## ----setup, include=FALSE----------------------------------------------------- knitr::opts_chunk$set(echo = TRUE, dev = "png") ## ----load, eval=TRUE, message=FALSE------------------------------------------- library(celda) ## ----load_10X, eval=TRUE, message=FALSE--------------------------------------- # Install TENxPBMCData if is it not already if (!requireNamespace("TENxPBMCData", quietly = TRUE)) { if (!requireNamespace("BiocManager", quietly = TRUE)) { install.packages("BiocManager") } BiocManager::install("TENxPBMCData") } # Load PBMC data library(TENxPBMCData) sce <- TENxPBMCData("pbmc4k") colnames(sce) <- paste(sce$Sample, sce$Barcode, sep = "_") rownames(sce) <- rowData(sce)$Symbol_TENx ## ----decontX_background, eval=FALSE, message=FALSE---------------------------- # sce <- decontX(sce, background = raw) ## ----decontX, eval=TRUE, message=FALSE---------------------------------------- sce <- decontX(sce) ## ----UMAP_Clusters------------------------------------------------------------ umap <- reducedDim(sce, "decontX_UMAP") plotDimReduceCluster(x = sce$decontX_clusters, dim1 = umap[, 1], dim2 = umap[, 2]) ## ----plot_decon--------------------------------------------------------------- plotDecontXContamination(sce) ## ----plot_feature, message=FALSE---------------------------------------------- library(scater) sce <- logNormCounts(sce) plotDimReduceFeature(as.matrix(logcounts(sce)), dim1 = umap[, 1], dim2 = umap[, 2], features = c("CD3D", "CD3E", "GNLY", "LYZ", "S100A8", "S100A9", "CD79A", "CD79B", "MS4A1"), exactMatch = TRUE) ## ----barplotCounts------------------------------------------------------------ markers <- list(Tcell_Markers = c("CD3E", "CD3D"), Bcell_Markers = c("CD79A", "CD79B", "MS4A1"), Monocyte_Markers = c("S100A8", "S100A9", "LYZ"), NKcell_Markers = "GNLY") cellTypeMappings <- list(Tcells = 2, Bcells = 5, Monocytes = 1, NKcells = 6) plotDecontXMarkerPercentage(sce, markers = markers, groupClusters = cellTypeMappings, assayName = "counts") ## ----barplotDecontCounts------------------------------------------------------ plotDecontXMarkerPercentage(sce, markers = markers, groupClusters = cellTypeMappings, assayName = "decontXcounts") ## ----barplotBoth-------------------------------------------------------------- plotDecontXMarkerPercentage(sce, markers = markers, groupClusters = cellTypeMappings, assayName = c("counts", "decontXcounts")) ## ----plotDecontXMarkerExpression---------------------------------------------- plotDecontXMarkerExpression(sce, markers = markers[["Monocyte_Markers"]], groupClusters = cellTypeMappings, ncol = 3) ## ----plot_norm_counts, eval = FALSE------------------------------------------- # sce <- scater::logNormCounts(sce, # exprs_values = "decontXcounts", # name = "dlogcounts") # # plotDecontXMarkerExpression(sce, # markers = markers[["Monocyte_Markers"]], # groupClusters = cellTypeMappings, # ncol = 3, # assayName = c("logcounts", "dlogcounts")) ## ----findDelta---------------------------------------------------------------- metadata(sce)$decontX$estimates$all_cells$delta ## ----newDecontX, eval=TRUE, message=FALSE------------------------------------- sce.delta <- decontX(sce, delta = c(9, 20), estimateDelta = FALSE) plot(sce$decontX_contamination, sce.delta$decontX_contamination, xlab = "DecontX estimated priors", ylab = "Setting priors to estimate higher contamination") abline(0, 1, col = "red", lwd = 2) ## ----------------------------------------------------------------------------- sessionInfo()