Abstract
Investigation into the molecular mechanisms regulating normal renal physiology and pathophysiology has benefited from the development of microdissection techniques enabling sampling of specific cell populations or structures within the kidney. Laser-capture microdissection and pressure catapulting is a relatively new, entirely non-contact microdissection technique that facilitates the assay of mRNA and protein expression in single nephron segments or populations. Herein, we describe methods for sample preparation, microdissection and collection of glomeruli from archival renal biopsies for later analysis of gene expression using real-time PCR. Microdissection of glomeruli from archival renal biopsy sections was carried out using the PALM Microbeam UV laser system from P.A.L.M. Technologies.
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References
Kohda, Y., Murakami, H., Moe, O.W., and Star, R.A. (2000) Analysis of segmental renal gene expression by laser capture microdissection. Kidney Int. 57, 321–331.
Nagasawa, Y., Takenaka, M., Matsuoka, Y., Imai, E., and Hori, M. (2000) Quantitation of mRNA expression in glomeruli using laser-manipulated microdissection and laser pressure catapulting. Kidney Int. 57, 717–723.
Schutze, K., Niyaz, Y., Stich, M., and Buchstaller, A. (2007) Noncontact laser microdissection and catapulting for pure sample capture. Methods Cell Biol. 82, 649–673.
Chaudhary, K.W., Barrezueta, N.X., Bauchmann, M.B., Milici, A.J., Beckius, G., Stedman, D.B., Hambor, J.E., Blake, W.L., McNeish, J.D., Bahinski, A., and Cezar, G.G. (2006) Embryonic stem cells in predictive cardiotoxicity: laser capture microscopy enables assay development. Toxicol. Sci. 90, 149–158.
Mayer, A., Stich, M., Brocksch, D., Schutze, K., and Lahr, G. (2002) Going in vivo with laser microdissection. Methods Enzymol. 356, 25–33.
Espina, V., Wulfkuhle, J.D., Calvert, V.S., VanMeter, A., Zhou, W., Coukos, G., Geho, D.H., Petricoin, E.F. 3rd, and Liotta, L.A. (2006) Laser-capture microdissection. Nat. Protoc. 1, 586–603.
Specht, K., Richter, T., Muller, U., Walch, A., Werner, M., and Hofler, H. (2001) Quantitative gene expression analysis in microdissected archival formalin-fixed and paraffin-embedded tumor tissue. Am. J. Pathol. 158, 419–429.
Goldsworthy, S.M., Stockton, P.S., Trempus, C.S., Foley, J.F., and Maronpot, R.R. (1999) Effects of fixation on RNA extraction and amplification from laser capture microdissected tissue. Mol. Carcinog. 25, 86–91.
Burgemeister, R., Gangnus, R., Haar, B., Schutze, K., and Sauer, U. (2003) High quality RNA retrieved from samples obtained by using LMPC (laser microdissection and pressure catapulting) technology. Pathol. Res. Pract. 199, 431–436.
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Edgley, A.J., Gow, R.M., Kelly, D.J. (2010). Laser-Capture Microdissection and Pressure Catapulting for the Analysis of Gene Expression in the Renal Glomerulus. In: Hewitson, T., Darby, I. (eds) Histology Protocols. Methods in Molecular Biology, vol 611. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-345-9_3
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DOI: https://doi.org/10.1007/978-1-60327-345-9_3
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Publisher Name: Humana Press, Totowa, NJ
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