Figure 2
From: H-ABC tubulinopathy revealed by label-free second harmonic generation microscopy
The corpus callosum of taiep rats hosts cells that emit second harmonic signals. (A) Location of the imaged and analyzed region of the CC. (B–D and E–G) Representative SHG micrographs from the CC of a WT and a TUBB4A mutant rat, respectively; Z-stacks of five images, each spaced 7 μm, were generated. In (B) and (E) the sum projections of the five planes are shown, to better appreciate the continuity of the emitting structures in the corpus callosum. All animals tested gave identical results. In (C) and (F) only one plane is shown. Images (D) and (G) are high magnification images from squares in (C) and (F), respectively. Arrow, cell bodies; dotted arrow, elongated structures. (H) Gray values plotted from the line scans crossing over the fibers in the CC of WT and taiep rats; line and image sources are shown just below. (I) Gray level histogram obtained from regions of interest (ROIs) in the CC of the rats (26 + 16 images from 3 taiep and 2 WT rats). (J, K) Gray values of surface profiles; intensities obtained from elongated structures in demyelinated regions are higher than those found in similar locations in healthy rats. The calibration bar in (E) is valid for (B, C) and (F), and the calibration bar in (D) is also valid for (G), with brighter colors representing the highest signal intensity. The direction of laser polarization for SHG microscopy coincides with the scale bar orientation. The average power used for SHG imaging was 13 mW.
