Journal
PROTEOMICS CLINICAL APPLICATIONS
Volume 13, Issue 1, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/prca.201800029
Keywords
formalin-fixed paraffin embedded tissue; MALDI; reproducibility; tissue typing; workflow
Funding
- BMBF grant as part of the Leading-Edge Cluster Ci3 (Cluster for Individualized Immune Intervention) [FKZ 131A029F]
- German Federal Ministry of Education and Research (KMU-innovativ: Medizintechnik Programme) [13GW0081B]
- NCT Gewebebank Heidelberg
- Eurostars-2 Programme
- German Aerospace Center
- BMBF [13FH8I03IA]
Ask authors/readers for more resources
Purpose To facilitate the transition of MALDI-MS Imaging (MALDI-MSI) from basic science to clinical application, it is necessary to analyze formalin-fixed paraffin-embedded (FFPE) tissues. The aim is to improve in situ tryptic digestion for MALDI-MSI of FFPE samples and determine if similar results would be reproducible if obtained from different sites. Experimental Design FFPE tissues (mouse intestine, human ovarian teratoma, tissue microarray of tumor entities sampled from three different sites) are prepared for MALDI-MSI. Samples are coated with trypsin using an automated sprayer then incubated using deliquescence to maintain a stable humid environment. After digestion, samples are sprayed with CHCA using the same spraying device and analyzed with a rapifleX MALDI Tissuetyper at 50 mu m spatial resolution. Data are analyzed using flexImaging, SCiLS, and R. Results Trypsin application and digestion are identified as sources of variation and loss of spatial resolution in the MALDI-MSI of FFPE samples. Using the described workflow, it is possible to discriminate discrete histological features in different tissues and enabled different sites to generate images of similar quality when assessed by spatial segmentation and PCA. Conclusions and Clinical Relevance: Spatial resolution and site-to-site reproducibility can be maintained by adhering to a standardized MALDI-MSI workflow.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available