Assessment of different sample preparation routes for mass spectrometric monitoring and imaging of lipids in bone cells via ToF-SIMS

Biointerphases
Kaija SchaepeJürgen Janek

Abstract

In ToF-SIMS analysis, the experimental outcome from cell experiments is to a great extent influenced by the sample preparation routine. In order to better judge this critical influence in the case of lipid analysis, a detailed comparison of different sample preparation routines is performed-aiming at an optimized preparation routine for systematic lipid imaging of cell cultures. For this purpose, human mesenchymal stem cells were analyzed: (a) as chemically fixed, (b) freeze-dried, and (c) frozen-hydrated. For chemical fixation, different fixatives, i.e., glutaraldehyde, paraformaldehyde, and a mixture of both, were tested with different postfixative handling procedures like storage in phosphate buffered saline, water or critical point drying. Furthermore, secondary lipid fixation via osmium tetroxide was taken into account and the effect of an ascending alcohol series with and without this secondary lipid fixation was evaluated. Concerning freeze-drying, three different postprocessing possibilities were examined. One can be considered as a pure cryofixation technique while the other two routes were based on chemical fixation. Cryofixation methods known from literature, i.e., freeze-fracturing and simple frozen-hydrated prepara...Continue Reading

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Citations

Jan 27, 2016·Biointerphases·Kaija SchaepeJürgen Janek
Jun 19, 2016·Analytical Biochemistry·Justyna BobrowskaMalgorzata Lekka
Nov 25, 2016·Nature Reviews. Rheumatology·Beatriz RochaFrancisco J Blanco
May 19, 2017·Analytical and Bioanalytical Chemistry·Kaija SchaepeJürgen Janek
Apr 13, 2019·Scientific Reports·Thomas M TiefenboeckKambiz Sarahrudi
May 23, 2020·Biointerphases·Christine KernMarcus Rohnke
Jun 4, 2018·Cancer Metastasis Reviews·Alfredo Erazo-OliverasRobert S Chapkin
Nov 19, 2019·Analytical and Bioanalytical Chemistry·Alexander GulinAlexander Pogorelov
Aug 11, 2021·Journal of the American Society for Mass Spectrometry·Michael J TaylorChristopher R Anderton
Jul 3, 2020·Analytical Chemistry·Wenchao WeiXiao-Ying Yu

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