Skip to main content

Laser Capture Microdissection of Archival Kidney Tissue for qRT-PCR

  • Protocol
  • First Online:
Kidney Research

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1397))

Abstract

Whole-organ molecular analysis of the kidney potentially misses important factors involved in the pathogenesis of disease in glomeruli and tubules. Organ wide analysis can however be augmented by using laser capture microdissection (LCM) to isolate morphologically similar cells and nephron structures from a heterogeneous tissue section via direct visualization of the cells. The protocol here provides a practical approach utilizing LCM in combination with RNA isolation techniques for downstream analysis. This technique is readily applicable to study mRNA expression in isolated glomeruli and tubules in both experimental animal models and human kidney biopsy material.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Higgins JP, Wang L, Kambham N et al (2004) Gene expression in the normal adult human kidney assessed by complementary DNA microarray. Mol Biol Cell 15:649–656

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Henger A, Schmid H, Kretzler M (2004) Gene expression analysis of human renal biopsies: recent developments towards molecular diagnosis of kidney disease. Curr Opin Nephrol Hypertens 13:313–318

    Article  CAS  PubMed  Google Scholar 

  3. Darby IA, Hewitson TD (2008) In situ hybridization, Molecular biomethods handbook. Humana, Totowa, NJ

    Book  Google Scholar 

  4. Espina V, Wulfkuhle JD, Calvert VS et al (2006) Laser-capture microdissection. Nat Protoc 1:586–603

    Article  CAS  PubMed  Google Scholar 

  5. Kohda Y, Murakami H, Moe OW et al (2000) Analysis of segmental renal gene expression by laser capture microdissection. Kidney Int 57:321–331

    Article  CAS  PubMed  Google Scholar 

  6. Bojmar L, Karlsson E, Ellegard S et al (2013) The role of microRNA-200 in progression of human colorectal and breast cancer. PLoS One 8:e84815

    Article  PubMed  PubMed Central  Google Scholar 

  7. Tanji N, Ross MD, Cara A et al (2001) Effect of tissue processing on the ability to recover nucleic acid from specific renal tissue compartments by laser capture microdissection. Exp Nephrol 9:229–234

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Tim D. Hewitson .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Hewitson, T.D., Christie, M., Smith, E.R. (2016). Laser Capture Microdissection of Archival Kidney Tissue for qRT-PCR. In: Hewitson, T., Smith, E., Holt, S. (eds) Kidney Research. Methods in Molecular Biology, vol 1397. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3353-2_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3353-2_17

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3351-8

  • Online ISBN: 978-1-4939-3353-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics