Supplementary Figure 10: Representative images of immunocytochemistry staining of differentiated cells that were used in robotic microscopy for survival and neurite-like process length analysis, and representative images of neurons that were subjected to neurite-like length analysis. | Nature Neuroscience

Supplementary Figure 10: Representative images of immunocytochemistry staining of differentiated cells that were used in robotic microscopy for survival and neurite-like process length analysis, and representative images of neurons that were subjected to neurite-like length analysis.

From: Developmental alterations in Huntington's disease neural cells and pharmacological rescue in cells and mice

Supplementary Figure 10

(a-h) Cells stain for ~5-15% DARPP-32 positive cells and ~1-15 % Ki67 cells. To determine percent of cells staining for DARPP-32 or Ki67, we subjected the DAPI nuclear stained images to our custom plugin in ImageJ to create an ROI for each cell. The images were processed, thresholded, and masked. These masks subjected to particle analysis that constraints size and shape to reduce the measuring of artifacts. The masked ROIs were overlayed on to the green channel images and pixels were measured for fluorescent intensity and size. The average intensity values were normalized to the size of the nuclei, and only the nuclei with a value or 0.2 or greater were counted as significant. The number of positive staining nuclei (cyan, blue channel) was divided by total number of nuclei to determine the percentage of KI-67 and DARPP-32 positive cells. MAP-2 (red, red channel) and DARPP-32 (yellow, green channel) staining (a,c, e, g) and staining for MAP-2 (red) and Ki67 (b, d, f,h), of the (a, b) control 28Q, (c, d) 46Q (c, g) 53Q and (e, f) 109Q. Scale bar is 100 μm. (i-p) Example images of cells analyzed for process length, (i) 18Qn2, (j) 28Qn6 (k) 46Qn1 (l) 46Qn10 (m) 53Qn3 (n) 53Qn5 (o) 109n4 (p) 109Qn5. Scale bar is 100 um.

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