Fig. 4: Label-free capabilities of SRH shows the rapid pseudo-coloring via linearly mixing protein and lipid SRS images. | Nature Communications

Fig. 4: Label-free capabilities of SRH shows the rapid pseudo-coloring via linearly mixing protein and lipid SRS images.

From: Label-free multimodal optical biopsy reveals biomolecular and morphological features of diabetic kidney tissue in 2D and 3D

Fig. 4

a SRH of large human kidney ROI. b A small ROI at subcellular resolution in which nucleoli are visible. c Confocal scanning in the Z-direction can produce 3D pseudo-colored SRH images. d Orthogonal projections of (c). e SRS and TPF images of lipids, protein, and nuclei juxtaposed to the same image slice of the 3D stack shows that multiple visualization schema are possible. f Glomerular volumes can be segmented to inform the number of nuclei within the glomerulus and in the surrounding tubule cortex. Scale bar, 200 µm. g A schematic depicting the process of obtaining confocal image planes of glomeruli with at least a hemisphere within the imaging volume. Image planes between the maximum cross-sectional area and the vanishing point are retained. h Example of a few image planes of a glomerulus with the mesangium (cyan) and total glomerulus (magenta) overlaid on the CH3 stretching images. i The image planes are used to reconstruct volumes in ImageJ and MeshMixer. j Summary of glomerular factional volume [Vv(Mes/Glom)] between control samples and DKD samples shows that mean DKD glomeruli have significantly higher mesangial volume fractions compared to control specimens (n = 5 for each group). Data presented are interquartile range box plots with median line and 1.5 IQR whiskers. Two tailed t test *p = 0.0297. ROI Region of Interest, SRH Stimulated Raman Histology.

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