Fig. 1.

Single cell photonics from normal and injured vessels ex vivo following flow restriction. (a). Representative haematoxylin and eosin (H&E) staining of LCA from sham and ligated vessels 14 days post-ligation (A: Adventitia, M: Media, I:Intima and NI:Neointima). (b). Morphometric analysis of adventitial, medial and intimal volumes within the LCA in sham and ligated arteries; (#p≤0.05, n=3). (c). Representative immunohistochemical analysis of Myh11 (red) and α-actin expression (red) and DAPI nuclei staining (blue) in adventitial, medial and intimal layers of the LCA from sham and ligated carotid arteries. (d). Representative immunoblot of Myh11 expression in the LCA from sham and ligated carotid artery (n=3). (e). Schematic of capture of individual cells using the LoaD platform. (f). Representative image of formalin-fixed cells from sham and ligated vessels captured on a V-cup using the LoaD platform. (g). Single cell auto-fluorescence photon emissions across five broadband light wavelengths (λ465, λ530, λ565, λ630 and λ670 nm with a bandwidth of 20 nm) from sham and ligated LCA ex vivo. Data are the mean ± SEM of Log10 photons from 178 cells/group from 6 pooled vessels, #p≤0.001 vs sham. (h). Log2 fold increase over background of single cell photon emissions from sham and ligated LCA. Data are the mean ± SEM of 178 cells/group from 6 pooled vessels, #p≤0.001 vs sham. (i). PCA biplots of single cell photon analysis from sham (black) and ligated (orange) vessels. Each λ variable is representative of the eigenvectors (loadings). (j). PCA loading plots of single cell photon analysis from sham (black) and ligated (orange) cells ex vivo compared to Ramos B cells in vitro (red). Data are from 411 cells across the five wavelengths. (k). LDA plots of single cell photons from sham (black) and ligated (orange) vessels. (l). Confusion matrix of true class representing the given group assignment and predicted class representing the estimated group assignment following a leave-one-outcross-validation procedure