R/GetThresholdedIntensities.R

In RGLab/COMPASS: Combinatorial Polyfunctionality Analysis of Single Cells

##' Extract Thresholded Intensities from a GatingSet
##'
##' This function extracts thresholded intensities for children of a
##' node \code{node}, as specified through the \code{map} argument.
##'
##' \code{map} should be an \R \code{list}, mapping node names (as
##' specified in the gating hierarchy of the gating set) to channel
##' names (as specified in either the \code{desc} or \code{name}
##' columns of the parameters of the associated \code{flowFrame}s
##' in the \code{GatingSet}).
##'
##' @param gs A \code{GatingSet} or \code{GatingSetList}.
##' @param node The name, or path, of a single node in a
##'   \code{GatingSet} / \code{GatingSetList}.
##' @param map A \code{list}, mapping node names to markers.
##' @example examples/GetThresholdedIntensities.R
##' @return A \code{list} with two components:
##'
##' \item{data}{A \code{list} of thresholded intensity measures.}
##' \item{counts}{A named vector of total cell counts at the node \code{node}.}
##'
##' @export
GetThresholdedIntensities <- function(gs, node, map) {

  if (requireNamespace("flowWorkspace",quietly = TRUE)) {

    node_names <- names(map)
    channel_names <- unlist( unname(map) )

    if (!(inherits(gs, "GatingSet") || inherits(gs, "GatingSetList"))) {
      stop("'gs' must be either a 'GatingSet' or a 'GatingSetList'")
    }

    ## Turn the gs into a list of gs's
    if (inherits(gs, "GatingSet")) {
      gslist <- vector("list", length(gs))
      for (i in 1:length(gs)) {
        gslist[[i]] <- gs[[i]]
      }
      names(gslist) <- flowWorkspace::sampleNames(gs)
    } else {
      stop("Internal Error: not implemented for GatingSetLists yet.", call.=FALSE)
    }

    ## Make 'node' act more like a regular expression if it isn't one already
    ## But only do this if it's not '/' or 'root'
    if (!(node %in% c("root", "/"))) {
      n <- nchar(node)
      if (!substring(node, 1, 1) == "/") node <- paste0("/", node)
      if (!substring(node, n, n) == "$") node <- paste0(node, "$")
      node <- gsub("(?<!\\\\)\\+", "\\\\+", node, perl=TRUE)
    }

    paths <- flowWorkspace::gh_get_pop_paths(gslist[[1]])
    path <- paths[ grepl(node, paths, fixed = FALSE) ]

    if (length(path) > 1) {
      stop(gettextf("The node expression %s is not unique.", node))
    }

    if (length(path) == 0) {
      stop(gettextf("The node expression %s doesn't identify any nodes.",
        node))
    }

    # Extract the parent node name from the full population name
    node_name <- gsub(".*/", "", path)
    message(gettextf("Fetching data from children of '%s'.", path))

    # extract all the counts
    message("Extracting cell counts")
    counts <- unlist(lapply(gslist, function(x) {
      flowWorkspace::gh_pop_get_count(x, path)
    }))

    # Get the children of that parent and filter out boolean gates Test if
    # children exist, and test if non-empty set returned.
    message("Looking for expected children nodes")
    child.nodes <- flowWorkspace::gh_pop_get_children(gs[[1]], node_name)
    child.nodes <- basename(child.nodes)
    if (length(child.nodes) == 0) {
      stop(gettextf("Population %s has no children! Choose a different parent population.",
        node_name))
    }

    child.nodes <- child.nodes[ !sapply(child.nodes, function(x)
      gh_pop_is_bool_gate(gs[[1]], x))]

    if (length(child.nodes) == 0) {
      stop(gettextf("All the children of %s are boolean gates. Choose a population with non-boolean child gates.",
        node_name))
    }

    message("We will extract data from the following nodes:\n\n",
      paste( strwrap( paste(child.nodes, collapse=", "), 60), collapse="\n" ))

    ## This next block of code tries to figure out which channel names we will be using

    ## Try to guess whether we should be pulling names from the 'desc'
    ## column or the 'name' column of the flowSets
    ff <- flowWorkspace::gh_pop_get_data(gs[[1]], use.exprs=FALSE)
    params <- flowCore::parameters(ff)@data

    ## First, check for a perfect match using a basic regex
    .check_match <- function(x, vec) {
      all( sapply(channel_names, function(x) {
        any( x == na.omit(vec) )
      }) )
    }

    matched_flag <- FALSE ## did we match everything in 'map' to the channel names?
    column_to_use <- NULL ## are we going to use 'desc' or 'name'?
    indices <- NULL ## what indices are we using to pull from the flowFrame?

    if (!all(params$name == colnames( flowCore::exprs(ff) ))) {
      stop("Internal Error: expected 'params$desc' and 'colnames( exprs(ff) )' to ",
        "be identical but they are not!", call.=FALSE)
    }

    if (.check_match(channel_names, params$desc)) {
      message("Channel names were matched to the 'desc' column.")
      matched_flag <- TRUE
      column_to_use <- "desc"
      indices <- match(channel_names, params$desc)
    }

    if (!matched_flag && .check_match(channel_names, params$name)) {
      message("Channel names were matched to the 'name' column.")
      matched_flag <- TRUE
      column_to_use <- "name"
      indices <- match(channel_names, params$name)
    }

    if (!matched_flag) {
      stop("Unable to match values in 'names(map)' to channel names in the data.")
    }

    expr_nms <- unname(as.character(params[[column_to_use]])[indices])

    ## Extract the intensities
    message("Extracting cell intensities and thresholding...")
    intensities <- lapply(gslist, function(x) {
      exprs <- flowCore::exprs( flowWorkspace::gh_pop_get_data(x, path) )[, expr_nms, drop=FALSE]
      for (i in seq_along(node_names)) {
        cNode <- node_names[i]
        cChannel <- channel_names[i]
        ## Need to handle 'root' specially
        if (identical(path, "root")) {
          node_path <- paste0("/", cNode)
        } else {
          node_path <- file.path(path, cNode)
        }
        ## find out what cells didn't fall into the gate
        ind <- !flowWorkspace::gh_pop_get_indices(x, node_path)
        exprs[ind, expr_nms[i]] <- 0
      }
      exprs <- exprs[ rowSums(exprs) > 0, , drop=FALSE]
      return(exprs)
    })

    if (!all(names(counts) == names(intensities))) {
      stop("Internal Error: expected `!all(names(counts) == names(intensities))` to be TRUE.", call.=FALSE)
    }

    message("Done!")
    return( list(
      data=intensities,
      counts=counts
    ) )

  }

  .needsFlowWorkspace()

}