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Requirements for good tissue microarray (TMA) results

Requirements for good tissue microarray (TMA) results. Thick donor tissue blocks At least 2 mm Best if > 3mm Guide slide must represent the last section cut from the block Selection of target tissue most likely to be present through full thickness of the block

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Requirements for good tissue microarray (TMA) results

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  1. Requirements for good tissue microarray (TMA) results • Thick donor tissue blocks • At least 2 mm • Best if > 3mm • Guide slide must represent the last section cut from the block • Selection of target tissue most likely to be present through full thickness of the block • “Tight” circling of targets required Back to home page

  2. Use of histologic sections that are not representative of the cut surface of the block, and imprecise circling leads to the missing of target tissue, due to the “blind” nature of tissue core sampling during the TMA manufacture. Back to home page

  3. Tissue architecture changes with depth of sectioning Tumor with solid and cystic components: histologic sections taken from different levels of the block. Note in particular that the cystic areas and the edges of the solid tumor change in position within the tissue. Back to home page

  4. Section of breast tissue: A heterogeneous population of adipose, fibrous and epithelial structures. Back to home page

  5. The desired target tissue: normal breast epithelium Back to home page

  6. Imprecise circling may lead the maker of the TMA to miss the target tissue. Back to home page

  7. Approximate needle size Back to home page

  8. Tighter circling of the area will result in a higher success rate in capturing target tissue. Back to home page

  9. In tissues with widely-dispersed target tissue, another strategy is to circle small areas for individual needle placement. Back to home page

  10. Back to home page

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  12. Some tissues will have target tissue that contains internal undesirable areas. Back to home page

  13. Tight circling may still result in non-target areas being sampled Back to home page

  14. Fill in these areas to help guide the maker of the TMA. Back to home page

  15. Problems inherent to tissue • Tissue targets smaller than the thickness of the block, and or small targets which do not maintain a vertical orientation will not be present in all sections of a TMA • Breast ducts and lobules are prime examples Back to home page

  16. Problems inherent to tissue:small size of target tissue elements “Some well-differentiated cancers or other smaller lesions, as well as normal glandular tissues, retained their morphology for a few dozen sections” Olli Kallioniemi in Tissue microarrays for high-throughput molecular profiling of tumor specimensNature Medicine 7:844, 1998 Back to home page

  17. The ideal target tissue architecture is a cylinder, which is the full thickness of the paraffin block. Back to home page

  18. This shape would ensure that target tissue is present in the sampled tissue core in every section of the tissue microarray. Back to home page

  19. Unfortunately, many tissue elements change their location in the paraffin block and/or are not full thickness. This cannot be fully predicted from examination of the surface of the block. Back to home page

  20. To address this issue, it is wise to consider having more than one core per target tissue in the TMA, and to “double stack” the cores. This helps to ensure target tissue is present in more sections of the TMA. Back to home page

  21. Approaches to dealing with“difficult” tissue targets • Use more than one core to sample tissue • Use larger core sizes • “Doublestack” cores Back to home page

  22. TMA block, with tissue core diameters as indicated 2.0 mm 1.0 mm 0.6 mm 1.5 mm Back to home page

  23. TMA spot sizes:microscopic images 20 X original magnification Back to home page

  24. Double stacking of TMA cores The cores are stacked, one on top of the other in the recipient TMA block. Two cores are removed from target tissue in the donor block. Back to home page

  25. TMA example • A TMA is to be constructed in 4 copies, with two core positions of target tissue per recipient TMA block, with each core position to be double stacked. • Requires 16 donor cores • Uses approximately a “square” of donor target tissue with 4 mm sides Back to home page

  26. Donor tissue after TMA manufacture 16 cores Back to home page

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