Estimate spot-level cell-type proportions from spatial expression and an annotated single-cell reference using CARD or CARDspa.
Usage
RunCARD(
object,
reference,
reference_label,
assay = NULL,
reference_assay = NULL,
layer = "counts",
reference_layer = "counts",
features = NULL,
image = NULL,
coord.cols = c("col", "row"),
sample_varname = NULL,
minCountGene = 100,
minCountSpot = 5,
ct_select = NULL,
prefix = "CARD",
tool_name = "CARD",
store_results = TRUE,
round_counts = TRUE,
create_card_params = list(),
card_deconvolution_params = list(),
verbose = TRUE,
...,
coordinate_space = c("raw", "legacy_display"),
srt = NULL
)Arguments
- object
Spatial
Seuratobject used as the RCTD query.- reference
Reference
Seuratobject containing annotated single cells.- reference_label
Metadata column in
referencewith cell type labels.- assay
Assay used in
object. IfNULL, the default assay is used.- reference_assay
Assay used in
reference. Only cells present in this assay are used, with reference labels aligned by cell ID.- layer, reference_layer
Assay layers used for spatial and reference raw counts.
- features
Features used for CARD. If
NULL, shared features are used.- image
Name of the Seurat spatial image used to recover coordinates when
coord.colsare not available.- coord.cols
Metadata coordinate columns used when no image coordinate source is requested or available.
- sample_varname
Optional metadata column in
referencecontaining sample labels. WhenNULL, all reference cells are assigned to one sample.- minCountGene, minCountSpot
Filtering parameters passed to
CARD::createCARDObject()orCARDspa::createCARDObject()when supported.- ct_select
Optional cell types to keep in CARD.
- prefix
Prefix for metadata columns.
- tool_name
Name used to store detailed results in
srt@tools.- store_results
Whether to store detailed RCTD results in
srt@tools.- round_counts
Whether to round non-integer values before passing data to
spacexr. RCTD requires integer count matrices; rounding is a compatibility option and does not recover original counts.- create_card_params
Additional parameters passed to
createCARDObject().- card_deconvolution_params
Additional parameters passed to
CARD_deconvolution().- verbose
Whether to print the message. Default is
TRUE.- ...
Additional parameters passed to the RCTD run step.
- coordinate_space
Coordinate space used for distance-sensitive input. The default is raw acquisition coordinates, so backend distances use raw coordinate units. Use
"legacy_display"explicitly to reproduce the display-scaled coordinates used before scop 0.9.0.- srt
Deprecated alias for
object; supply exactly one of the two. It will be removed in scop 1.0.0.
Value
A Seurat object with CARD proportion columns in metadata and
dominant cell type summaries. When store_results = TRUE, detailed results
are stored in srt@tools[[tool_name]].
Examples
if (FALSE) { # \dontrun{
data(visium_human_pancreas_sub)
data(panc8_sub)
keep_spots <- unique(round(seq(
1,
ncol(visium_human_pancreas_sub),
length.out = 120
)))
spatial <- visium_human_pancreas_sub[, keep_spots]
# Results are conditional on the three reference cell types used here.
reference <- panc8_sub[, panc8_sub@meta.data[["celltype"]] %in%
c("ductal", "alpha", "beta")]
features_use <- head(intersect(rownames(spatial), rownames(reference)), 300)
spatial <- RunCARD(
object = spatial,
reference = reference,
reference_label = "celltype",
assay = "Spatial",
reference_assay = "RNA",
layer = "counts",
reference_layer = "counts",
features = features_use,
coord.cols = c("x", "y"),
verbose = FALSE
)
SpatialDeconvolutionPlot(
spatial,
tool_name = "CARD",
plot_type = "dominant",
overlay_image = FALSE,
coord.cols = c("x", "y")
)
} # }