## ---- include = FALSE--------------------------------------------------------- knitr::opts_chunk$set( collapse = TRUE, comment = "#>" ) ## ----setup, eval=FALSE-------------------------------------------------------- # library(igraph) # library(schex) # library(TENxPBMCData) # library(scater) # library(scran) # library(ggrepel) ## ----load, eval=FALSE--------------------------------------------------------- # tenx_pbmc3k <- TENxPBMCData(dataset = "pbmc3k") # # rownames(tenx_pbmc3k) <- uniquifyFeatureNames(rowData(tenx_pbmc3k)$ENSEMBL_ID, # rowData(tenx_pbmc3k)$Symbol_TENx) ## ----filter-cells, eval=FALSE------------------------------------------------- # rowData(tenx_pbmc3k)$Mito <- grepl("^MT-", rownames(tenx_pbmc3k)) # colData(tenx_pbmc3k) <- cbind(colData(tenx_pbmc3k), # perCellQCMetrics(tenx_pbmc3k, # subsets=list(Mt=rowData(tenx_pbmc3k)$Mito))) # rowData(tenx_pbmc3k) <- cbind(rowData(tenx_pbmc3k), # perFeatureQCMetrics(tenx_pbmc3k)) # # tenx_pbmc3k <- tenx_pbmc3k[, !colData(tenx_pbmc3k)$subsets_Mt_percent > 50] # # libsize_drop <- isOutlier(tenx_pbmc3k$total, # nmads = 3,type = "lower", log = TRUE) # feature_drop <- isOutlier(tenx_pbmc3k$detected, # nmads = 3, type = "lower", log = TRUE) # # tenx_pbmc3k <- tenx_pbmc3k[, !(libsize_drop | feature_drop)] ## ----filter-genes, eval=FALSE------------------------------------------------- # rm_ind <- calculateAverage(tenx_pbmc3k)<0 # tenx_pbmc3k <- tenx_pbmc3k[!rm_ind,] ## ----norm, message=FALSE, warning=FALSE, eval=FALSE--------------------------- # tenx_pbmc3k <- scater::logNormCounts(tenx_pbmc3k) ## ----dim-red, message=FALSE, warning=FALSE, eval=FALSE------------------------ # tenx_pbmc3k <- runPCA(tenx_pbmc3k) # set.seed(10) # tenx_pbmc3k <- runUMAP(tenx_pbmc3k, dimred = "PCA", spread = 1, # min_dist = 0.4) ## ----cluster, eval=FALSE------------------------------------------------------ # snn_gr <- buildSNNGraph(tenx_pbmc3k, use.dimred = "PCA", k = 50) # clusters <- cluster_louvain(snn_gr) # tenx_pbmc3k$cluster <- factor(clusters$membership) ## ---- eval=FALSE-------------------------------------------------------------- # plot_hexbin_density_shiny(tenx_pbmc3k, 10, 50, dimension_reduction = "UMAP") ## ---- eval=FALSE-------------------------------------------------------------- # plot_hexbin_feature_shiny(tenx_pbmc3k, type="counts", feature="POMGNT1", # action="prop_0", min_nbins=10, max_nbins=50, dimension_reduction="UMAP", # mod="RNA") ## ---- eval=FALSE-------------------------------------------------------------- # plot_hexbin_meta_shiny(tenx_pbmc3k, col="cluster", # action="majority", min_nbins=10, max_nbins=50, dimension_reduction="UMAP")