Abstract
The PALM Robot MicroBeam laser microdissection system can isolate specified cells from complex tissues section, in a rapid and precise manner. Combined with other methods, PALM may be used for gene expression elucidating the role of specialized cell type in physiological and pathological activity. This chapter describes the application of the PALM MicroBeam system to isolate RNA from cells in a complex tissue for subsequent gene expression analysis. Protocols show the steps from preparation of tissue samples to the final quantitative results. The process is articulated in several steps, each of which requires optimal choices in order to obtain reliable data from a limited number of cells (500–10,000 cells). Furthermore, the notes regarding tissue preparation, microdissection of the interested cells, are also emphasized.
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Murray, G.I. (2007). An overview of laser microdissection technologies. Acta Histochem. 109, 171–6.
Wang. G., Lai, F., Lai, K., et al. (2009). Discrepancy between intrarenal messenger RNA and protein expression of ACE and ACE2 in human diabetic nephropathy. Am J Nephrol. 29, 524–31.
Blakey, G., Laszik, Z. (2004). Laser-assisted microdissection of the kidney: fundamentals and applications. J Mol Histol. 35, 581–7.
Pinzani, P., Orlando, C., Pazzagli, M. (2006). Laser-assisted microdissection for real-time PCR sample preparation. Mol Aspects Med. 27, 140–59.
Schutze, K., Becker, B., Bernsen, M., Bjornsen, T., Broksch, D., Bush, C., Clement-Sengewald, A., et al. Tissue microdissection, laser pressure catapulting. In: Bowtell, D., Sambrook, J. (Eds.), DNA microarrays: A Molecular Cloning Manual. Cold Spring Harbor Laboratory Press, Cold Spring, Harbor, NY, 2003:331–56.
Chan, R., Lai, F., Li, E., et al. (2007). Intrarenal cytokine gene expression in lupus nephritis. Ann Rheum Dis. 66, 886–92.
Chan, R., Lai, F., Li, E., et al. (2004). Expression of chemokine and fibrosing factor messenger RNA in the urinary sediment of patients with lupus nephritis. Arthritis Rheum. 50, 2882–90.
Guide to Performing Relative Quantitation of Gene Expression Using Real-Time Quantitative PCR. Foster City, Applied Biosystems, 2004.
Vincek, V., Nassiri, M., Knowles, J., Nadji, M., Morales, A. (2003). Preservation of tissue RNA in normal saline. Lab Invest. 83, 137–8.
LCM Staining Kit. Foster City, Applied Biosystems, 2008.
Burgemeister, R., Gangnus, R., Haar, B., Schütze, K., Sauer, U. (2003). High quality RNA retrieved from samples obtained by using LMPC (laser microdissection and pressure catapulting) technology. Pathol Res Pract. 199, 431–6.
Cohen, C., Gröne, H., Gröne, E., Nelson, P., Schlöndorff, D., Kretzler, M. (2002). Laser microdissection and gene expression analysis on formaldehyde-fixed archival tissue. Kidney Int. 61, 125–32.
Espina, V., Wulfkuhle, J., Calvert, V., et al. (2006). Laser-capture microdissection. Nat Protoc.1, 586–603.
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Lu, JX., Szeto, CC. (2011). Gene Expression Using the PALM System. In: Murray, G. (eds) Laser Capture Microdissection. Methods in Molecular Biology, vol 755. Humana Press. https://doi.org/10.1007/978-1-61779-163-5_3
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DOI: https://doi.org/10.1007/978-1-61779-163-5_3
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Publisher Name: Humana Press
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Online ISBN: 978-1-61779-163-5
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