Mercurial > repos > iuc > scater_plot_tsne
annotate scater-plot-dist-scatter.R @ 2:99f912d5af9f draft default tip
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 9961b5acbf9081f10e14bc272406b36854fa2924"
author | iuc |
---|---|
date | Mon, 08 Nov 2021 12:03:09 +0000 |
parents | 2b09ca1c5e41 |
children |
rev | line source |
---|---|
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
1 #!/usr/bin/env Rscript |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
2 |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
3 # Plot the distribution of read counts and feature counts, side by side, then a scatter plot of read counts vs feature counts below |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
4 |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
5 # Load optparse we need to check inputs |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
6 |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
7 library(optparse) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
8 library(workflowscriptscommon) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
9 library(LoomExperiment) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
10 library(scater) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
11 library(ggpubr) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
12 library(scales) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
13 |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
14 # parse options |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
15 |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
16 option_list <- list( |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
17 make_option( |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
18 c("-i", "--input-loom"), |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
19 action = "store", |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
20 default = NA, |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
21 type = "character", |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
22 help = "A SingleCellExperiment object file in Loom format." |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
23 ), |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
24 make_option( |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
25 c("-o", "--output-plot-file"), |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
26 action = "store", |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
27 default = NA, |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
28 type = "character", |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
29 help = "Path of the PDF output file to save plot to." |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
30 ), |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
31 make_option( |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
32 c("-l", "--log-scale"), |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
33 action = "store_true", |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
34 default = FALSE, |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
35 type = "logical", |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
36 help = "Plot on log scale (recommended for large datasets)." |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
37 ) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
38 ) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
39 |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
40 opt <- wsc_parse_args(option_list, mandatory = c("input_loom", "output_plot_file")) |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
41 |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
42 # Check parameter values |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
43 |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
44 if (! file.exists(opt$input_loom)) { |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
45 stop((paste("File", opt$input_loom, "does not exist"))) |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
46 } |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
47 |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
48 # Filter out unexpressed features |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
49 |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
50 sce <- import(opt$input_loom, format = "loom", type = "SingleCellLoomExperiment") |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
51 |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
52 # Do the scatter plot of reads vs genes |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
53 total_counts <- sce$total |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
54 total_features <- sce$detected |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
55 count_feats <- cbind(total_counts, total_features) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
56 cf_dm <- as.data.frame(count_feats) |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
57 |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
58 # Calculate binwidths for reads and features plots. Use 20 bins |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
59 read_bins <- max(total_counts / 1e6) / 20 |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
60 feat_bins <- max(total_features) / 20 |
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
61 |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
62 plot1 <- qplot(total_counts / 1e6, geom = "histogram", binwidth = read_bins, ylab = "Number of cells", xlab = "Read counts (millions)", fill = I("darkseagreen3")) + ggtitle("Read counts per cell") |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
63 plot2 <- qplot(total_features, geom = "histogram", binwidth = feat_bins, ylab = "Number of cells", xlab = "Feature counts", fill = I("darkseagreen3")) + ggtitle("Feature counts per cell") |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
64 plot3 <- ggplot(cf_dm, aes(x = total_counts / 1e6, y = total_features)) + geom_point(shape = 1) + geom_smooth() + xlab("Read count (millions)") + |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
65 ylab("Feature count") + ggtitle("Scatterplot of reads vs features") |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
66 plot4 <- plotColData(sce, y = "subsets_Mito_percent", x = "detected") + ggtitle("% MT genes") + geom_point(shape = 1) + theme(text = element_text(size = 15)) + theme(plot.title = element_text(size = 15)) + xlab("Total features") + ylab("% MT") |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
67 |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
68 if (! opt$log_scale) { |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
69 final_plot <- ggarrange(plot1, plot2, plot3, plot4, ncol = 2, nrow = 2) |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
70 ggsave(opt$output_plot_file, final_plot, device = "pdf") |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
71 } else { |
1
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
72 plot1_log <- plot1 + scale_x_continuous(trans = "log10") + scale_y_continuous(trans = "log10") |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
73 plot2_log <- plot2 + scale_y_continuous(trans = "log10") |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
74 plot3_log <- plot3 + scale_y_continuous(trans = "log10") |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
75 plot4_log <- plot4 + scale_y_log10(labels = number) |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
76 final_plot_log <- ggarrange(plot1_log, plot2_log, plot3_log, plot4_log, ncol = 2, nrow = 2) |
2b09ca1c5e41
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 154318f74839a4481c7c68993c4fb745842c4cce"
iuc
parents:
0
diff
changeset
|
77 ggsave(opt$output_plot_file, final_plot_log, device = "pdf") |
0
a30f4bfe8f01
"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/scater commit 61f3899168453092fd25691cf31871a3a350fd3b"
iuc
parents:
diff
changeset
|
78 } |