Introduction
tidySingleCellExperiment provides a bridge between Bioconductor single-cell packages [@amezquita2019orchestrating] and the tidyverse [@wickham2019welcome]. It creates an invisible layer that enables viewing the Bioconductor SingleCellExperiment object as a tidyverse tibble, and provides SingleCellExperiment-compatible dplyr, tidyr, ggplot and plotly functions. This allows users to get the best of both Bioconductor and tidyverse worlds.
Functions/utilities available
| SingleCellExperiment-compatible Functions | Description |
|---|---|
all |
After all tidySingleCellExperiment is a SingleCellExperiment object, just better |
| tidyverse Packages | Description |
|---|---|
dplyr |
All dplyr tibble functions (e.g. tidySingleCellExperiment::select) |
tidyr |
All tidyr tibble functions (e.g. tidySingleCellExperiment::pivot_longer) |
ggplot2 |
ggplot (tidySingleCellExperiment::ggplot) |
plotly |
plot_ly (tidySingleCellExperiment::plot_ly) |
| Utilities | Description |
|---|---|
tidy |
Add tidySingleCellExperiment invisible layer over a SingleCellExperiment object |
as_tibble |
Convert cell-wise information to a tbl_df |
join_features |
Add feature-wise information, returns a tbl_df |
Installation
if (!requireNamespace("BiocManager", quietly=TRUE))
install.packages("BiocManager")
BiocManager::install("tidySingleCellExperiment")
Load libraries used in this vignette.
# Bioconductor single-cell packages
library(scater)
library(scran)
library(SingleR)
library(SingleCellSignalR)
# Tidyverse-compatible packages
library(ggplot2)
library(purrr)
library(tidyHeatmap)
# Both
library(tidySingleCellExperiment)
Create tidySingleCellExperiment, the best of both worlds!
This is a SingleCellExperiment object but it is evaluated as a tibble. So it is compatible both with SingleCellExperiment and tidyverse.
pbmc_small_tidy <- tidySingleCellExperiment::pbmc_small
It looks like a tibble
pbmc_small_tidy
## # A SingleCellExperiment-tibble abstraction: 80 × 17
## # [90mFeatures=230 | Cells=80 | Assays=counts, logcounts[0m
## .cell orig.ident nCount_RNA nFeature_RNA RNA_snn_res.0.8 letter.idents groups
## <chr> <fct> <dbl> <int> <fct> <fct> <chr>
## 1 ATGC… SeuratPro… 70 47 0 A g2
## 2 CATG… SeuratPro… 85 52 0 A g1
## 3 GAAC… SeuratPro… 87 50 1 B g2
## 4 TGAC… SeuratPro… 127 56 0 A g2
## 5 AGTC… SeuratPro… 173 53 0 A g2
## 6 TCTG… SeuratPro… 70 48 0 A g1
## 7 TGGT… SeuratPro… 64 36 0 A g1
## 8 GCAG… SeuratPro… 72 45 0 A g1
## 9 GATA… SeuratPro… 52 36 0 A g1
## 10 AATG… SeuratPro… 100 41 0 A g1
## # ℹ 70 more rows
## # ℹ 10 more variables: RNA_snn_res.1 <fct>, file <chr>, ident <fct>,
## # PC_1 <dbl>, PC_2 <dbl>, PC_3 <dbl>, PC_4 <dbl>, PC_5 <dbl>, tSNE_1 <dbl>,
## # tSNE_2 <dbl>
But it is a SingleCellExperiment object after all
assay(pbmc_small_tidy, "counts")[1:5, 1:5]
## 5 x 5 sparse Matrix of class "dgCMatrix"
## ATGCCAGAACGACT CATGGCCTGTGCAT GAACCTGATGAACC TGACTGGATTCTCA
## MS4A1 . . . .
## CD79B 1 . . .
## CD79A . . . .
## HLA-DRA . 1 . .
## TCL1A . . . .
## AGTCAGACTGCACA
## MS4A1 .
## CD79B .
## CD79A .
## HLA-DRA 1
## TCL1A .
Annotation polishing
We may have a column that contains the directory each run was taken from, such as the “file” column in pbmc_small_tidy.
pbmc_small_tidy$file[1:5]
## [1] "../data/sample2/outs/filtered_feature_bc_matrix/"
## [2] "../data/sample1/outs/filtered_feature_bc_matrix/"
## [3] "../data/sample2/outs/filtered_feature_bc_matrix/"
## [4] "../data/sample2/outs/filtered_feature_bc_matrix/"
## [5] "../data/sample2/outs/filtered_feature_bc_matrix/"
We may want to extract the run/sample name out of it into a separate column. Tidyverse extract can be used to convert a character column into multiple columns using regular expression groups.
# Create sample column
pbmc_small_polished <-
pbmc_small_tidy %>%
extract(file, "sample", "../data/([a-z0-9]+)/outs.+", remove=FALSE)
# Reorder to have sample column up front
pbmc_small_polished %>%
select(sample, everything())
## # A SingleCellExperiment-tibble abstraction: 80 × 18
## # [90mFeatures=230 | Cells=80 | Assays=counts, logcounts[0m
## .cell sample orig.ident nCount_RNA nFeature_RNA RNA_snn_res.0.8 letter.idents
## <chr> <chr> <fct> <dbl> <int> <fct> <fct>
## 1 ATGC… sampl… SeuratPro… 70 47 0 A
## 2 CATG… sampl… SeuratPro… 85 52 0 A
## 3 GAAC… sampl… SeuratPro… 87 50 1 B
## 4 TGAC… sampl… SeuratPro… 127 56 0 A
## 5 AGTC… sampl… SeuratPro… 173 53 0 A
## 6 TCTG… sampl… SeuratPro… 70 48 0 A
## 7 TGGT… sampl… SeuratPro… 64 36 0 A
## 8 GCAG… sampl… SeuratPro… 72 45 0 A
## 9 GATA… sampl… SeuratPro… 52 36 0 A
## 10 AATG… sampl… SeuratPro… 100 41 0 A
## # ℹ 70 more rows
## # ℹ 11 more variables: groups <chr>, RNA_snn_res.1 <fct>, file <chr>,
## # ident <fct>, PC_1 <dbl>, PC_2 <dbl>, PC_3 <dbl>, PC_4 <dbl>, PC_5 <dbl>,
## # tSNE_1 <dbl>, tSNE_2 <dbl>
Preliminary plots
Set colours and theme for plots.
# Use colourblind-friendly colours
friendly_cols <- dittoSeq::dittoColors()
# Set theme
custom_theme <-
list(
scale_fill_manual(values=friendly_cols),
scale_color_manual(values=friendly_cols),
theme_bw() +
theme(
panel.border=element_blank(),
axis.line=element_line(),
panel.grid.major=element_line(size=0.2),
panel.grid.minor=element_line(size=0.1),
text=element_text(size=12),
legend.position="bottom",
aspect.ratio=1,
strip.background=element_blank(),
axis.title.x=element_text(margin=margin(t=10, r=10, b=10, l=10)),
axis.title.y=element_text(margin=margin(t=10, r=10, b=10, l=10))
)
)
We can treat pbmc_small_polished as a tibble for plotting.
Here we plot number of features per cell.
pbmc_small_polished %>%
tidySingleCellExperiment::ggplot(aes(nFeature_RNA, fill=groups)) +
geom_histogram() +
custom_theme
Here we plot total features per cell.
pbmc_small_polished %>%
tidySingleCellExperiment::ggplot(aes(groups, nCount_RNA, fill=groups)) +
geom_boxplot(outlier.shape=NA) +
geom_jitter(width=0.1) +
custom_theme
Here we plot abundance of two features for each group.
pbmc_small_polished %>%
join_features(features=c("HLA-DRA", "LYZ")) %>%
ggplot(aes(groups, .abundance_counts + 1, fill=groups)) +
geom_boxplot(outlier.shape=NA) +
geom_jitter(aes(size=nCount_RNA), alpha=0.5, width=0.2) +
scale_y_log10() +
custom_theme
Preprocess the dataset
We can also treat pbmc_small_polished as a SingleCellExperiment object and proceed with data processing with Bioconductor packages, such as scran [@lun2016pooling] and scater [@mccarthy2017scater].
# Identify variable genes with scran
variable_genes <-
pbmc_small_polished %>%
modelGeneVar() %>%
getTopHVGs(prop=0.1)
# Perform PCA with scater
pbmc_small_pca <-
pbmc_small_polished %>%
runPCA(subset_row=variable_genes)
pbmc_small_pca
## # A SingleCellExperiment-tibble abstraction: 80 × 18
## # [90mFeatures=230 | Cells=80 | Assays=counts, logcounts[0m
## .cell orig.ident nCount_RNA nFeature_RNA RNA_snn_res.0.8 letter.idents groups
## <chr> <fct> <dbl> <int> <fct> <fct> <chr>
## 1 ATGC… SeuratPro… 70 47 0 A g2
## 2 CATG… SeuratPro… 85 52 0 A g1
## 3 GAAC… SeuratPro… 87 50 1 B g2
## 4 TGAC… SeuratPro… 127 56 0 A g2
## 5 AGTC… SeuratPro… 173 53 0 A g2
## 6 TCTG… SeuratPro… 70 48 0 A g1
## 7 TGGT… SeuratPro… 64 36 0 A g1
## 8 GCAG… SeuratPro… 72 45 0 A g1
## 9 GATA… SeuratPro… 52 36 0 A g1
## 10 AATG… SeuratPro… 100 41 0 A g1
## # ℹ 70 more rows
## # ℹ 11 more variables: RNA_snn_res.1 <fct>, file <chr>, sample <chr>,
## # ident <fct>, PC1 <dbl>, PC2 <dbl>, PC3 <dbl>, PC4 <dbl>, PC5 <dbl>,
## # tSNE_1 <dbl>, tSNE_2 <dbl>
If a tidyverse-compatible package is not included in the tidySingleCellExperiment collection, we can use as_tibble to permanently convert tidySingleCellExperiment into a tibble.
# Create pairs plot with GGally
pbmc_small_pca %>%
as_tibble() %>%
select(contains("PC"), everything()) %>%
GGally::ggpairs(columns=1:5, ggplot2::aes(colour=groups)) +
custom_theme
Identify clusters
We can proceed with cluster identification with scran.
pbmc_small_cluster <- pbmc_small_pca
# Assign clusters to the 'colLabels' of the SingleCellExperiment object
colLabels(pbmc_small_cluster) <-
pbmc_small_pca %>%
buildSNNGraph(use.dimred="PCA") %>%
igraph::cluster_walktrap() %$%
membership %>%
as.factor()
# Reorder columns
pbmc_small_cluster %>% select(label, everything())
## # A SingleCellExperiment-tibble abstraction: 80 × 19
## # [90mFeatures=230 | Cells=80 | Assays=counts, logcounts[0m
## .cell label orig.ident nCount_RNA nFeature_RNA RNA_snn_res.0.8 letter.idents
## <chr> <fct> <fct> <dbl> <int> <fct> <fct>
## 1 ATGCC… 2 SeuratPro… 70 47 0 A
## 2 CATGG… 2 SeuratPro… 85 52 0 A
## 3 GAACC… 2 SeuratPro… 87 50 1 B
## 4 TGACT… 1 SeuratPro… 127 56 0 A
## 5 AGTCA… 2 SeuratPro… 173 53 0 A
## 6 TCTGA… 2 SeuratPro… 70 48 0 A
## 7 TGGTA… 1 SeuratPro… 64 36 0 A
## 8 GCAGC… 2 SeuratPro… 72 45 0 A
## 9 GATAT… 2 SeuratPro… 52 36 0 A
## 10 AATGT… 2 SeuratPro… 100 41 0 A
## # ℹ 70 more rows
## # ℹ 12 more variables: groups <chr>, RNA_snn_res.1 <fct>, file <chr>,
## # sample <chr>, ident <fct>, PC1 <dbl>, PC2 <dbl>, PC3 <dbl>, PC4 <dbl>,
## # PC5 <dbl>, tSNE_1 <dbl>, tSNE_2 <dbl>
And interrogate the output as if it was a regular tibble.
# Count number of cells for each cluster per group
pbmc_small_cluster %>%
tidySingleCellExperiment::count(groups, label)
## # A tibble: 8 × 3
## groups label n
## <chr> <fct> <int>
## 1 g1 1 12
## 2 g1 2 14
## 3 g1 3 14
## 4 g1 4 4
## 5 g2 1 10
## 6 g2 2 11
## 7 g2 3 10
## 8 g2 4 5
We can identify and visualise cluster markers combining SingleCellExperiment, tidyverse functions and tidyHeatmap [@mangiola2020tidyheatmap]
# Identify top 10 markers per cluster
marker_genes <-
pbmc_small_cluster %>%
findMarkers(groups=pbmc_small_cluster$label) %>%
as.list() %>%
map(~ .x %>%
head(10) %>%
rownames()) %>%
unlist()
# Plot heatmap
pbmc_small_cluster %>%
join_features(features=marker_genes) %>%
group_by(label) %>%
heatmap(.feature, .cell, .abundance_counts, .scale="column")
Reduce dimensions
We can calculate the first 3 UMAP dimensions using the SingleCellExperiment framework and scater.
pbmc_small_UMAP <-
pbmc_small_cluster %>%
runUMAP(ncomponents=3)
And we can plot the result in 3D using plotly.
pbmc_small_UMAP %>%
plot_ly(
x=~`UMAP1`,
y=~`UMAP2`,
z=~`UMAP3`,
color=~label,
colors=friendly_cols[1:4]
)
Cell type prediction
We can infer cell type identities using SingleR [@aran2019reference] and manipulate the output using tidyverse.
# Get cell type reference data
blueprint <- celldex::BlueprintEncodeData()
# Infer cell identities
cell_type_df <-
assays(pbmc_small_UMAP)$logcounts %>%
Matrix::Matrix(sparse = TRUE) %>%
SingleR::SingleR(
ref = blueprint,
labels = blueprint$label.main,
method = "single"
) %>%
as.data.frame() %>%
as_tibble(rownames="cell") %>%
select(cell, first.labels)
# Join UMAP and cell type info
pbmc_small_cell_type <-
pbmc_small_UMAP %>%
left_join(cell_type_df, by="cell")
# Reorder columns
pbmc_small_cell_type %>%
tidySingleCellExperiment::select(cell, first.labels, everything())
## # A SingleCellExperiment-tibble abstraction: 80 × 23
## # [90mFeatures=230 | Cells=80 | Assays=counts, logcounts[0m
## cell first.labels orig.ident nCount_RNA nFeature_RNA RNA_snn_res.0.8
## <chr> <chr> <fct> <dbl> <int> <fct>
## 1 ATGCCAGAACGA… CD4+ T-cells SeuratPro… 70 47 0
## 2 CATGGCCTGTGC… CD8+ T-cells SeuratPro… 85 52 0
## 3 GAACCTGATGAA… CD8+ T-cells SeuratPro… 87 50 1
## 4 TGACTGGATTCT… CD4+ T-cells SeuratPro… 127 56 0
## 5 AGTCAGACTGCA… CD4+ T-cells SeuratPro… 173 53 0
## 6 TCTGATACACGT… CD4+ T-cells SeuratPro… 70 48 0
## 7 TGGTATCTAAAC… CD4+ T-cells SeuratPro… 64 36 0
## 8 GCAGCTCTGTTT… CD4+ T-cells SeuratPro… 72 45 0
## 9 GATATAACACGC… CD4+ T-cells SeuratPro… 52 36 0
## 10 AATGTTGACAGT… CD4+ T-cells SeuratPro… 100 41 0
## # ℹ 70 more rows
## # ℹ 17 more variables: letter.idents <fct>, groups <chr>, RNA_snn_res.1 <fct>,
## # file <chr>, sample <chr>, ident <fct>, label <fct>, PC1 <dbl>, PC2 <dbl>,
## # PC3 <dbl>, PC4 <dbl>, PC5 <dbl>, tSNE_1 <dbl>, tSNE_2 <dbl>, UMAP1 <dbl>,
## # UMAP2 <dbl>, UMAP3 <dbl>
We can easily summarise the results. For example, we can see how cell type classification overlaps with cluster classification.
# Count number of cells for each cell type per cluster
pbmc_small_cell_type %>%
count(label, first.labels)
## # A tibble: 11 × 3
## label first.labels n
## <fct> <chr> <int>
## 1 1 CD4+ T-cells 2
## 2 1 CD8+ T-cells 8
## 3 1 NK cells 12
## 4 2 B-cells 10
## 5 2 CD4+ T-cells 6
## 6 2 CD8+ T-cells 2
## 7 2 Macrophages 1
## 8 2 Monocytes 6
## 9 3 Macrophages 1
## 10 3 Monocytes 23
## 11 4 Erythrocytes 9
We can easily reshape the data for building information-rich faceted plots.
pbmc_small_cell_type %>%
# Reshape and add classifier column
pivot_longer(
cols=c(label, first.labels),
names_to="classifier", values_to="label"
) %>%
# UMAP plots for cell type and cluster
ggplot(aes(UMAP1, UMAP2, color=label)) +
geom_point() +
facet_wrap(~classifier) +
custom_theme
We can easily plot gene correlation per cell category, adding multi-layer annotations.
pbmc_small_cell_type %>%
# Add some mitochondrial abundance values
mutate(mitochondrial=rnorm(dplyr::n())) %>%
# Plot correlation
join_features(features=c("CST3", "LYZ"), shape="wide") %>%
ggplot(aes(CST3 + 1, LYZ + 1, color=groups, size=mitochondrial)) +
geom_point() +
facet_wrap(~first.labels, scales="free") +
scale_x_log10() +
scale_y_log10() +
custom_theme
Nested analyses
A powerful tool we can use with tidySingleCellExperiment is tidyverse nest. We can easily perform independent analyses on subsets of the dataset. First we classify cell types into lymphoid and myeloid, and then nest based on the new classification.
pbmc_small_nested <-
pbmc_small_cell_type %>%
filter(first.labels != "Erythrocytes") %>%
mutate(cell_class=dplyr::if_else(`first.labels` %in% c("Macrophages", "Monocytes"), "myeloid", "lymphoid")) %>%
nest(data=-cell_class)
pbmc_small_nested
## # A tibble: 2 × 2
## cell_class data
## <chr> <list>
## 1 lymphoid <SnglCllE[,40]>
## 2 myeloid <SnglCllE[,31]>
Now we can independently for the lymphoid and myeloid subsets (i) find variable features, (ii) reduce dimensions, and (iii) cluster using both tidyverse and SingleCellExperiment seamlessly.
pbmc_small_nested_reanalysed <-
pbmc_small_nested %>%
mutate(data=map(
data, ~ {
.x <- runPCA(.x, subset_row=variable_genes)
variable_genes <-
.x %>%
modelGeneVar() %>%
getTopHVGs(prop=0.3)
colLabels(.x) <-
.x %>%
buildSNNGraph(use.dimred="PCA") %>%
igraph::cluster_walktrap() %$%
membership %>%
as.factor()
.x %>% runUMAP(ncomponents=3)
}
))
pbmc_small_nested_reanalysed
## # A tibble: 2 × 2
## cell_class data
## <chr> <list>
## 1 lymphoid <SnglCllE[,40]>
## 2 myeloid <SnglCllE[,31]>
We can then unnest and plot the new classification.
pbmc_small_nested_reanalysed %>%
# Convert to tibble otherwise SingleCellExperiment drops reduced dimensions when unifying data sets.
mutate(data=map(data, ~ .x %>% as_tibble())) %>%
unnest(data) %>%
# Define unique clusters
unite("cluster", c(cell_class, label), remove=FALSE) %>%
# Plotting
ggplot(aes(UMAP1, UMAP2, color=cluster)) +
geom_point() +
facet_wrap(~cell_class) +
custom_theme
We can perform a large number of functional analyses on data subsets. For example, we can identify intra-sample cell-cell interactions using SingleCellSignalR [@cabello2020singlecellsignalr], and then compare whether interactions are stronger or weaker across conditions. The code below demonstrates how this analysis could be performed. It won’t work with this small example dataset as we have just two samples (one for each condition). But some example output is shown below and you can imagine how you can use tidyverse on the output to perform t-tests and visualisation.
pbmc_small_nested_interactions <-
pbmc_small_nested_reanalysed %>%
# Unnest based on cell category
unnest(data) %>%
# Create unambiguous clusters
mutate(integrated_clusters=first.labels %>% as.factor() %>% as.integer()) %>%
# Nest based on sample
tidySingleCellExperiment::nest(data=-sample) %>%
tidySingleCellExperiment::mutate(interactions=map(data, ~ {
# Produce variables. Yuck!
cluster <- colData(.x)$integrated_clusters
data <- data.frame(assays(.x) %>% as.list() %>% .[[1]] %>% as.matrix())
# Ligand/Receptor analysis using SingleCellSignalR
data %>%
cell_signaling(genes=rownames(data), cluster=cluster) %>%
inter_network(data=data, signal=., genes=rownames(data), cluster=cluster) %$%
`individual-networks` %>%
map_dfr(~ bind_rows(as_tibble(.x)))
}))
pbmc_small_nested_interactions %>%
select(-data) %>%
unnest(interactions)
If the dataset was not so small, and interactions could be identified, you would see something like below.
tidySingleCellExperiment::pbmc_small_nested_interactions
## # A tibble: 100 × 9
## sample ligand receptor ligand.name receptor.name origin destination
## <chr> <chr> <chr> <chr> <chr> <chr> <chr>
## 1 sample1 cluster 1.PTMA cluster… PTMA VIPR1 clust… cluster 2
## 2 sample1 cluster 1.B2M cluster… B2M KLRD1 clust… cluster 2
## 3 sample1 cluster 1.IL16 cluster… IL16 CD4 clust… cluster 2
## 4 sample1 cluster 1.HLA-B cluster… HLA-B KLRD1 clust… cluster 2
## 5 sample1 cluster 1.CALM1 cluster… CALM1 VIPR1 clust… cluster 2
## 6 sample1 cluster 1.HLA-E cluster… HLA-E KLRD1 clust… cluster 2
## 7 sample1 cluster 1.GNAS cluster… GNAS VIPR1 clust… cluster 2
## 8 sample1 cluster 1.B2M cluster… B2M HFE clust… cluster 2
## 9 sample1 cluster 1.PTMA cluster… PTMA VIPR1 clust… cluster 3
## 10 sample1 cluster 1.CALM1 cluster… CALM1 VIPR1 clust… cluster 3
## # ℹ 90 more rows
## # ℹ 2 more variables: interaction.type <chr>, LRscore <dbl>
Session Info
sessionInfo()
## R version 4.3.0 RC (2023-04-13 r84269)
## Platform: x86_64-pc-linux-gnu (64-bit)
## Running under: Ubuntu 22.04.2 LTS
##
## Matrix products: default
## BLAS: /home/biocbuild/bbs-3.17-bioc/R/lib/libRblas.so
## LAPACK: /usr/lib/x86_64-linux-gnu/lapack/liblapack.so.3.10.0
##
## locale:
## [1] LC_CTYPE=en_US.UTF-8 LC_NUMERIC=C
## [3] LC_TIME=en_GB LC_COLLATE=C
## [5] LC_MONETARY=en_US.UTF-8 LC_MESSAGES=en_US.UTF-8
## [7] LC_PAPER=en_US.UTF-8 LC_NAME=C
## [9] LC_ADDRESS=C LC_TELEPHONE=C
## [11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C
##
## time zone: America/New_York
## tzcode source: system (glibc)
##
## attached base packages:
## [1] stats4 stats graphics grDevices utils datasets methods
## [8] base
##
## other attached packages:
## [1] tidySingleCellExperiment_1.10.0 ttservice_0.2.2
## [3] tidyHeatmap_1.8.1 purrr_1.0.1
## [5] SingleCellSignalR_1.12.0 SingleR_2.2.0
## [7] scran_1.28.0 scater_1.28.0
## [9] ggplot2_3.4.2 scuttle_1.10.0
## [11] SingleCellExperiment_1.22.0 SummarizedExperiment_1.30.0
## [13] Biobase_2.60.0 GenomicRanges_1.52.0
## [15] GenomeInfoDb_1.36.0 IRanges_2.34.0
## [17] S4Vectors_0.38.0 BiocGenerics_0.46.0
## [19] MatrixGenerics_1.12.0 matrixStats_0.63.0
## [21] knitr_1.42
##
## loaded via a namespace (and not attached):
## [1] RColorBrewer_1.1-3 jsonlite_1.8.4
## [3] shape_1.4.6 magrittr_2.0.3
## [5] magick_2.7.4 ggbeeswarm_0.7.1
## [7] farver_2.1.1 GlobalOptions_0.1.2
## [9] zlibbioc_1.46.0 vctrs_0.6.2
## [11] multtest_2.56.0 Cairo_1.6-0
## [13] DelayedMatrixStats_1.22.0 RCurl_1.98-1.12
## [15] htmltools_0.5.5 BiocNeighbors_1.18.0
## [17] KernSmooth_2.23-20 htmlwidgets_1.6.2
## [19] plyr_1.8.8 plotly_4.10.1
## [21] igraph_1.4.2 lifecycle_1.0.3
## [23] iterators_1.0.14 pkgconfig_2.0.3
## [25] rsvd_1.0.5 Matrix_1.5-4
## [27] R6_2.5.1 fastmap_1.1.1
## [29] GenomeInfoDbData_1.2.10 clue_0.3-64
## [31] digest_0.6.31 reshape_0.8.9
## [33] GGally_2.1.2 colorspace_2.1-0
## [35] patchwork_1.1.2 dqrng_0.3.0
## [37] irlba_2.3.5.1 beachmat_2.16.0
## [39] labeling_0.4.2 fansi_1.0.4
## [41] httr_1.4.5 compiler_4.3.0
## [43] withr_2.5.0 doParallel_1.0.17
## [45] BiocParallel_1.34.0 viridis_0.6.2
## [47] highr_0.10 dendextend_1.17.1
## [49] gplots_3.1.3 MASS_7.3-59
## [51] DelayedArray_0.26.0 rjson_0.2.21
## [53] bluster_1.10.0 gtools_3.9.4
## [55] caTools_1.18.2 tools_4.3.0
## [57] vipor_0.4.5 beeswarm_0.4.0
## [59] glue_1.6.2 grid_4.3.0
## [61] Rtsne_0.16 cluster_2.1.4
## [63] generics_0.1.3 gtable_0.3.3
## [65] tidyr_1.3.0 data.table_1.14.8
## [67] BiocSingular_1.16.0 ScaledMatrix_1.8.0
## [69] metapod_1.8.0 utf8_1.2.3
## [71] XVector_0.40.0 ggrepel_0.9.3
## [73] foreach_1.5.2 pillar_1.9.0
## [75] stringr_1.5.0 limma_3.56.0
## [77] circlize_0.4.15 splines_4.3.0
## [79] dplyr_1.1.2 lattice_0.21-8
## [81] FNN_1.1.3.2 survival_3.5-5
## [83] tidyselect_1.2.0 ComplexHeatmap_2.16.0
## [85] locfit_1.5-9.7 gridExtra_2.3
## [87] edgeR_3.42.0 xfun_0.39
## [89] dittoSeq_1.12.0 statmod_1.5.0
## [91] pheatmap_1.0.12 stringi_1.7.12
## [93] lazyeval_0.2.2 evaluate_0.20
## [95] codetools_0.2-19 tibble_3.2.1
## [97] BiocManager_1.30.20 cli_3.6.1
## [99] uwot_0.1.14 munsell_0.5.0
## [101] Rcpp_1.0.10 png_0.1-8
## [103] parallel_4.3.0 ellipsis_0.3.2
## [105] sparseMatrixStats_1.12.0 bitops_1.0-7
## [107] viridisLite_0.4.1 ggridges_0.5.4
## [109] scales_1.2.1 crayon_1.5.2
## [111] GetoptLong_1.0.5 rlang_1.1.0
## [113] cowplot_1.1.1