Figure 1
From: Multicolored stain-free histopathology with coherent Raman imaging
Stain-free histologic imaging with multicolor coherent Raman imaging. (a) Vibrational spectra of the major constituents of tissue: lipids, protein and water. Arrows indicate Raman shifts at which imaging is performed. (b–d) stimulated Raman scattering (SRS) images of a live C2C12 mammalian cell acquired at the CH2-stretching vibration at 2845 cm−1 (b) and CH3-stretching vibration at 2940 cm−1 (c). Multicolor image (d) generated from images (b, c) with the green channel (CH2 image) showing the cell-body and the blue channel (thresholded CH3-CH2 difference image) highlighting the nuclear morphology including a bright nucleolus. (e–h) SRS images of fresh ex vivo brain tissue acquired at CH2-stretching vibration at 2845 cm−1 (e), CH3-stretching vibration at 2940 cm−1 (f), and vibrationally off-resonant showing two-color two-photon absorption of hemoglobin at a sum frequency of 23 700 cm−1 (g). Multicolor image (h) generated from images (e–g) with the green channel (CH2 image) highlighting cytoplasm and myelin sheaths, blue channel (thresholded CH3-CH2 difference image) showing the nuclear morphology, and the red channel (hemoglobin image) highlighting the red blood cells. (i) Hematoxylin & eosin (H&E)-stained micrograph from the same region in the brain. (j) Same multicolor image as (h) with a different pseudo-color scheme, chosen to mimic the appearance of an H&E-stained micrograph, illustrates the similar image content and appearance of stain-free images and H&E-stained sections. Scale bar, 25 μm.
