1. College of Education Biology Dept.
Microscopic Slides Preparation for Histopathology Examination By
Saman M. Mohammad-Amin
PhD. Biotechnology
Seminar Hall, Am

2. Microscopic Slide Preparation
Examining cells, fragments of tissues or organs, after their vital processes have been stopped by fixation.
Allows the examination of the structure and main characteristics of cells.

3. Microscopic Slide Preparation, Purposes and Usage
Purpose: preserving cellular structures and the relations between them indefinitely
Used in :
Clinics
Morphopathology
Forensics
Research purposes
Educational purposes

4. Steps for Microscopic Slides Preparation
Recoltation
Fixation
Dehydration
Clearing
Infiltration
6. Embedding 7. Sectioning 8. Staining 9. Mounting 10. Labeling

5. Recoltation may obtained by :
Obtaining the sample to be examined from humans, living animals or corps.
Recoltation may obtained by :
Biopsy
Necropsy (Autopsy)

6. Fixation
A physical or chemical process that stops as soon as possible vital cellular processes, maintaining with minimal alteration the shape, volume, spatial and molecular relation between elements.
Formaldehyde, Glutaraldehyde, Acetic acid, Metyl alcohol and Osmium tetraoxide used as the fixative , each with it’s advantages and disadvantages.

7. Dehydrating (fluid) agent:
Dehydration
To remove fixative and water from the tissue and replace them with dehydrating fluid.
Dehydrating (fluid) agent:
Ethanol
Methanol
Isopropanol
Acetone

8. Clearing
Clearing is the transition step between dehydration and infiltration with the embedding medium.
Clearing fluid:
Toluene and xylene
Benzene
Chloroform
Cedar wood oil
Clove oil

9. Infiltration media (materials):
Is the saturation of tissue cavities and cells by a supporting substance which is generally, the medium in which they are finally embedded.
Infiltration media (materials):
Paraffin wax , Gelatin , Celloidin and Plastics

10. Embedding
Is the process by which tissues are surrounded by a medium such as agar, gelatin, or wax which when solidified will provide sufficient external support during sectioning.

11. Sectioning
Is a real art and needs much skills and practices. It is important to have a properly fixed and embedded block.
This is done with a Microtome. The important thing for proper sectioning is a very sharp knife.

12. Staining
Intends to increase the contrast between cell or tissue components by modifying refraction indexes of the morphologic substrate, using dyes, it done as follows:
A. Deparaffinization
B. Hydration
C. Staining: The most common stain applied for histological study is Hemotoxylin and Eosin.
D. Dehydration
E . Clearing

13. There are two types of mounting media :
Histological sections which need to be examined for any length of time or to be stored, must be mounted under a cover-slip.
There are two types of mounting media :
1. Aqueous media: Used for material which is unstained such, (glycerin).
2. Resinous media :For routine staining such, (Canada balsam).

14. Labeling It will be noted: Celullar type Recoltation source
The fixative
The staining
Date

15. Normal tissues appearances
Prostate
Kidney

16. Breast Histopathology
Normal breast tissue
Cancerous breast tissue

17. Thank You For Attention