Analysis pipeline detects the position and characteristics of all human chromosomes in three-dimensions
(A–C) Schematic of neutrophil isolation, chromosome paint (A), spectral character of each human chromosome (B), and image analysis pipeline (C). (D) Proportion of total chromosomes detected made up by each chromosome in human blood neutrophils. Red line represents expected proportion if all 24 chromosomes were detected equally (0.04 for autosomes, 0.02 for sex chromosomes).
(E and F) Box and whisker plot (fifth-95th percentile) of the volume (μm3) (E) and surface area (μm2) (F) of each chromosome across all 240 neutrophils. Unpaired T test used to compare X and Y chromosomes.
(G) Plot of median chromosome volume across all neutrophils against chromosome linear length with straight line fitted (y = 1.09e-3x+0.649, R2 = 0.359 and P = 0.001). Dark and light gray points indicate chromosome was called as less than 0.03 or 0.02 of all chromosomes respectively, as in Figure 1D.