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An Alternative Technology to Prepare Tissue Microarray Using Frozen Tissue Samples

  • Zhongting HuEmail author
  • Elbert Chang
  • Melissa Hodeib
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 664)

Abstract

Although most tissue microarray (TMA) slides are currently made from paraffin-embedded tissues, ­frozen clinical tissues are also gradually being used to prepare TMAs. This is because frozen tissues contain better quality RNAs and proteins for profiling gene expressions. Here, we introduce another TMA method that is applicable to a broader range of frozen tissue samples.

In this method, an agarose-gel-based array recipient block is first made using several simple instruments. Frozen donor tissues are then manually cored and arrayed into the recipient block array at −10°C. After arraying, the array block can be immediately sectioned on a cryostat microtome to make TMA slides for in situ hybridization and immunocytochemistry studies. TMAs made by this method have well-defined array configurations, good tissue/cell morphology, and well-preserved proteins and mRNAs. This low-cost and time-saving method provides an alternative tool for preparing high quality TMAs for gene expression analyses.

Key words

Recipient block Frozen tissues Coring Tissue arraying mRNAs Proteins 

Notes

Acknowledgments

We would like to thank Dr. Amy Chew for comments. This research is supported by NIH grants 1R43GM074318-01 and 1R44CA119651-01, to Z. Hu.

References

  1. 1.
    Kononen, J., L. Bubendorf, A. Kallioniemi, M. Barlund, P. Schraml, S. Leighton, J. Torhorst, M.J. Mihatsch, et al. (1998). Tissue microarrays for high-throughput molecular profiling of tumor. Nat Med 4:844–847.PubMedCrossRefGoogle Scholar
  2. 2.
    Scicchitano, M.S., D.A. Dalmas, M.A. Bertiaux, S.M. Anderson, L.R. Turner, R.A. Thomas, R. Mirable, R.W. Boyce (2006). Preliminary comparison of quantity, quality, and microarray performance of RNA extracted from formalin-fixed, paraffin-embedded, and unfixed frozen tissue samples. J Histochem Cytochem 54:1229–1237.PubMedCrossRefGoogle Scholar
  3. 3.
    Hoos, A., C. Cordon-Cardo (2001). Tissue microarray profiling of cancer specimens and cell lines: opportunities and limitations. Lab Invest 81:1331–1338.PubMedCrossRefGoogle Scholar
  4. 4.
    Fejzo, M.S., D.J. Slamon (2001). Frozen tumor tissue microarray technology for analysis of tumor RNA, DNA, and proteins. Am J Pathol 159:1645–1650.CrossRefGoogle Scholar
  5. 5.
    Zhou, L., Hodeib, M., Mendoza, L., Abad, J.D., Z. Hu (2006). New tissue microarray technology for analyses of gene expression in frozen pathological samples. Biotechniques 43:101–104.CrossRefGoogle Scholar
  6. 6.
    Simon, R., M. Mirlacher, G. Sauter (2004). Tissue microarrays. Biotechniques 36:98–105.PubMedGoogle Scholar
  7. 7.
    Waschek, J.A., R.A. Casillas, T.B. Nguyen, E.M. DiCicco-Bloom, E.M. Carpenter, W.I. Rodriguez (1998). Neural tube expression of pituitary adenylate cyclase-activating peptide (PACAP) and receptor: potential role in patterning and neurogenesis. Proc Natl Acad Sci 95:9602–9607.PubMedCrossRefGoogle Scholar
  8. 8.
    Fernebro, E., M. Dictor, P.O. Bendahl, M. Ferno, M. Nilbert (2002). Evaluation of the tissue microarray technique for immunohistochemical analysis in rectal cancer. Arch Pathol Lab Med 126:702–705.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of Anatomy, College of Osteopathic Medicine of the PacificWestern University of Health SciencesPomonaUSA
  2. 2.Research and DevelopmentNeuBiogene Inc.PasadenaUSA

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