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A Simple and Rapid Protocol to Non-enzymatically Dissociate Fresh Human Tissues for the Analysis of Infiltrating Lymphocytes
Published on: December 6, 2014
A simple, rapid, and convenient Luminex-compatible method of tissue isolation
Derek Barclay1, Ruben Zamora, Andres Torres
1Department of Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania 15213, USA.
Journal of Clinical Laboratory Analysis
|July 16, 2008
Summary
A new method allows for the analysis of protein biomarkers in tissues using Luminex technology. This technique, using RNAlater, offers a simple and rapid approach for tissue biomarker assessment in various clinical settings.
Area of Science:
- Biochemistry
- Molecular Biology
- Clinical Chemistry
Background:
- Pathological conditions often involve changes in multiple protein biomarkers.
- The Luminex platform with xMAP beads is a common tool for biomarker analysis, typically in bodily fluids.
- Measuring biomarkers directly in tissue samples (e.g., biopsies) is often clinically desirable but challenging.
Purpose of the Study:
- To develop a simple and rapid method for tissue biomarker analysis using the Luminex platform.
- To enable the measurement of inflammatory biomarkers (cytokines) directly from excised tissues.
Main Methods:
- Developed a tissue isolation protocol involving overnight permeabilization of excised tissues in RNAlater at 4°C.
- Utilized the Luminex platform for subsequent analysis of inflammatory biomarkers (cytokines).
- Compared the novel RNAlater method with traditional flash-freezing techniques.
Main Results:
- The RNAlater method proved to be a simple and rapid technique for tissue biomarker isolation.
- Successful analysis of inflammatory biomarkers was achieved from both mouse liver and human debrided wound specimens.
- The method demonstrated comparable utility to flash-freezing for tissue biomarker assessment.
Conclusions:
- A novel, efficient method for tissue biomarker analysis using Luminex has been established.
- This RNAlater-based approach facilitates the study of cytokines and other protein biomarkers in tissue samples.
- The technique holds potential utility for various clinical applications, including the analysis of biopsy specimens.
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