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INTRODUCTION
The preparation of permanent, stained sections of animal tissue is an
important technique in modern pathology. It allows normal tissue to be
distinguished from abnormal tissue. It involves the following stages:
1. Selection
6. Blocking-out
2. Fixation
7. Sectioning
3. Dehydration
8. Staining
4. Clearing
9. Mounting
5. Wax impregnation
These procedures are carried out routinely in Histology Laboratories for
research and diagnostic purposes. They are very time consuming and the
operation is, therefore, mostly automated using a tissue processor. With the
aid of an automatic tissue processor the entire process can be completed
within 24 hours; equally there are several points where the process can be
stopped and restarted as time permits.
Time constraints, and Health and Safety considerations, do not allow you to
carry out the entire process, but you will have the opportunity to see a
demonstration of some of the procedures and to stain and mount tissue
sections that have been prepared for you.
e.g. formalin
e.g. 70%, 90% and 100% (absolute) alcohol
e.g. Histoclear or xylene
Microtome
H and E Staining
H and E staining is a routine staining procedure carried out to examine basic
tissue architecture. Haematoxylin is a basic dye that will bind to acidic
structures in the cell such as nucleic acid. In acidic conditions the dye
appears red. Sections are first stained with the haematoxylin dye and then
100% Ethanol
Histoclear
70% Ethanol
5
Cover Slips
Harris Haemotoxylin
Stage Micrometer
Eosin Stain
Per Student
1% Acid Alcohol
Microscope
TECHNICAL NOTES
Histoclear II (HS-202), Harris Haematoxylin (HS-400) and Omnimount (HS110) are available from National Diagnostics.
Formalin Fixed Paraffin Embedded tissue sections are available from
Microtechnical Services Ltd for 0.75/slide. A full list of the available sections
is available at their website http://www.microtechnicalservices.co.uk/
Eosin Stain:
1 g Eosin Y dissolved in 100 mLs of deionised water.
1% Acid Alcohol
2.7 mLs of 36% HCl + 70 mLs of ethanol made up to 100 mLs with deionised
water.
Scotts Tap Water Substitute
20 g of MgSO47H2O + 2 g of NaHCO3 made up to 1 L with dH2O.