Philipp Poeller

Projects: SMART-NAFLD

Institutions: Deutsches Krebsforschungszentrum

https://orcid.org/0000-0002-3924-5756
Josch Pauling

Projects: LiSyM Pillar I: Early Metabolic Injury (LiSyM-EMI), LiSyM network, LiSyM-Krebs Partnering

Institutions: Humboldt-Universität zu Berlin - Institute for Theoretical Biology (ITB), Technical University Munich - Klinikum rechts der Isar

Shotgun Lipidomics Combined with Laser Capture Microdissection: A Tool To Analyze Histological Zones in Cryosections of Tissues.
LiSyM Pillar I: Early Metabolic Injury (LiSyM-EMI)

Abstract (Expand)

Shotgun analysis provides a quantitative snapshot of the lipidome composition of cells, tissues, or model organisms; however, it does not elucidate the spatial distribution of lipids. Here we demonstrate … that shotgun analysis could quantify low-picomole amounts of lipids isolated by laser capture microdissection (LCM) of hundred micrometer-sized histological zones visualized at the cryosections of tissues. We identified metabolically distinct periportal (pp) and pericentral (pc) zones by immunostaining of 20 mum thick cryosections of a healthy mouse liver. LCM was used to ablate, catapult, and collect the tissue material from 10 to 20 individual zones covering a total area of 0.3-0.5 mm(2) and containing ca. 500 cells. Top-down shotgun profiling relying upon computational stitching of 61 targeted selective ion monitoring ( t-SIM) spectra quantified more than 200 lipid species from 17 lipid classes including glycero- and glycerophospholipids, sphingolipids, cholesterol esters, and cholesterol. Shotgun LCM revealed the overall commonality of the full lipidome composition of pp and pc zones along with significant ( p < 0.001) difference in the relative abundance of 13 lipid species. Follow-up proteomics analyses of pellets recovered from an aqueous phase saved after the lipid extraction identified 13 known and 7 new protein markers exclusively present in pp or in pc zones and independently validated the specificity of their visualization, isolation, and histological assignment.

Authors: O. Knittelfelder, S. Traikov, O. Vvedenskaya, A. Schuhmann, S. Segeletz, A. Shevchenko, A. Shevchenko

Date Published: 30th Jul 2018

Publication Type: Not specified

Powered by
 (v.1.11.1)
About FAIRDOM | LiSyM | Funding and Programmes | Credits | Terms & Conditions | Imprint
Copyright © 2008 - 2021 The University of Manchester and HITS gGmbH

LiSyM Seek uses essential cookies to keep your session open and to save if you accept this banner.

By continuing to use this site you agree to the use of cookies