Dual-Mode Wheat Germ Agglutinin Labeling - A Versatile Cell Segmentation Strategy for High-Resolution LA-ICP-TOFMS Bioimaging
- Author(s)
- Claude Molitor, Martin Schaier, David Loibnegger, Gabriel Braun, Michael Gutmann, Walter Berger, Gunda Koellensperger
- Abstract
Single-cell analysis by laser ablation inductively coupled plasma time-of-flight mass spectrometry (LA-ICP-TOFMS) enables high-resolution mapping of elemental distributions and cellular phenotypes. Segmentation of individual cells necessitates labeling of both nuclei and membranes, the latter often requiring extensive tissue-specific optimization. In this study, we present a broadly applicable segmentation protocol based on wheat germ agglutinin (WGA), a lectin that binds to N-acetylglucosamine and sialic acid residues ubiquitously expressed on the cell membrane. By combining fluorescently labeled WGA with a metal-tagged anti-WGA antibody, we introduce a dual-labeling strategy compatible with both fluorescence microscopy and LA-ICP-TOFMS, enabling cross-validation of membrane labeling and enhancing segmentation accuracy. With recent advancements in laser ablation technology, such as higher repetition rates and submicrometer spot sizes, high-resolution imaging across large sample areas has become increasingly feasible. The robust, high-contrast membrane labeling achieved with our method facilitates precise cell segmentation at these resolutions and enhances the quality of the downstream single-cell data analysis. Beyond that, our approach reduces staining costs, streamlines workflows, and provides a scalable alternative to existing membrane-labeling strategies.
- Organisation(s)
- Department of Inorganic Chemistry, Department of Analytical Chemistry
- External organisation(s)
- Vienna Doctoral School in Chemistry (DoSChem), Medizinische Universität Wien
- Journal
- Analytical Chemistry
- ISSN
- 0003-2700
- DOI
- https://doi.org/10.1021/acs.analchem.5c04060
- Publication date
- 09-2025
- Peer reviewed
- Yes
- Austrian Fields of Science 2012
- 104002 Analytical chemistry, 104026 Spectroscopy
- Portal url
- https://ucrisportal.univie.ac.at/en/publications/7c22bb3a-147e-4892-8f00-7773f0e6cccd