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Add overlay plot with ggplot2

Mike Jiang
04/13/2015

ggcyto makes ggplot2 to be able to work with Cytometry data, namely flowSet/ncdfFlowSet or flowFrame S4 objects.

library(ggcyto)
data(GvHD)
fs <- GvHD[subset(pData(GvHD), Patient %in%5:7 & Visit %in% c(5:6))[["name"]]]
fr <- fs[[1]]

add other gated data as the overlays

p <- ggplot(fr, aes(x = `FSC-H`, y =  `SSC-H`))

# change the smooth color 
myColor <- rev(RColorBrewer::brewer.pal(11, "Spectral"))
myColor_scale_fill <- scale_fill_gradientn(colours = myColor)
# get a gated data
lg <- flowStats::lymphGate(fr, channels=c("FSC-H", "SSC-H"),scale=0.6)
norm.filter <- lg$n2gate
overlay.fr <- lg$x

# add the overlay and dim the background
p1 <- p + stat_binhex(bin = 64, alpha = 0.2)
p1 <- p1 + geom_point(data = overlay.fr, color = "red", cex = 0.5, pch = 19)
p1

# another overlay 
rect.g <- rectangleGate(list("FSC-H" =  c(300,500), "SSC-H" = c(50,200)))
overlay.fr2 <- Subset(fr, rect.g)
p1 + geom_point(data = overlay.fr2, color = "black", cex = 0.5, pch = 19)

# overlay for multiple samples
overlay.fs1 <- Subset(fs, norm.filter)
overlay.fs2 <- Subset(fs, rect.g)
ggplot(fs, aes(x = `FSC-H`, y =  `SSC-H`)) + 
          scale_fill_gradientn(colours = myColor)  +
          stat_binhex(bin = 64, alpha = 0.2) +
          geom_point(data = overlay.fs1, color = "red", cex = 0.5, pch = 19) +
          geom_point(data = overlay.fs2, color = "black", cex = 0.5, pch = 19) +
          facet_grid(Patient ~ Visit)