Investigation of inter- and intra-tumoral heterogeneity of glioblastoma using TOF-SIMS

Published: 09-10-2019| Version 2 | DOI: 10.17632/ssgyybm6d3.2
Contributor:
Ksenia Anufrieva

Description

Here, we for the first time applied TOF-SIMS (Time-of-flight secondary ion mass spectrometry) to study glioma tumors and cells with 800 nm resolution. We also developed a method that allows to simultaneously detect distribution of proteins and metabolites in tumor sections using TOF-SIMS. Our results demonstrate that according to TOF-SIMS data glioma samples can be subdivided into clinically relevant groups and distinguished from the normal brain tissue.

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Tissues were embedded into paraffin and sectioned according to the standard protocol. At the day of experiment samples were subsequently washed three times with xylene, three times with 96% ethanol, once with 70% ethanol, twice with PBS and three times with water (mQ). After that samples were dried under a weak flow of argon and subjected to TOF-SIMS. In some experiments paraffin sections were stained with nanogold conjugated anti-Caveolin-1 antibodies. For that reason, at the day of experiment samples were subsequently washed three times with xylene, three times with 96% ethanol, once with 70% ethanol and twice with PBS. After microwave antigen retrieval in DakoCytomation target retrieval solution pH 6 (Dako), slides were blocked with 1% BSA in PBS and incubated with nanogold conjugated anti-Caveolin-1 antibodies overnight at 4°C. Next day, specimens were subsequently washed twice with 0.1% Triton-X100 in PBS, three times with PBS and three times with water (mQ). After that samples were dried under a weak flow of argon and subjected to TOF-SIMS.