Immunofluorescence laser micro-dissection of specific nephron segments in the mouse kidney allows targeted downstream proteomic analysis

Radmila Micanovic, Shehnaz Khan, Tarek Ashkar (El-Achkar)

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Laser micro-dissection (LMD) is a very useful tool that allows the isolation of finite areas from tissue specimens for downstream analysis of RNA and protein. Although LMD has been adapted for use in kidney tissue, the use of this powerful tool has been limited by the diminished ability to identify specific tubular segments in the kidney. In this study, we describe a major improvement in the methodology to isolate specific cells in the mouse kidney using immunofluorescence LMD (IF-LMD). Using IF-LMD, we can reproducibly isolate not only glomeruli, but also S1-S2 proximal segments, S3 tubules, and thick ascending limbs. We also demonstrate the utility of a novel rapid immunofluorescence staining technique, and provide downstream applications for IF-LMD such as real-time PCR and cutting-edge proteomic studies. This technical breakthrough may become an invaluable tool for understanding cellular and molecular events in the heterogeneous kidney milieu.

Original languageEnglish (US)
Article numbere12306
JournalPhysiological Reports
Volume3
Issue number2
DOIs
StatePublished - 2015

Fingerprint

Nephrons
Proteomics
Fluorescent Antibody Technique
Dissection
Lasers
Kidney
Real-Time Polymerase Chain Reaction
Extremities
RNA
Staining and Labeling
Proteins

Keywords

  • 2D-DIGE
  • Laser micro-dissection
  • Mass spectrometry
  • Proteomics

ASJC Scopus subject areas

  • Physiology (medical)
  • Physiology

Cite this

Immunofluorescence laser micro-dissection of specific nephron segments in the mouse kidney allows targeted downstream proteomic analysis. / Micanovic, Radmila; Khan, Shehnaz; Ashkar (El-Achkar), Tarek.

In: Physiological Reports, Vol. 3, No. 2, e12306, 2015.

Research output: Contribution to journalArticle

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