GE Healthcare decyder software 2D DIGE gels were scanned directly after completion of electrophoresis with a Typhoon 9400 scanner (GE Healthcare). 2D DIGE Workflow: An Applied Quantitative Proteomics tool to identify molecular signatures for uro-genital dysfunction in diabetic rat model. Aliens Extermination Rom. Elizabeth Elizabeth.

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. • 2D‐DIGE • laser micro‐dissection • mass spectrometry • proteomics. Introduction Laser micro‐dissection (LMD) is a tool developed in the last two decades that allows direct visualization of tissue sections and capture/isolation of a finite number of cells (Espina et al.

These cells can be used for downstream applications such as RNA or protein extraction and subsequent analyses. In the kidney, LMD was first described by Star and colleagues, who were successful in isolating various tubular segments using rapid immunohistochemical techniques (Kohda et al. Subsequently, the same group adapted immunofluorescence to LMD (IF‐LMD), and were able to isolate thick ascending limbs (TAL) using a specific marker (Murakami et al.

Since those seminal studies, the use of LMD in kidney tissue has been mostly focused on isolation of glomeruli, which are easily recognizable (Pietrzyk et al.; Woroniecki and Bottinger; Wang et al.; Sethi et al. It is not fully clear why the use of LMD to isolate various tubular segments has not gained traction, despite important advances (Noppert et al.; Wilkinson et al. The cause could be partly due to the difficulty in recognizing tubular segments, especially to nonexpert users. With the availability of newer, user‐friendly platforms for LMD, and the access to novel reagents for tissue handling and processing, the ability to reliably dissect specific tubular segments for RNA and protein isolation is now reachable.