Venkateswaran, S V; Kreuzaler, P; Maclachlan, C; McMahon, G; Greenidge, G; Collinson, L; Bunch, J; Yuneva, M (2025) A Multimodal Imaging Pipeline to Decipher Cell-Specific Metabolic Functions and Tissue Microenvironment Dynamics. Nature Protocols, 20. pp. 1678-1699.

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Abstract

In this paper we describe a multimodal imaging pipeline that allows cell type identification and nanoscale tracing of stable isotope labelled compounds. This pipeline extends upon the principles of correlative light, electron, and ion microscopy (CLEIM), by combining confocal microscopy reporter or probe-based fluorescence, electron microscopy (EM), stable isotope labelling and Nanoscale secondary ion mass spectrometry (NanoSIMS). We apply this method to a murine model of hepatocellular carcinoma to study uptake of glucose derived carbon (13C) and glutamine derived nitrogen (15N) by tumour associated immune cells. In vivo labelling with fluorescent-tagged antibodies (B220, CD3, CD8a) in tandem with confocal microscopy allows for the identification of specific cell types (B cells and T cells) in the tumour microenvironment. Subsequent image correlation with electron microscopy offers the contrast and resolution to image membranes and organelles. NanoSIMS tracks the enrichment of stable isotopes within these intracellular compartments. Our pipeline caters to a broad spectrum of applications, as it can easily be adapted to trace the uptake and utilisation of any stable isotope labelled nutrient, drug, or a probe by defined cellular populations in any tissue in situ.

Item Type:	Article
Subjects:	Biotechnology > Biopharmaceutical Manufacturing and Characterisation
Divisions:	Chemical & Biological Sciences
Identification number/DOI:	10.1038/s41596-024-01118-4
Last Modified:	06 Oct 2025 09:21
URI:	https://eprintspublications.npl.co.uk/id/eprint/10166