An automated and fast sample preparation workflow for laser microdissection guided ultrasensitive proteomics

Item Type:	Article
Title:	An automated and fast sample preparation workflow for laser microdissection guided ultrasensitive proteomics
Creators Name:	Makhmut, A. and Qin, D. and Hartlmayr, D. and Seth, A. and Coscia, F.
Abstract:	Spatial tissue proteomics integrating whole-slide imaging, laser microdissection and ultrasensitive mass spectrometry is a powerful approach to link cellular phenotypes to functional proteome states in (patho)physiology. To be applicable to large patient cohorts and low sample input amounts, including single-cell applications, loss-minimized and streamlined end-to-end workflows are key. We here introduce an automated sample preparation protocol for laser microdissected samples utilizing the cellenONE® robotic system, which has the capacity to process 192 samples in three hours. Following laser microdissection collection directly into the proteoCHIP LF 48 or EVO 96 chip, our optimized protocol facilitates lysis, formalin de-crosslinking and tryptic digest of low-input archival tissue samples. The seamless integration with the Evosep ONE LC system by centrifugation allows 'on-the-fly' sample clean-up, particularly pertinent for laser microdissection workflows. We validate our method in human tonsil archival tissue, where we profile proteomes of spatially-defined B-cell, T-cell and epithelial microregions of 4,000 μm(2) to a depth of ∼2,000 proteins and with high cell type specificity. We finally provide detailed equipment templates and experimental guidelines for broad accessibility.
Keywords:	Animals, Mice
Source:	Molecular & Cellular Proteomics
ISSN:	1535-9484
Publisher:	American Society for Biochemistry and Molecular Biology
Page Range:	100750
Date:	20 March 2024
Official Publication:	https://doi.org/10.1016/j.mcpro.2024.100750
PubMed:	View item in PubMed

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