plotCells {cytomapper} | R Documentation

## Function to visualize cell-level information on segmentation masks

### Description

This function takes a `SingleCellExperiment` and `CytoImageList` object containing segmentation masks to colour cells by marker expression or metadata.

### Usage

    plotCells(
      mask,
      object = NULL,
      cell_id = NULL,
      img_id = NULL,
      colour_by = NULL,
      outline_by = NULL,
      exprs_values = "counts",
      colour = NULL,
      ...
    )

### Arguments

`mask` | a `CytoImageList` containing single-channel `Image` objects (see section 'Segmentation mask object' below).
`object` | a `SingleCellExperiment` object.
`cell_id` | character specifying the `colData(object)` entry, in which the integer cell IDs are stored. These IDs should match the integer pixel values in the segmentation mask object.
`img_id` | character specifying the `colData(object)` and `mcols(mask)` entry, in which the image IDs are stored (see section 'Linking the `SingleCellExperiment` and `CytoImageList` object' below)
`colour_by` | character or character vector specifying the features (`rownames(object)`) or metadata (`colData(object)` entry) used to colour individual cells. Cells can be coloured by single `colData(object)` entries or by up to six features.
`outline_by` | single character indicating the `colData(object)` entry by which to outline individual cells.
`exprs_values` | single character indicating which `assay(object)` entry to use when visualizing feature counts.
`colour` | a list with names matching the entries to `colour_by` and/or `outline_by`. When setting the colour for continous features, at least two colours need to be provided indicating the colours for minimum and maximum values. When colouring discrete vectors, a colour for each unique entry needs to be provided (see section 'Setting the colours' and examples)
`...` | Further arguments passed to `?"plotting-param"`

### Value

a list if `return_images` and/or `return_plot` is TRUE (see `?"plotting-param"`).

- `plot`: a single plot object (`display = "all"`) or a list of plot objects (`display = "single"`)

- `images`: a `SimpleList` object containing three-colour `Image` objects.

### Segmentation mask object

In the `plotCells` function, `mask` refers to a `CytoImageList` object that contains a single or multiple segmentation masks in form of individual `Image` objects. The function assumes that each object in the segmentation mask is a cell. The key features of such masks include:

- each Image object contains only one channel

- pixel values are integers indicating the cells' IDs or 0 (background)

### Linking SingleCellExperiment and CytoImageList objects

To colour individual cells contained in the segmentation masks based on features and metadata stored in the SingleCellExperiment object, an `img_id` and `cell_id` entry needs to be provided. Image IDs are matched between the `SingleCellExperiment` and `CytoImageList` object via entries to the `colData(object)[,img_id]` and the `mcols(mask)[,img_id]` slots. Cell IDs are matched between the `SingleCellExperiment` and `CytoImageList` object via entries to `colData(object)[,cell_id]` and the integer values of the segmentation masks.

### Setting the colours

By default, features and metadata are coloured based on internally-set colours. To set new colours, a `list` object must be provided. The names of the object must correspond to the entries to `colour_by` and/or `outline_by`. When setting the colours for continous expression values or continous metadata entries, a vector of at least two colours need to be specified. These colours will be passed onto `colorRampPalette` for interpolation. Discrete metadata entries can be coloured by specifying a named vector in which each entry corresponds to a unique entry to the metadata vector.

### Subsetting the `CytoImageList` object

The `CytoImageList` object can be subsetted before calling the `plotCells` function. In that case, only the selected images are displayed.

### Subsetting the `SingleCellExperiment` object

The `SingleCellExperiment` object can be subsetted before calling the `plotCells` function. In that case, only cells contained in the `SingleCellExperiment` object are coloured/outlined.

### Colour scaling

When colouring features using the plotCells function, colours are scaled between the minimum and maximum per feature across the full assay contained in the SingleCellExperiment object. When subsetting images, cell-level expression is not scaled across the subsetted images but the whole SingleCellExperiment object. To avoid this, the SingleCellExperiment object can be subsetted to only contain the cells that should be displayed before plotting.

### Author(s)

Nils Eling (nils.eling@dqbm.uzh.ch)

Nicolas Damond (nicolas.damond@dqbm.uzh.ch)

### See Also

For further plotting parameters see `?"plotting-param"`

### Examples

    data(pancreasMasks)
    data(pancreasSCE)

    # Visualize the masks
    plotCells(pancreasMasks)

    # Colour the masks based on averaged expression
    plotCells(pancreasMasks, object = pancreasSCE, img_id = "ImageNb",
                cell_id = "CellNb", colour_by = c("CD99", "CDH"))

    # Colour the masks based on metadata
    plotCells(pancreasMasks, object = pancreasSCE, img_id = "ImageNb",
                cell_id = "CellNb", colour_by = "CellType")

    # Outline the masks based on metadata
    plotCells(pancreasMasks, object = pancreasSCE, img_id = "ImageNb",
                cell_id = "CellNb", colour_by = "CD99",
                outline_by = "CellType")

    # Colour the masks based on arcsinh-transformed expression
    plotCells(pancreasMasks, object = pancreasSCE, img_id = "ImageNb",
                cell_id = "CellNb", colour_by = "CD99",
                exprs_values = "exprs")

    # Subset the images
    cur_images <- getImages(pancreasMasks, 1:2)
    plotCells(cur_images, object = pancreasSCE, img_id = "ImageNb",
                cell_id = "CellNb", colour_by = "CD99")

    # Set colour
    plotCells(pancreasMasks, object = pancreasSCE, img_id = "ImageNb",
                cell_id = "CellNb", colour_by = "CD99", outline_by = "CellType",
                colour = list(CD99 = c("black", "red"),
                                CellType = c(celltype_A = "blue",
                                            celltype_B = "green",
                                            celltype_C = "red")))

[Package cytomapper version 1.4.1 Index]