This vignette consists of a series of (independent) hands-on recipes, aimed at exploring the capabilities of iSEE both interactively and programmatically. Each recipe consists of a short task and a screen shot of the desired appearance of the app instance that should be created. For each recipe, we provide a set of hints, as well as detailed instructions on how to solve the task both interactively (by clicking in the app) and programmatically (by directly setting up and launching the desired app instance).
For a general overview of the default iSEE panels,
we refer to the overview vignette. For all the details about the panel
classes and the associated slots, we refer to the help pages for the
respective panel class (e.g., ?ReducedDimensionPlot).
Before starting with the recipes, we need to load the required packages and the demo data set (PBMC3k). You can download the data set from Dropbox.
Using the PBMC3k data set, create an app that contains only a reduced dimension plot panel, a row data plot panel and a column data plot panel. The widths of the three panels should be 3, 4 and 5 units, respectively.
Organization button in the top right corner of the app.
initial argument to iSEE().
Organization button in the top right corner of
the app, and then click on Organize panels.
x
next to all the panels that you want to remove (all but the
Reduced dimension plot 1, Column data plot 1
and Row data plot 1).
Width dropdown menu.
Apply settings.
Using the PBMC3k data set, visualize the cell type assignment against the cluster membership, with the aim to identify the predominant cell type in each cluster. In this case, since both cell annotations are categorical, iSEE will generate a so called Hinton plot.
ColumnDataPlot panel.
Cluster column of
colData(sce).
labels_fine
column of colData(sce) (more coarse-grained assignments are
provided in the labels_main column).
ColumnDataPlot
panel spanning the full application window)
ColumnDataPlot panel, click to expand the
Data parameters collapsible box.
Column of interest (Y-axis), select the label column
(e.g., labels_fine).
X-axis, select Column data, and under
Column of interest (X-axis), select Cluster.
Using the PBMC3k data set, display both the tSNE and UMAP representations next to each other.
ReducedDimensionPlot panels.
reducedDimNames(sce) lists the
available representations).
ReducedDimensionPlot
panels, each spanning half the application window)
app <- iSEE(sce, initial = list(ReducedDimensionPlot(PanelWidth = 6L),
ReducedDimensionPlot(PanelWidth = 6L)))
shiny::runApp(app)ReducedDimensionPlot panel, click to expand
the Data parameters collapsible box.
Type selection box, choose TSNE.
ReducedDimensionPlot panel, repeat the
procedure but instead select UMAP.
Using the PBMC3k data set, plot the distribution of the
logcount values for the gene CD74 in each of the
clusters.
FeatureAssayPlot panel.
FeatureAssayPlot panel,
spanning the full application window)
FeatureAssayPlot panel, click to expand the
Data parameters collapsible box.
Y-axis feature, type or select CD74.
X-axis, select Column data, and under
Column of interest (X-axis), select Cluster.
Using the PBMC3k data set, display two tSNE representations
next to each other. In the first one, color the cells by the cluster
label. In the second one, color the cells by the total UMI count
(total column of colData(sce)).
ReducedDimensionPlot panels.
reducedDimNames(sce) lists the
available representations).
Cluster column of
colData(sce).
Visual parameters
collapsible box. To display or hide the possible options, check the
corresponding checkboxes (Color, Shape,
Size, Point).
ReducedDimensionPlot
panels, each spanning half the application window)
app <- iSEE(sce, initial = list(ReducedDimensionPlot(PanelWidth = 6L),
ReducedDimensionPlot(PanelWidth = 6L)))
shiny::runApp(app)ReducedDimensionPlot panel, click to expand the
Data parameters collapsible box and under
Type, choose TSNE.
ReducedDimensionPlot panel, click to expand
the Visual parameters collapsible box.
Color checkbox is ticked.
Color by, select Column data.
Cluster.
ReducedDimensionPlot panel, repeat the
procedure but instead select total.
Using the PBMC3k data set, display two tSNE representations
next to each other. In the first one, color the cells by the logcounts
expression level of CD3D. In the second one, color the
cells by the logcounts expression level of CD79B.
ReducedDimensionPlot panels.
reducedDimNames(sce) lists the
available representations).
Visual parameters
collapsible box. To display or hide the possible options, check the
corresponding checkboxes (Color, Shape,
Size, Point).
ReducedDimensionPlot
panels, each spanning half the application window)
app <- iSEE(sce, initial = list(ReducedDimensionPlot(PanelWidth = 6L),
ReducedDimensionPlot(PanelWidth = 6L)))
shiny::runApp(app)ReducedDimensionPlot panel, click to expand the
Data parameters collapsible box and under
Type, choose TSNE.
ReducedDimensionPlot panel, click to expand
the Visual parameters collapsible box.
Color checkbox is ticked.
Color by, select Feature name.
CD3D.
ReducedDimensionPlot panel, repeat the
procedure but instead select CD79B.
Using the PBMC3k data set, display two tSNE representations next to each other. In the first one, set the point size to 0.5. In the second one, set the point size to 3 and the opacity to 0.2.
ReducedDimensionPlot panels.
reducedDimNames(sce) lists the
available representations).
Visual parameters
collapsible box. To display or hide the possible options, check the
corresponding checkboxes (Color, Shape,
Size, Point).
ReducedDimensionPlot
panels, each spanning half the application window)
app <- iSEE(sce, initial = list(ReducedDimensionPlot(PanelWidth = 6L),
ReducedDimensionPlot(PanelWidth = 6L)))
shiny::runApp(app)ReducedDimensionPlot panel, click to expand the
Data parameters collapsible box and under
Type, choose TSNE.
ReducedDimensionPlot panel, click to expand
the Visual parameters collapsible box.
Size checkbox is ticked.
Size by, select None.
ReducedDimensionPlot panel, click to expand
the Visual parameters collapsible box.
Size and Point checkboxes
are ticked.
Size by, select None.
Point opacity, drag the slider to 0.2.
Using the PBMC3k data set, display the expression
(logcounts) of CD3D across the assigned clusters, as well
as a tSNE representation colored by the cluster label. Select all cells
with a logcount expression value of CD3D between
(approximately) 0.5 and 4, and highlight these in the tSNE plot by means
of transparency.
ReducedDimensionPlot panels.
reducedDimNames(sce) lists the
available representations).
FeatureAssayPlot panel.
Selection parameters collapsible box.
FeatureAssayPlot panel
and one ReducedDimensionPlot panel, each spanning half the
application window)
app <- iSEE(sce, initial = list(FeatureAssayPlot(PanelWidth = 6L),
ReducedDimensionPlot(PanelWidth = 6L)))
shiny::runApp(app)FeatureAssayPlot panel, click to expand the
Data parameters collapsible box and under
Y-axis feature, type or select CD3D.
X-axis, select Column data, and under
X-axis column data, select Cluster.
FeatureAssayPlot panel, use the mouse to drag a
rectangle around all points with a logcount expression value (y-axis)
between approximately 0.5 and 4.
ReducedDimensionPlot panel, click to expand the
Data parameters collapsible box and under
Type, choose TSNE.
ReducedDimensionPlot panel, further click to expand
the Visual parameters collapsible box, make sure that the
Color checkbox is ticked. Under Color by,
select Column data. In the dropdown menu that appears, type
or select Cluster.
ReducedDimensionPlot panel, click to expand the
Selection parameters collapsible box.
Receive column selection from, select
Feature assay plot 1.
app <- iSEE(sce, initial = list(
FeatureAssayPlot(PanelWidth = 6L,
BrushData = list(xmin = 0, xmax = 15,
ymin = 0.5, ymax = 4,
mapping = list(x = "X", y = "Y", group = "GroupBy"),
direction = "xy",
brushId = "FeatureAssayPlot1_Brush",
outputId = "FeatureAssayPlot1"),
XAxis = "Column data",
XAxisColumnData = "Cluster", YAxisFeatureName = "CD3D"),
ReducedDimensionPlot(PanelWidth = 6L, Type = "TSNE",
ColorBy = "Column data",
ColorByColumnData = "Cluster",
ColumnSelectionSource = "FeatureAssayPlot1")
))Using the PBMC3k data set, create a scatter plots displaying
the (logcounts) expression values of CD79A vs
CD74, as well as a Hinton plot of the cluster and cell type
assignment annotations. Select the cells co-expressing
CD79A and CD74 in the scatter plot. Which cell
type/cluster(s) do these correspond to (color these points in the Hinton
plot)?
FeatureAssayPlot panel.
ColumnDataPlot panel.
Cluster column of
colData(sce). Coarse-grained cell type labels are available
in the labels_main column.
Selection parameters collapsible box.
FeatureAssayPlot panel
and one ColumnDataPlot panel, each spanning half of the
application window)
app <- iSEE(sce, initial = list(FeatureAssayPlot(PanelWidth = 6L),
ColumnDataPlot(PanelWidth = 6L)))
shiny::runApp(app)FeatureAssayPlot panel, click to expand the
Data parameters collapsible box.
Y-axis feature, type or select CD79A.
X-axis, select Feature name, and under
X-axis feature, type or select CD74.
ColumnDataPlot panel, click to expand the
Data parameters collapsible box.
Column of interest (Y-axis), type or select
labels_main.
X-axis, select Column data, and under
Column of interest (X-axis), select Cluster.
FeatureAssayPlot panel, use the mouse to drag a
rectangle around all points co-expressing CD79A and
CD74.
ColumnDataPlot panel, click to expand the
Selection parameters collapsible box.
Receive column selection from, select
Feature assay plot 1.
ColumnDataPlot panel, click to expand the
Visual parameters collapsible box.
Color by, select Column selection.
app <- iSEE(sce, initial = list(
FeatureAssayPlot(PanelWidth = 6L, XAxis = "Feature name",
YAxisFeatureName = "CD79A",
XAxisFeatureName = "CD74",
BrushData = list(
xmin = 0.3, xmax = 7,
ymin = 0.3, ymax = 7,
mapping = list(x = "X", y = "Y", colour = "ColorBy"),
direction = "xy", brushId = "FeatureAssayPlot1_Brush",
outputId = "FeatureAssayPlot1")),
ColumnDataPlot(PanelWidth = 6L, XAxis = "Column data",
YAxis = "labels_main",
XAxisColumnData = "Cluster",
ColumnSelectionSource = "FeatureAssayPlot1",
ColorBy = "Column selection")
))Using the PBMC3k data set, load iSEEu and
use the modeReducedDim mode to open an app displaying all
the reduced dimension representations stored in the SingleCellExperiment
object. Color the representations by the cell type assignment.
labels_fine
column of colData(sce) (more coarse-grained assignments are
provided in the labels_main column).
iSEEu::modeReducedDim().
Using the PBMC3k data set, display two tSNE representations
next to each other. In the first one, color the cells by the logcounts
expression level of CD3D. In the second one, color the
cells by the logcounts expression level of CD79B. Also
include a small tour that starts with a welcome message, next walks
through the two panels, giving an informative message for each, and
finally ends with a concluding message to the user.
ReducedDimensionPlot panels.
reducedDimNames(sce) lists the
available representations).
Visual parameters
collapsible box. To display or hide the possible options, check the
corresponding checkboxes (Color, Shape,
Size, Point).
tour argument to
iSEE().
element and intro. The element
column contains the names of UI elements, prefixed by a hash sign. More
details, including how to find the name of a particular UI elements, can
be found in the Configuring iSEE apps
vignette of iSEE.
tour <- data.frame(
element = c(
"#Welcome",
"#ReducedDimensionPlot1",
"#ReducedDimensionPlot2",
"#Conclusion"),
intro = c(
"Welcome to this tour!",
"This is the first reduced dimension plot",
"And here is the second one",
"Thank you for taking this tour!"),
stringsAsFactors = FALSE)
app <- iSEE(sce, initial = list(
ReducedDimensionPlot(PanelWidth = 6L, Type = "TSNE",
ColorBy = "Feature name",
ColorByFeatureName = "CD3D"),
ReducedDimensionPlot(PanelWidth = 6L, Type = "TSNE",
ColorBy = "Feature name",
ColorByFeatureName = "CD79B")),
tour = tour)The main focus of iSEE is
exploration of pre-calculated results. This is reflected in the default
set of panels, which all rely on reading and displaying information
stored in the SummarizedExperiment object. However, it is
also possible to construct custom panels, which can perform arbitrarily
complex analyses. Providing a middle ground, the iSEEu
package contains some custom panel types that can perform basic
analysis, such as a simple test for differential expression between
selected groups of samples. It is worth noting that although there is
not a single, ‘always-optimal’ way of doing this type of analysis, the
panel type provides a quick way of characterizing a set of samples via
the set of upregulated genes.
Using the PBMC3k data set, display a tSNE representation colored by the cluster assignment, as well as a dynamic marker table that displays genes that are differentially expressed between sets of interactively selected cells in the tSNE plot. Select the smallest cluster in the bottom of the plot and find the genes most strongly upregulated compared to the other cells.
ReducedDimensionPlot panels.
reducedDimNames(sce) lists the
available representations).
DynamicMarkerTable panel).
iSEE()) you need to provide the panel class to the
extra argument of iSEE(), unless you specify
the panel in the initial argument.
ReducedDimensionPlot
panel and one DynamicMarkerTable panel, each spanning half
of the application window). Note that in order to access the
DynamicMarkerTable panel type, the iSEEu
package must be loaded into your R session.
library(iSEEu)
app <- iSEE(sce, initial = list(ReducedDimensionPlot(PanelWidth = 6L),
DynamicMarkerTable(PanelWidth = 6L)))
shiny::runApp(app)Organize panels button, you can launch iSEE with
the default set of panels. In this case, make sure to provide the
DynamicMarkerTable to the extra argument, so
that it will show up as an option when you add new panels. If you take
this approach, click on the Organization button in the top
right corner of the app, and then click on Organize panels
and set up the desired interface.
ReducedDimensionPlot panel, click to expand the
Data parameters collapsible box.
Type, type or select TSNE.
Visual parameters collapsible box.
Color checkbox is ticked.
Color by, select Column data.
Cluster.
DynamicMarkerTable panel, click to expand the
Selection parameters collapsible box.
Receive column selection from, select
Reduced dimension plot 1.
app <- iSEE(sce, initial = list(
ReducedDimensionPlot(PanelWidth = 6L, Type = "TSNE",
ColorBy = "Column data",
ColorByColumnData = "Cluster",
BrushData = list(xmin = -20, xmax = 5,
ymin = -40, ymax = -15,
mapping = list(x = "X", y = "Y",
colour = "ColorBy"),
direction = "xy",
brushId = "ReducedDimensionPlot1_Brush",
outputId = "ReducedDimensionPlot1")),
DynamicMarkerTable(PanelWidth = 6L,
ColumnSelectionSource = "ReducedDimensionPlot1"))
)sessionInfo()
#> R version 4.4.0 (2024-04-24)
#> Platform: aarch64-apple-darwin20
#> Running under: macOS Sonoma 14.5
#>
#> Matrix products: default
#> BLAS: /Library/Frameworks/R.framework/Versions/4.4-arm64/Resources/lib/libRblas.0.dylib
#> LAPACK: /Library/Frameworks/R.framework/Versions/4.4-arm64/Resources/lib/libRlapack.dylib; LAPACK version 3.12.0
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#>
#> time zone: Europe/Zurich
#> tzcode source: internal
#>
#> attached base packages:
#> [1] stats4 stats graphics grDevices utils datasets methods
#> [8] base
#>
#> other attached packages:
#> [1] iSEEu_1.16.0 iSEEhex_1.6.0
#> [3] iSEE_2.16.0 SingleCellExperiment_1.26.0
#> [5] SummarizedExperiment_1.34.0 Biobase_2.64.0
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