False-color Raman images of CD4+T cells and CD8+ T cells from control and LPS-treated animals after day 1, 4, 10, and 30 obtained by k-means cluster analysis.

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False-color Raman images of CD4+T cells and CD8+ T cells from control and LPS-treated animals after day 1, 4, 10, and 30 obtained by k-means cluster analysis. False-color Raman images of CD4+T cells and CD8+ T cells from control and LPS-treated animals after day 1, 4, 10, and 30 obtained by k-means cluster analysis. (A) Representative false-color Raman images of CD4+ T cells and CD8+ T cells from control and LPS-treated animals after day 1, 4, 10, and 30 obtained by applying k-means clustering method. The x-axis and y-axis show step size (0.3 μm) used for Raman imaging. The k-means algorithm was executed to find four different clusters of spectra for each time point in the cells recovered from spleen of control and LPS-treated animals. The four clusters are represented by blue, cyan, green, and red color in the image. Blue cluster represents the narrow cytoplasm region of the T cell, cyan and green cluster highlight the nucleus, and red cluster is a mixture of the outer parts of the cell and remaining background contributions. The contribution of each cluster has been represented on right side of individual cells as cluster scores. (B) Corresponding Raman spectra of the four clusters obtained from k-means analysis using the same color code as in (A). Anuradha Ramoji et al. ImmunoHorizons 2019;3:45-60 Copyright © 2019 The Authors