(Studies in) Structure and Function of Cells and Tissues – BIOM1004/1904; HCSC1904

Practical 4: H&EDr Louise Dunford

Learning outcomes

• Identify skin & ileum sections

• Describe the different regions/structures within your stained section

• Review slides for quality of staining

• Understand how staining can be improved

Skin

http://www.youtube.com/watch?v=-KmfEbKmKFw

Histology video of thin skin

Skin

© Boston University

Skin

© Boston University

Skin

© Boston University

Skin

© Boston University

GI Tract

Ileum

https://www.youtube.com/watch?v=M6zcmsy8c-o

Histology video of the ileum:

Ileum

Peyer’s patches© Boston University

Ileum

© Boston University

Ileum

© Boston University

Haematoxylin & Eosin (H&E)• The most widely used stain around the world• Known as the “international stain”• A simple histological stain with two colours

Haematoxylin

• Most staining is a simple nuclear stain and a counterstain

• In most staining protocols the lowest molecular weight dyes are applied first (e.g. haematoxylin), differentiated, and then larger counterstains are applied

• Haematoxylin is a mordant (metal complexing) dye; the cationic/basic dye-metal complex binds to nucleic acids in DNA

Eosin

• Anionic/acid counterstain

• Eosin Y binds to amino acids and most cellular components, but not for example, larger molecular structures such as collagen which are also anionic

• Stains pink/red

Troubleshooting

All stages need to be carried out in optimal conditions:

• Collection of the tissue

• Fixation

• Processing

• Cutting

These were done for you prior to the practical

Troubleshooting

Troubleshooting

Troubleshooting

Troubleshooting

Troubleshooting

Not enough stain in the jar

Troubleshooting

Dye has precipitated (nb, this isn’t H&E)

Troubleshooting

Dirt has got between the slide and coverslip

Troubleshooting

A hair has got between the slide and coverslip

Resources

University of Leeds Histology website:http://www.histology.leeds.ac.uk/skin/index.php

Young, B et al., (2006)., Wheater’s Functional Histology: A Text and Colour Atlas. Churchill Livingstone Elsevier

A troubleshooting guide:http://www.leica-microsystems.com/uploads/media/101_steps_to_better_histology.pdf

Learning outcomes

• Identify skin & ileum sections

• Describe the different regions/structures within your stained section

• Review slides for quality of staining

• Understand how staining can be improved