Protocol abstract
Nature Protocols 2, - 1185 - 1196 (2007)
Published online: 10 May 2007 | doi:10.1038/nprot.2007.117
Subject Categories: Imaging | Spectroscopy and structural analysis
Imaging mass spectrometry at cellular length scales
A F Maarten Altelaar1, Stefan L Luxembourg1, Liam A McDonnell1, Sander R Piersma1,2 & Ron M A Heeren1
Abstract
Imaging mass spectrometry (IMS) allows the direct investigation of both the identity and the spatial distribution of the molecular content directly in tissue sections, single cells and many other biological surfaces. In this protocol, we present the steps required to retrieve the molecular information from tissue sections using matrix-enhanced (ME) and metal-assisted (MetA) secondary ion mass spectrometry (SIMS) as well as matrix-assisted laser desorption/ionization (MALDI) IMS. These techniques require specific sample preparation steps directed at optimal signal intensity with minimal redistribution or modification of the sample analytes. After careful sample preparation, different IMS methods offer a unique discovery tool in, for example, the investigation of (i) drug transport and uptake, (ii) biological processing steps and (iii) biomarker distributions. To extract the relevant information from the huge datasets produced by IMS, new bioinformatics approaches have been developed. The duration of the protocol is highly dependent on sample size and technique used, but on average takes approximately 5 h.
- FOM Institute for Atomic and Molecular Physics, Kruislaan 407, 1098 SJ Amsterdam, The Netherlands.
- Present address: Oncoproteomics Laboratory, Free University Medical Centre, De Boelelaan 1117, 1081 HV, Amsterdam, The Netherlands.
Correspondence to: Ron M A Heeren1 e-mail: heeren@amolf.nl
nature-products
A-Z product listing
- 4700 proteomics MALDI TOF/TOF analyzer(Applied Biosystems)
- CCD camera(LaVision)
- Chromatography sprayer(Sigma-Aldrich)
- Conductive glass slides(Delta Technologies)
- Cryomicrotome(Leica Microsystems)
- DHB(Fluka)
