Add transformed array and pixel coordinates to a SpatialExperiment

Given a SpatialExperiment-class, sample information, and coordinates produced from the refinement workflow, add array and pixel coordinates appropriate for the linearly transformed capture areas making up each group present in the SpatialExperiment-class.

Usage

add_array_coords(spe, sample_info, coords_dir, calc_error_metrics = FALSE)

Arguments

spe: A SpatialExperiment-class object.
sample_info: A data.frame() with columns capture_area, group, fiji_xml_path, fiji_image_path, spaceranger_dir, intra_group_scalar, and group_hires_scalef. The last two are made by rescale_fiji_inputs().
coords_dir: A character(1) vector giving the directory containing sample directories each with tissue_positions.csv, scalefactors_json.json, and tissue_lowres_image.png files produced from refinement with prep_fiji_coords() and related functions.
calc_error_metrics: A logical(1) vector indicating whether to calculate error metrics related to mapping spots to well-defined array coordinates. If TRUE, adds euclidean_error and shared_neighbors spot-level metrics to the colData(). The former indicates distance in number of inter-spot distances to "move" a spot to the new array position; the latter indicates the fraction of neighbors for the associated capture area that are retained after mapping, which can be quite time-consuming to compute.

Value

A SpatialExperiment-class object with additional colData columns pxl_row_in_fullres_[suffix] and pxl_col_in_fullres_[suffix] with [suffix] values original and rounded; array_row_original and array_col_original columns; and modified colData() columns array_row and array_col and spatialCoords() with their transformed values.

Details

Array coordinates are determined via an algorithm that fits each spot to the nearest spot on a new, imaginary, Visium-like capture area. The imaginary capture area differs from a real capture area only in its extent; array coordinates still start at 0 but may extend arbitrarily beyond the normal maximum indices of 77 and 127 to fit every capture area in each group defined in the SpatialExperiment-class. The goal is to return well-defined array coordinates in a consistent spatial orientation for each group, such that downstream applications, such as clustering with BayesSpace, can process each group as if it really were one capture area in the first place. See https://research.libd.org/visiumStitched/articles/visiumStitched.html#defining-array-coordinates for more details.

Author

Nicholas J. Eagles

Examples

if (!exists("spe")) {
    spe <- spatialLIBD::fetch_data(type = "visiumStitched_brain_spe")
}
#> 2024-11-08 18:52:00.791712 loading file /github/home/.cache/R/BiocFileCache/6c1f1cbefd45_visiumStitched_brain_spe.rds%3Frlkey%3Dnq6a82u23xuu9hohr86oodwdi%26dl%3D1

########################################################################
#   Prepare sample_info
########################################################################

sample_info <- dplyr::tibble(
    group = "Br2719",
    capture_area = c("V13B23-283_A1", "V13B23-283_C1", "V13B23-283_D1")
)
#   Add 'spaceranger_dir' column
sr_dir <- tempdir()
temp <- unzip(
    spatialLIBD::fetch_data("visiumStitched_brain_spaceranger"),
    exdir = sr_dir
)
#> 2024-11-08 18:52:05.968245 loading file /github/home/.cache/R/BiocFileCache/6c1f5dcc09d3_visiumStitched_brain_spaceranger.zip%3Frlkey%3Dbdgjc6mgy1ierdad6h6v5g29c%26dl%3D1
sample_info$spaceranger_dir <- file.path(
    sr_dir, sample_info$capture_area, "outs", "spatial"
)

#   Add Fiji-output-related columns
fiji_dir <- tempdir()
temp <- unzip(
    spatialLIBD::fetch_data("visiumStitched_brain_Fiji_out"),
    exdir = fiji_dir
)
#> 2024-11-08 18:52:08.610431 loading file /github/home/.cache/R/BiocFileCache/6c1f40e8b6a3_visiumStitched_brain_fiji_out.zip%3Frlkey%3Dptwal8f5zxakzejwd0oqw0lhj%26dl%3D1
sample_info$fiji_xml_path <- temp[grep("xml$", temp)]
sample_info$fiji_image_path <- temp[grep("png$", temp)]

## Re-size images and add more information to the sample_info
sample_info <- rescale_fiji_inputs(sample_info, out_dir = tempdir())

## Preparing Fiji coordinates and images for build_SpatialExperiment()
spe_input_dir <- tempdir()
prep_fiji_coords(sample_info, out_dir = spe_input_dir)
#> [1] "/tmp/RtmpYqIEhJ/Br2719/tissue_positions.csv"
prep_fiji_image(sample_info, out_dir = spe_input_dir)
#> [1] "/tmp/RtmpYqIEhJ/Br2719/tissue_lowres_image.png"
#> [2] "/tmp/RtmpYqIEhJ/Br2719/scalefactors_json.json" 

########################################################################
#   Add array coordinates
########################################################################

spe_new <- add_array_coords(spe, sample_info, tempdir())

#    Several columns related to spatial coordinates were added
added_cols_regex <- "^(array|pxl)_(row|col)(_in_fullres)?_(original|rounded)$"
colnames(SummarizedExperiment::colData(spe_new))[
    grep(added_cols_regex, colnames(SummarizedExperiment::colData(spe_new)))
]
#> [1] "array_row_original"          "array_col_original"         
#> [3] "pxl_col_in_fullres_rounded"  "pxl_row_in_fullres_rounded" 
#> [5] "pxl_row_in_fullres_original" "pxl_col_in_fullres_original"

#    'array_row', 'array_col', and spatialCoords() were overwritten with
#    their transformed values
head(spe$array_row)
#> [1]  0 50  3 59 14 43
head(spe$array_col)
#> [1]  16 102  43  19  94   9
head(SpatialExperiment::spatialCoords(spe_new))
#>                    pxl_col_in_fullres pxl_row_in_fullres
#> AAACAACGAATAGTTC-1               1874              50718
#> AAACAAGTATCTCCCA-1              13935              38805
#> AAACAATCTACTAGCA-1               2599              46977
#> AAACACCAATAACTGC-1              16101              50308
#> AAACAGAGCGACTCCT-1               5255              39910
#> AAACAGCTTTCAGAAG-1              12242              51692