CYTOLOGY

1
CYTOLOGY
CYTOLOGY
BY Dr. TAREK ATIA Histology and Cell Biology
2
BOOK

• Color Textbook of
• Histology
• Leslie P Gartner
• James L Hiatt

3

• - Micro-techniques for tissue preparation.
• - Stains.
• - Microscopes.
• - Special techniques used for medical diagnosis.
• - Cell, Tissues and Organs.

4
Micro-techniques for tissue preparation

• - Using Light or Electron Microscope.
• 1- Fixation-
• - Buffered formol saline
• - 10 formalin
• - Suza, Bouin, Zenker solution
• - Formaldehyde or Glutradhyde
• - Osmium tetraoxide
• - Potassium permanganate

5

• 2- Dehydration Gradual removal of water
  from the tissue using ascending grads of
  ethyl alcohol to prevent tissue shrinking.
• 3- Clearing Replacement of alcohol in tissue by
  clearing fluid like xylen, benzene, or acetone

6

• 4- Embedding
• - Tissues are impregnated in paraffin
• - Tissues are impregnated in Epon in
  gelatin capsule.
• 5- Cutting
• - Paraffin block are cut by microtome
  using metal knife, into thin
  sections 6µ
• - Capsules are cut by ultra-microtome,
  using glass or diamond knife,
  into ultra thin sections
  50-100nm.

7

• 6- Mounting
• - Sections spread on the hot plate and
  mounted on glass slides.
• - Sections mounted on metal grids.
• 7- Staining
• - Variable stains are used for specific
  tissues.
• - Stained by heavy metals like lead
  nitrate and uranyl acetate.

8
(No Transcript)
9
(No Transcript)
10
Stains

• - Special dyes used to stain the histological
  sections and make them ready for
  microscope examination.
• - Heamatoxylin and Eosin are most common
  used.

11
Types of stains

• - Reaction of stain
• - Acidic Eosin stain
• - Basic Heamatoxylin
  stain
• - Neutral Leishman stain
• - Physical stain stain dissolve in tissue
  without any chemical reaction such as
  - SUDAN III for fatty tissues.

12

• - Vital stain Staining living tissue inside the
  body. Trypan blue stain.
• - Supravital stain Staining living tissues
  outside the body. Brilliant cresyl
  blue.
• - Metachromatic stain Staining the tissues with
  a color different from the original color of
  stain. Toluidin blue staining of Mast
  cells.

13

• - Polychromatic stain Staining the tissues with
  multiple colors in spite of using a single
  stain. Geimsa stain for blood.
• - Orthocromatic stain Staining the tissues with
  the same color of the stain, such as HE.
• - Histo-chemical stain Staining the different
  chemical components of the cell.
• - Immuno-histo-chemical stain Localisation and
  staining specific proteins by the antigen
  antibody reaction.

14
MICROSCOPY

• - Light microscope
• - Phase contrast microscope
• - Polarizing microscope
• - Fluorescence microscope

• - Electron microscope Scanning / Transmission
• - Interference microscope
• - Confocal microscope
• - Inverted microscope

15
Light microscope
16
(No Transcript)
17
Light microscope
Electron microscope
18
(No Transcript)
19
Special techniques used for diagnosis

• - Histchemistry
• - Immunohistochemistry
• - Tissue / Cell Culture
• - Cell Fractionation
• - Exfoliative Cytology
• - Cytogenetics and Molecular Cytogenetics
• - Bone Marrow smear / Blood film
• - Fine needle aspiration / biopsy
• - Flowcytometry - Autoradiography

20
Cell fractionation
21
Histology and Cytology
22
Course of Histology
- Cytology Cell, Nucleus, DNA, and
Cytogenetics. - Tissues 4 basic tissues
including Epithelium,
Connective (Cartilage, Bone, Blood),
Muscular, and Nervous tissues. - Body
systems and different Organs.