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Article Dans Une Revue Analytical Chemistry Année : 2021

Toward high spatially resolved proteomics using expansion microscopy

Résumé

Expansion microscopy (EM) is an emerging approach for morphological examination of biological specimens at nanoscale resolution using conventional optical microscopy. To achieve physical separation of cell structures, tissues are embedded in a swellable polymer and expanded several fold in an isotropic manner. This work shows the development and optimization of physical tissue expansion as a new method for spatially resolved large-scale proteomics. Herein we established a novel method to enlarge the tissue section to be compatible with manual microdissection on regions of interest and MS-based proteomic analysis. A major issue in expansion microscopy is the loss of protein information during the mechanical homogenization phase due to the use of proteinase K. For isotropic expansion, different homogenization agents were investigated, both to maximize protein identification and to minimize protein diffusion. Best results were obtained with SDS for homogenization. Using our modified protocol, we were able to enlarge a tissue section more than 3-fold and identified up to 655 proteins from 1 mm in size after expansion, equivalent to 330 mum in their real size corresponding thus to an average of 260 cells. This approach can be performed easily without any expensive sampling instrument. We demonstrated the compatibility of sample preparation for expansion microscopy and proteomic study in a spatial context.
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Dates et versions

hal-04011475 , version 1 (02-03-2023)

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Lauranne Drelich, Soulaimane Aboulouard, Julien Franck, Michel Salzet, Isabelle Fournier, et al.. Toward high spatially resolved proteomics using expansion microscopy. Analytical Chemistry, 2021, Analytical Chemistry, 93 (36), pp.12195-12203. ⟨10.1021/acs.analchem.0c05372⟩. ⟨hal-04011475⟩

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