Fig. 6: Defining the surrounding cellular (immune-) landscape in the model of acute myocardial infarction.

a Multi-omics approach to decipher spectra from cells by combination of Raman spectroscopy with consecutive MACSima^TM multicolor immunofluorescence staining. b H&E staining of an adjacent section of murine myocardial infarction. Scale bar 300 µm. c MACSima™ multiplexed immunofluorescence imaging was performed on the identical section and region as Raman spectroscopy was done previously. Scale bar 50 µm. n = 1 individual sample. d Raman Intensity images at 2940 cm⁻¹ (general band for lipids and proteins). Scale bar 100 µm. e Cluster image identified by the Seurat workflow. Cluster 0 was assigned to myocardium. f tSNE plot after dimensionality reduction of Raman spectra of identified cell types. Clear separation into cardiomyocytes, erythrocytes and immune cells. g Spectral subphenotyping of immune cells. Neutrophils display the largest cluster while MHC II+ professional antigen-presenting cells (pAPCs) and CD68+ macrophages separate into a distinct cluster. h Donut chart showing frequency distribution of identified cells (percent from absolute number of analyzed number of pixels). i Venn diagram of surface markers analyzed for the immune cell subpopulation. E.g., most Ly6G+ cells were also CD45 + . j Average spectra for the different cell types and characteristic peaks, together with the spatial representation of the analyzed pixels. vSMCs: vascular smooth muscle cells.