Abstract
Matrix-assisted laser desorption ionization (MALDI) imaging is widely used for in situ proteomic mapping and finds multiple applications in pathology. However, low fragmentation yields in MALDI avoid an optimal identification of peptides from tissues. On the other hand, LMD-based microproteomic analyses allow for the identification of hundreds to thousands of proteins from small tissue regions. Herein, we present the combination of MALDI imaging and LMD-based microproteomic approaches for parallel identification. We illustrate the workflow with an application to intratumor heterogeneity studies.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Longuespee R, Casadonte R, Kriegsmann M, Pottier C et al (2016) MALDI mass spectrometry imaging: a cutting-edge tool for fundamental and clinical histopathology. Proteomics Clin Appl 10:701–719
Longuespee R, Alberts D, Pottier C, Smargiasso N et al (2016) A laser microdissection-based workflow for FFPE tissue microproteomics: important considerations for small sample processing. Methods 104:154–162
Longuespee R, Casadonte R, Kriegsmann M, Wandernoth P et al (2017) Proteomic investigation of human cystic echinococcosis in the liver. Mol Biochem Parasitol 211:9–14
Alberts D, Pottier C, Smargiasso N, Baiwir D et al (2017) MALDI imaging-guided microproteomic analyses of heterogeneous breast tumors—a pilot study. Proteomics Clin Appl (in press)
Balluff B, Frese CK, Maier SK, Schone C et al (2015) De novo discovery of phenotypic intratumour heterogeneity using imaging mass spectrometry. J Pathol 235:3–13
Fox CH, Johnson FB, Whiting J, Roller PP (1985) Formaldehyde fixation. J Histochem Cytochem 33:845–853
Maree R, Rollus L, Stevens B, Hoyoux R et al (2016) Collaborative analysis of multi-gigapixel imaging data using cytomine. Bioinformatics 32:1395–1401
Tyanova S, Temu T, Cox J (2016) The MaxQuant computational platform for mass spectrometry-based shotgun proteomics. Nat Protoc 11:2301–2319
Herfs M, Longuespee R, Quick CM, Roncarati P et al (2017) Proteomic signatures reveal a dualistic and clinically relevant classification of anal canal carcinoma. J Pathol 241:522–533
Acknowledgments
The authors would like to acknowledge Lisette Trzpiot and Nancy Rosiere for their efficient technical assistance.
Conflict of Interest
The authors have declared no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
Longuespée, R., Alberts, D., Baiwir, D., Mazzucchelli, G., Smargiasso, N., De Pauw, E. (2017). MALDI Imaging Combined with Laser Microdissection-Based Microproteomics for Protein Identification: Application to Intratumor Heterogeneity Studies. In: Sarwal, M., Sigdel, T. (eds) Tissue Proteomics. Methods in Molecular Biology, vol 1788. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_114
Download citation
DOI: https://doi.org/10.1007/7651_2017_114
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7852-6
Online ISBN: 978-1-4939-7854-0
eBook Packages: Springer Protocols