vitro culture of cells or tissues
• Advantages:
– Cultured cells can be obtained in large
quantities
– Most cultures contain only one type of
cell
– Ease in observing cellular activities
– Ability of cells to differentiate
– Cell in vitro mimic their activity in vivo

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 Types of Cell Cultures
• Primary cell cultures
– From primary tissue explants
– Finite life span in culture
– Tissues dissociated into single cells
– Impure cultures
• Continuous cell lines
– Abnormal or transformed cells
– Viral or chemical treatment to
immortalize cells
– Indefinite life span (?)

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 D5 OIM -7M –ru486 D5 OIM -7M –ru486

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 Histo- and Cytochemistry
• Localizing cellular structures in tissue
sections
• utilizes enzymatic activities in
structures
1. Tissue immersion substrate solution
2. Enzyme of tissue acts on substrate
3. Marker compound gets in contact with
tissue
4. Compound reacts with the product of
enzyme-substrate reaction
5. Precipitation of enzyme site

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 Histo- and Cytochemistry

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 Three-dimensional Tissues

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 Short Quiz 1
Identify the type of Specific microscope
used on the following specimen

1.

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 2.

3.

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 4-7. Identify the 4 basic types of
tissues
TRUE or FALSE
8. Hematoxylin can be used as
differential stain for PT
9. The first step in tissue preparation is
infiltration
10. Cryostat is used as a sectioning tool
followed by embedding.

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