Fig 1: PySeq2500 automated 4i staining of adjacent spinal cord sections across flow cells and runs (A) Spinal cord sections from SOD1 G93A and nontransgenic animals stained for GFAP (astrocytes), IBA1 (microglia), and ELAVL2 (neurons) in two 4i cycles. Adjacent tissue sections from each animal were processed on flow cells run in parallel (run 1A and 1B) and across multiple experiments (run 1 and run 2). Scale bars represent 500 µm. (B) Histograms of signal intensity for each marker in manually annotated dorsal and ventral horn regions colored by experimental run.
Fig 2: Simultaneous 4 channel imaging using PySeq2500. (A) Distribution of pixel signal intensity proportion across all detection channels from each fluorophore used for 4 channel imaging. (B) Mouse spinal cord stained simultaneously for LMNB1, GFAP, MBP, and ELAVL2 with fluorophore crosstalk removed using PICASSO unmixing strategy. Scale bar represents 500 µm. (C) Inset of B. Scale bar represents 100 µm. (D) Pearson correlation coefficient, r, of pixel intensities between corresponding detection channels prior to unmixing from singleplex staining of (top) LMNB1 with an AF532 conjugated secondary antibody and (bottom) MBP with a Cy5 conjugated secondary antibody. (E) Pearson correlation coefficient of pixel intensities between corresponding detection channels after linear unmixing of signal from singlex staining of (top) LMNB1 with an AF532 conjugated secondary antibody and (bottom) MBP with a Cy5 conjugated secondary antibody.
Supplier Page from MilliporeSigma for Anti-ELAVL2 antibody produced in rabbit