1. Professor KI-JONG RHEE Teaching assistant MIN HO LEE

2. Introduction  Diagnostic Histology Cell Biology, Histology, Pathology and Immunology Widely used in diagnostic medicine and bioscience Applied to researches and diagnosis Department of Pathology in hospitals

3. Synopsis  Sampling tissue  Fixation  Washing  Dehydration  Clearing  Infiltration  Embedding  Sectioning  Staining  Mounting

4. Sampling/Fixation/Washing  Dissection of mice Harvest organs liver, spleen, kidney, small intestine, heart, thyroid  Fixation Prevent tissues from being structurally changed Help tissues to be easily stained 10% Formalin, Bouin, D.B., B-5, Zenker, Helly, Carnoy, Orth, Kaiserling  Washing Flowing tap water Remove fixative

5. Dehydration/Clearing/Infiltration  Dehydration Water  Alcohol  Clearing Alcohol  Xylene  Infiltration Xylene  Paraffin Hardening before microsectioning Automatic tissue processor

6. Embedding/Sectioning  Embedding Making a tissue blocks for sectioning  Sectioning Making thin tissue slices 6um

7. Staining/Mounting  Staining determine specific cells, tissues or components based on colour and light refraction Routine staining : Hematoxylin & Eosin stain Special staining : PAS, van Gieson, Verhoeff, Alcian blue, Osmium tetroxide…  Mounting Sealing preparats in the mounting media and a coverglass Prevents physical damage, colour loss and decomposition Water soluble (eg. Glycerol jelly) and insoluble (eg.Canada balsam) mounting media

8. Schedule  9/1Introduction  9/8Tissue preparation (fixation, washing, dehydration, clearing, infiltration)  9/15Embedding  9/22Sectioning  9/29Sectioning  10/6Staining(I)  10/13Staining(II)  10/20Mid-term exam  10/27Preparation of tissues for cryosections  11/3Cryosectioning  11/10Immunohistochemistry (I)  11/17Immunohistochemistry (II)  11/24Special staining (I)  12/1Special staining (II)  12/8Overview  12/15final exam

9. Lab Schedule  Thursday  ClassA4 pm ClassB6 pm  118 Mi-Rae hall (laboratory)

10. Score  Attendence20  Final test30  Report ? Quiz?50

11. Pre-Report  any kinds of normal notebook  Concise  Untill monday morning  Anatomic chart of the mouse Order of dissection Which organ is where  Fixation and washing Kinds of fixatives and purpose for each Why do you fix and wash tissues What is the problem if the tissues are not properly fixed or washed

12. Safety precautions  Wear a lab gown in the laboratory  Beware of inflammable reagents  Liquid paraffin is very hot  Scissors and blades are very sharp  Maintain a quiet atmosphere