Lipid mapping in human dystrophic muscle by cluster-time-of-flight secondary ion mass spectrometry imaging

Nora Tahallah,Alain Brunelle,Sabine De La Porte,Olivier Laprévote

doi:10.1194/jlr.m700421-jlr200

Abstract

Human striated muscle samples, from male control and Duchenne muscular dystrophy-affected children, were subjected to cluster-time-of-flight secondary ion mass spectrometry (cluster-ToF-SIMS) imaging using a 25 keV Bi(3)(+) liquid metal ion gun under static SIMS conditions. Spectra and ion density maps, or secondary ion images, were acquired in both positive and negative ion mode over several areas of 500 x 500 microm(2) (image resolution, 256 x 256 pixels). Characteristic distributions of various lipids were observed. Vitamin E and phosphatidylinositols were found to concentrate within the cells, whereas intact phosphocholines accumulated over the most damaged areas of the dystrophic muscles, together with cholesterol and sphingomyelin species. Fatty acyl chain composition varied depending on the region, allowing estimation of the local damage extent.

Highlights

Human striated muscle samples, from male control and Duchenne muscular dystrophy-affected children, were subjected to cluster-time-of-flight secondary ion mass spectrometry imaging using a 25 keV Bi31 liquid metal ion gun under static SIMS conditions
We investigated human striated muscle originating from surgery residues of one control (12 year old) and two Duchenne muscular dystrophy (DMD)-affected (13 and 14 year old) children
The myofibers are clearly observed, they are not regularly sized and only a few zones appear more compact. These two images helped us to choose the areas to be analyzed by mass spectrometry imaging on the DMDaffected muscle sections

Summary

Introduction

From male control and Duchenne muscular dystrophy-affected children, were subjected to cluster-time-of-flight secondary ion mass spectrometry (cluster-ToF-SIMS) imaging using a 25 keV Bi31 liquid metal ion gun under static SIMS conditions. Spectra and ion density maps, or secondary ion images, were acquired in both positive and negative ion mode over several areas of 500 3 500 mm (image resolution, 256 3 256 pixels). Characteristic distributions of various lipids were observed. Vitamin E and phosphatidylinositols were found to concentrate within the cells, whereas intact phosphocholines accumulated over the most damaged areas of the dystrophic muscles, together with cholesterol and sphingomyelin species. Fatty acyl chain composition varied depending on the region, allowing estimation of the local damage extent.—Tahallah, N., A. Lipid mapping in human dystrophic muscle by cluster-time-of-flight secondary ion mass spectrometry imaging.

Methods

Results

Discussion

Conclusion