HISTOLOGY

Definition
 Histology
 Greek word “histo”- means tissue or web
 Branch of morphological science that deals with the study of
 Many cell types and extracellular components
 Tissues arranged into organs
 Cells – tissues - organ

 Microscopic Anatomy
Definition
 Cytology

 Study of cells
 Cells spontaneously shed off
 Cells physically removed by swabs or brushings
Study of Cells

 Tissue section

 Cytology smear
 Study of Cells
Tissue Section
 Steps
 Fixation- to avoid tissue digestion
 Dehydration- series of increasing concentration of alcohol
 Clearing- removal of alcohol
 Impregnation/Infiltration – melted paraffin at 58°C - 60°C
 Embedding- paper boats, paraffin
 Trimming or cutting –to expose the tissue for sectioning
 Sectioning- microtome
 Staining- for visualization, H& E
 Mounting on microscopic slides
 Labeling
Study of Cells
Tissue Section
 Histological staining
 H and E: universal stain
(Hematoxylin and Eosin)
 Histochemical stain
 Special stains- Mallory stain for nuclei
 Enzymes- Gomori Lead method for Acid Phosphatase
 Immune reactions – fluoroscopy
 Immunohistochemical staining
 Anatomic Pathology Biopsy Applications
Study of Cells
Tissue Section
 Hematoxylin and eosin (H & E)

 Hematoxylin stains nucleus and other acidic structures (RNA-

rich portions) blue

 Eosin stains cytoplasm and collagen pink

Study of Cells
Tissue Section
 H and E stain result

 Nuclei: blue to blue black

 Nucleolus: dark blue
 Cytoplasm: pink
 RBC, eosinophil granules, keratin: bright orange-red
Study of Cells
Tissue Section
 H and E stain result
 Basophil cytoplasm, plasma cells, osteoblast: purplish
 Muscle fibers: deep pink
 Collagen tissue: light pink
 Cartilage: light blue to dark blue
 Calcium, calcified bone: purplish blue
 Tissue section
 Hematoxylin and eosin (H & E)
 Hematoxylin stains nucleus and
other acidic structures (RNA-
rich portions) blue
 Eosin stains cytoplasm and
collagen pink
Study of Cells

 Cytology smear

 Papanicolau stain
 Giemsa or Wright stain
 Cytology smear
 Papanicolau’s stain
 Hematoxylin
 H&E
 Wright’s stain
Terminologies
 Transverse/cross-section and Sagittal/longitudinal section
 Basophilic and eosinophilic
 Nucleus: pyknotic and vesicular
 Solid organs: stroma and parenchyma
 Hollow organs: wall(mural) or layers
Terminologies

 Transverse/cross-section and
longitudinal
Terminologies

 Basophilic and eosinophilic

Terminologies

 Nucleus: pyknotic and vesicular

Terminologies

 Solid organs: stroma and

parenchyma
Terminologies

 Hollow organs: wall or layers

Tissue section cuts
Light Microscopy

 Bright-field microscopy
 - widely used; composed of mechanical and optical parts
 Total Magnification(TM)-
 Resolving power- smallest distance between 2 particles at
w/c they can be seen as separate objects
Fluorescence Microscopy
 Tissue sections are irradiated w/ UV light and the emission is in the visible
portion of the spectrum
 Acridine Orange- example of fluorescent stain
Phase Contrast and Differential
Interference Microscopy
 - uses a lens system that produces visible images from
transparent objects
 For tissue cultures and observing living cells
 DIM- produces an image w/ a more apparent 3D aspect than
phase contrast
 Confocal Microscopy
 Avoids stray light and achieves greater resolution by using:
 (1) small point of high intensity light provided by laser and
 (2) a plate w/ a pinhole aperture in front of image detector
Polarizing Microscopy

 Allows the recognition of structures made of highly

organized molecules
 Birefringence- ability to rotate the direction of vibration of
polarized light. A feature of crystalline substances
Electron Microscopy
- beam of electrons can be deflected by EM fields in a
manner similar to that in glass lenses.
 Transmission electron microscopy- permits resolution
around 3mm allowing up to 400,000 times magnification
 Scanning electron microscopy- permits pseudo 3D images
of surfaces of cells