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  • Rapid method of quantification of tight-junction organization using image analysis.

Rapid method of quantification of tight-junction organization using image analysis.

Cytometry. Part A : the journal of the International Society for Analytical Cytology (2012-12-06)
Christine Terryn, Mehdi Sellami, Caroline Fichel, Marie-Danielle Diebold, Sophie Gangloff, Richard Le Naour, Myriam Polette, Jean-Marie Zahm
ABSTRACT

The spatial organization of proteins in a cell population or in tissues is an important parameter to study the functionality of biological specimens. In this article, we have focused on tight junctions which form network-like features in immunofluorescence microscopy images. Usually, the organization or disorganization of tight junctions is noticed qualitatively. The aim of this article is to present a simple method to quantify the organization level of tight junction network using image analysis with a dedicated macro developed with Image J software. The method has been validated with simulated images displaying regular decrease of network organization. Then, the macro has been applied to immunofluorescence microscopy images of cells in culture and of tissue sections.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Elastase from porcine pancreas, Type I, ≥4.0 units/mg protein
Sigma-Aldrich
Elastase from porcine pancreas, Type IV, Protein 50-90 %, lyophilized powder, ≥4.0 units/mg protein (biuret)
Sigma-Aldrich
Elastase from porcine pancreas, lyophilized powder, suitable for cell culture
Sigma-Aldrich
Elastase from porcine pancreas, Type III, lyophilized powder, Protein 55-85 %, ≥4.0 units/mg protein