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

Practical 4: H&E
Dr 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
Histology video of thin skin

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

Skin

Boston
University

Skin

Boston
University

Skin

Boston
University

Skin

Boston
University

GI Tract

Ileum
Histology video of the ileum:
https://
www.youtube.com/watch?v=M6
zcmsy8c-o

Ileum

Boston University

Peyers patches

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 isnt


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)., Wheaters 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