practical 4 - h&e.pptx
TRANSCRIPT
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(Studies in) Structure and Function of Cells and Tissues – BIOM1004/1904; HCSC1904
Practical 4: H&EDr Louise Dunford
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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
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Skin
http://www.youtube.com/watch?v=-KmfEbKmKFw
Histology video of thin skin
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Skin
© Boston University
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Skin
© Boston University
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Skin
© Boston University
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Skin
© Boston University
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GI Tract
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Ileum
https://www.youtube.com/watch?v=M6zcmsy8c-o
Histology video of the ileum:
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Ileum
Peyer’s patches© Boston University
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Ileum
© Boston University
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Ileum
© Boston University
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Haematoxylin & Eosin (H&E)• The most widely used stain around the world• Known as the “international stain”• A simple histological stain with two colours
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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
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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
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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
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Troubleshooting
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Troubleshooting
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Troubleshooting
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Troubleshooting
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Troubleshooting
Not enough stain in the jar
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Troubleshooting
Dye has precipitated (nb, this isn’t H&E)
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Troubleshooting
Dirt has got between the slide and coverslip
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Troubleshooting
A hair has got between the slide and coverslip
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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
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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