Authors
Marcelina Cardoso Dos Santos, Cyrille Vézy, Rodolphe Jaffiol,
Title
Membrane-substrate separation distance assessed by normalized total internal reflection fluorescence microscopy
In
SPIE Photonics West
Volume
8949
Pages
89491H
Publisher
SPIE
Year
2014
Indexed by
Abstract
As a consequence of the recent progress in nanoscale technology, more and more sensitive methods are developed to characterize and understand the dynamic of cell membrane adhesion process. In this paper we present a new quantitative method to measure the separation distances between the membrane and the substrate. This technique is based on a normalization of Total Internal Reflection Fluorescence (TIRF) images by usual epi-illumination images. This simple method allows to achieve a nanometric axial resolution, typically 10 nm. We demonstrate the potential of our technique through the study of phospholipids membranes such as Giant Unilamellar Vesicles (GUVs), which are usual biomimetic systems to investigate membrane-substrate interactions.
Affiliations
Offprint