Flashcards
Slide prep - embedding to staining
Cards in this deck (12)
- 11. EmbeddingAims to create a wax ‘block’ with tissue in it orientated ready for sectioning
Embedding Centre using machine - 2importance of correct orientationMost tissue blocks, samples cut from the largest area of tissue – samples embedded such that largest area is faced down in the mould
- 3exceptions- tubular structures
- muscle biopsies in both transverse and longitudinal
- epithelial tissue - 42. Microtomy- Thin enough to allow light to pass through (4 um )
- Mounted on glass microscope slide - 5Problem - wrinkles on waxSolution - Wrinkles removed by floating sections in warm waterbath (42⁰C)
- 63. Mounting onto slide from waterbath- Slide held in waterbath vertically, lifted out vertically with section adhered to it
- drained & dried at 37-60 C. - 74. Rehydration by xylene - decreasing alcohol conc - water- Dyes used to stain tissue are aqueous, will not penetrate tissue embedded in wax.
- Section rehydration removes wax and replaces with water (reverses tissue processing) – rapid on thin section - 85. staining- Make cell structure visible
- Show variation in cell and tissue structure
- Indicate the chemical nature of cellular/tissue components - 96. Mounting using 0;17mm coverslip for protection. - requirements- Have refractive index close to glass (of coverslip and slide) and tissue
- Be clear under normal use
- Have no effects on stains used
- Have no inherent fluorescence
- Dry/set quickly - 107.Professional's review and quality reassurance- May recommend additional investigations
- internal or external - 11urgent samples using cryotechniuqes and tissue removed by Mohs surgeryresults on frozen samples within 15 minutes
Preserve enzyme activity for functional assay
Avoid fixatives that interfere with specific type of subsequent staining
Rapid diagnosis of neoplasms (benign vs malignant) - 12consSections 5mm or more
Artefacts associated with incomplete freezing ‘waffling’
Fatty tissue poorly sectioned
Size of blocks limited ~1 cm3
Contamination
Sectioning – time consuming
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- BIOL 282 - Cellular pathology Flashcards on Slide prep - embedding to staining, created by Jumael Zafar on 19/11/2019.
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- Jumael Zafar
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