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Expression microdissection adapted to commercial laser dissection instruments

Nature Protocols volume 6, pages 457–467 (2011)Cite this article

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Abstract

Laser-based microdissection facilitates the isolation of specific cell populations from clinical or animal model tissue specimens for molecular analysis. Expression microdissection (xMD) is a second-generation technology that offers considerable advantages in dissection capabilities; however, until recently the method has not been accessible to investigators. This protocol describes the adaptation of xMD to commonly used laser microdissection instruments and to a commercially available handheld laser device in order to make the technique widely available to the biomedical research community. The method improves dissection speed for many applications by using a targeting probe for cell procurement in place of an operator-based, cell-by-cell selection process. Moreover, xMD can provide improved dissection precision because of the unique characteristics of film activation. The time to complete the protocol is highly dependent on the target cell population and the number of cells needed for subsequent molecular analysis.

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Figure 1: Schematic diagrams of xMD and LCM.
Figure 2: Examples of xMD using commercial microdissection instruments.
Figure 3: Handheld xMD.

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Acknowledgements

This research was supported, in part, by the intramural program of the NIH National Cancer Institute, Center for Cancer Research.

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Author notes

  1. Jeffrey C Hanson and Michael A Tangrea: These authors contributed equally to the work.

Authors and Affiliations

  1. Laser Microdissection Core, Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA

    Jeffrey C Hanson, Jaime Rodriguez-Canales & Michael R Emmert-Buck

  2. Pathogenetics Unit, Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA

    Michael A Tangrea, Skye Kim, Michael D Armani & Michael R Emmert-Buck

  3. Signal Processing and Instrumentation Section, Center for Information Technology, National Institutes of Health, Bethesda, Maryland, USA

    Thomas J Pohida

  4. Laboratory of Medical Biophysics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, USA

    Robert F Bonner

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  1. Jeffrey C Hanson
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  6. Robert F Bonner
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  7. Jaime Rodriguez-Canales
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  8. Michael R Emmert-Buck
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Contributions

R.F.B., T.J.P., M.A.T. and M.R.E.-B. developed expression microdissection (xMD). J.C.H., M.A.T. and M.R.E.-B. designed the experiments. J.C.H., S.K., M.D.A. and M.A.T. conducted the tests. J.C.H., J.R.-C., M.A.T. and M.R.E.-B. analyzed the data. J.C.H., M.A.T., J.R.-C. and M.R.E.-B. wrote the manuscript. J.C.H. and M.A.T. contributed equally to the work.

Corresponding author

Correspondence to Michael R Emmert-Buck.

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Competing interests

M.E.-B., R.B., and T.P. are inventors on NIH patents covering laser capture microdissection and xMD technology and can receive royalty-based payments through the NIH technology transfer program.

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Hanson, J., Tangrea, M., Kim, S. et al. Expression microdissection adapted to commercial laser dissection instruments. Nat Protoc 6, 457–467 (2011). https://doi.org/10.1038/nprot.2010.202

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