Compiled: July 15, 2019
This vignette demonstrates the use of the Conos package in Seurat. Commands and parameters are based off of the Conos tutorial. If you use Conos in your work, please cite:
Joint analysis of heterogeneous single-cell RNA-seq dataset collections
Nikolas Barkas, Viktor Petukhov, Daria Nikolaeva, Yaroslav Lozinsky, Samuel Demharter, Konstantin Khodosevich, Peter V. Kharchenko
Nature Methods, 2019.
Prerequisites to install:
library(conos)
library(Seurat)
library(SeuratData)
library(SeuratWrappers)
To learn more about this dataset, type ?pbmcsca
InstallData("pbmcsca")
data("pbmcsca")
pbmcsca.panel <- SplitObject(pbmcsca, split.by = "Method")
for (i in 1:length(pbmcsca.panel)) {
pbmcsca.panel[[i]] <- NormalizeData(pbmcsca.panel[[i]]) %>% FindVariableFeatures() %>% ScaleData() %>%
RunPCA(verbose = FALSE)
}
pbmcsca.con <- Conos$new(pbmcsca.panel)
pbmcsca.con$buildGraph(k = 15, k.self = 5, space = "PCA", ncomps = 30, n.odgenes = 2000, matching.method = "mNN",
metric = "angular", score.component.variance = TRUE, verbose = TRUE)
pbmcsca.con$findCommunities()
pbmcsca.con$embedGraph()
pbmcsca <- as.Seurat(pbmcsca.con)
DimPlot(pbmcsca, reduction = "largeVis", group.by = c("Method", "ident", "CellType"), ncol = 3)
To learn more about this dataset, type ?ifnb
InstallData("ifnb")
data("ifnb")
ifnb.panel <- SplitObject(ifnb, split.by = "stim")
for (i in 1:length(ifnb.panel)) {
ifnb.panel[[i]] <- NormalizeData(ifnb.panel[[i]]) %>% FindVariableFeatures() %>% ScaleData() %>%
RunPCA(verbose = FALSE)
}
ifnb.con <- Conos$new(ifnb.panel)
ifnb.con$buildGraph(k = 15, k.self = 5, space = "PCA", ncomps = 30, n.odgenes = 2000, matching.method = "mNN",
metric = "angular", score.component.variance = TRUE, verbose = TRUE)
ifnb.con$findCommunities()
ifnb.con$embedGraph()
ifnb <- as.Seurat(ifnb.con)
DimPlot(ifnb, reduction = "largeVis", group.by = c("stim", "ident", "seurat_annotations"), ncol = 3)
To learn more about this dataset, type ?panc8
InstallData("panc8")
data("panc8")
panc8.panel <- SplitObject(panc8, split.by = "replicate")
for (i in 1:length(panc8.panel)) {
panc8.panel[[i]] <- NormalizeData(panc8.panel[[i]]) %>% FindVariableFeatures() %>% ScaleData() %>%
RunPCA(verbose = FALSE)
}
panc8.con <- Conos$new(panc8.panel)
panc8.con$buildGraph(k = 15, k.self = 5, space = "PCA", ncomps = 30, n.odgenes = 2000, matching.method = "mNN",
metric = "angular", score.component.variance = TRUE, verbose = TRUE)
panc8.con$findCommunities()
panc8.con$embedGraph()
panc8 <- as.Seurat(panc8.con)
DimPlot(panc8, reduction = "largeVis", group.by = c("replicate", "ident", "celltype"), ncol = 3)
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