## -------------------------------------------------------------------------- library(CATALYST) data(raw_data) ff <- concatFCS(raw_data) ## ----eval=FALSE------------------------------------------------------------ # normCytof(x=ff, y="dvs", k=80, plot=FALSE) ## -------------------------------------------------------------------------- data(sample_ff) sample_ff ## -------------------------------------------------------------------------- data(sample_key) head(sample_key) ## ----messages=FALSE-------------------------------------------------------- re0 <- assignPrelim(x=sample_ff, y=sample_key, verbose=FALSE) re0 ## -------------------------------------------------------------------------- # estimate separation cutoffs re <- estCutoffs(x=re0) ## ----results = 'hide'------------------------------------------------------ # use global separation cutoff applyCutoffs(x=re, sep_cutoffs=0.35) # use population-specific cutoffs re <- applyCutoffs(x=re) ## ----results='hide', message=FALSE----------------------------------------- outFCS(x=re, y=sample_ff) ## ----eval=FALSE------------------------------------------------------------ # plotYields(x=re, which=c("C1", 0), plotly=FALSE) ## ----echo=FALSE, fig.width=8, fig.height=4.5------------------------------- ps <- plotYields(x=re, which=c("C1", 0), plotly=FALSE) for (i in seq_along(ps)) assign(sprintf("p%s", i), ps[[i]]) p1; p2 ## ----fig.width=8, fig.height=4--------------------------------------------- # event plots for unassigned events # & barcode population D1 plotEvents(x=re, which=c(0, "D1"), n_events=25) ## ----warning=FALSE, fig.width=6, fig.height=6.5---------------------------- plotMahal(x=re, which="B3") ## -------------------------------------------------------------------------- # get single-stained control samples data(ss_exp) # specify mass channels stained for bc_ms <- c(139, 141:156, 158:176) # debarcode re <- assignPrelim(x=ss_exp, y=bc_ms, verbose=FALSE) re <- estCutoffs(x=re) re <- applyCutoffs(x=re) # compute spillover matrix spillMat <- computeSpillmat(x=re) ## ----fig.width=7, fig.height=7--------------------------------------------- plotSpillmat(bc_ms=bc_ms, SM=spillMat, plotly=FALSE) ## ---- message=FALSE-------------------------------------------------------- data(mp_cells) comped_flow <- compCytof(x=mp_cells, y=spillMat, method="flow") comped_nnls <- compCytof(x=mp_cells, y=spillMat, method="nnls") ## ----echo=FALSE, message=FALSE, results='hide'----------------------------- cf <- 20 cells_t <- asinh(exprs(mp_cells)/cf) comped_flow_t <- asinh(exprs(comped_flow)/cf) comped_nnls_t <- asinh(exprs(comped_nnls)/cf) ## ----echo=FALSE, fig.width=8.6, fig.height=3.2----------------------------- par(mfrow=c(1,3)) which <- c("Er167Di", "Er168Di") cols <- colorRampPalette(rev(RColorBrewer::brewer.pal(10, "Spectral"))) bw <- .2; n <- 100 smoothScatter(cells_t[, which], nrpoints=0, nbin=n, bandwidth=bw, colramp=cols, main='Uncompensated') smoothScatter(comped_flow_t[, which], nrpoints=0, nbin=n, bandwidth=bw, colramp=cols, main='Flow compensated') smoothScatter(comped_nnls_t[, which], nrpoints=0, nbin=n, bandwidth=bw, colramp=cols, main='NNLS compensated')