#!/usr/bin/env Rscript --vanilla

##Check to see if necessary packages are installed
#CRAN packages
cran.packages <- c("optparse", "yaml", "igraph", "Rtsne", "spam", "MCMCprecision")

cran.package.check <- lapply(cran.packages, FUN = function(x) {
    if (!require(x, character.only = TRUE)) {
        install.packages(x, dependencies = TRUE)
    }
})

#Bioconductor packages
bioc.packages <- c("singleCellTK", "celda", "BiocParallel")

bioc.package.check <- lapply(bioc.packages, FUN = function(x) {
    if (!require(x, character.only = TRUE)) {
        BiocManager::install(x)
    }
})

## Function to parse arguments from yaml file
.parseConfig <- function(sctkConfig, arguments) {
  for (i in seq_along(arguments)) {
    arg <- arguments[i]
    assign(arg, sctkConfig[[arg]], envir = parent.frame())
  }
}

## Check whether python module is available
if (!reticulate::py_module_available(module = "scrublet")) {
    stop("Cannot find python module 'scrublet'. ",
            "Scrublet can be installed on the local machine",
            "with pip (e.g. pip install --user scanpy) and then the 'use_python()'",
            " function from the 'reticulate' package can be used to select the",
            " correct Python environment.")
}

##Read in flags from command line using optparse
option_list <- list(optparse::make_option(c("-b", "--basePath"),
        type="character",
        default=NULL,
        help="Base path for the output from the preprocessing algorithm"),
    optparse::make_option(c("-P", "--preproc"),
        type = "character",
        default=NULL,
        help="Algorithm used for preprocessing. One of 'CellRangerV2', 'CellRangerV3', 'BUStools', 'STARSolo', 'SEQC', 'Optimus', 'DropEst', 'SceRDS', 'CountMatrix', 'AnnData'"),
    optparse::make_option(c("-s","--sample"),
        type="character",
        help="Name of the sample. This will be prepended to the cell barcodes."),
    optparse::make_option(c("-o","--directory"),
        type="character",
        default=".",
        help="Output directory"),
    optparse::make_option(c("-O","--outputPrefix"),
        type="character",
        default="CombinedSamples",
        help="Prefix of the name of output file when --splitSample=FALSE."),
    optparse::make_option(c("-g","--gmt"),
        type="character",
        default=NULL,
        help="GMT file containing gene sets for quality control. The second column in the GMT file (i.e. the description) should contain the location to look for the IDs in the data. If set to 'rownames', then the gene set IDs will be matched with the row IDs of the data matrix. If another character or integer index is supplied, then gene set IDs will be matched to IDs the that column of feature table."),
    optparse::make_option(c("-t","--delim"),
        type="character",
        default="\t",
        help="Delimiter used in GMT file"),
    optparse::make_option(c("-G","--genome"),
        type="character",
        default=NULL,
        help="The name of genome reference. This is only required for CellRangerV2 data."), 
    optparse::make_option(c("-C","--cellPath"),
        type="character",
        default=NULL,
        help="The directory contains cell matrix, gene and cell barcodes information. Default is NULL. If 'basePath' is NULL, both 'cellPath' and 'rawPath' should also be specified."),
    optparse::make_option(c("-R","--rawPath"),
        type="character",
        default=NULL,
        help="The directory contains droplet matrix, gene and cell barcodes information. Default is NULL. If 'basePath' is NULL, both 'cellPath' and 'rawPath' should also be specified."),
    optparse::make_option(c("-S","--splitSample"),
        type="logical",
        default=TRUE,
        help="Save SingleCellExperiment object for each sample. Default is FALSE. If TRUE, all samples will be combined and only one combimed SingleCellExperiment object will be saved."),
    optparse::make_option(c("-r","--rawData"),
        type="character",
        default=NULL,
        help="The full path of the RDS file or Matrix file of the raw gene count matrix. This would be provided only when --preproc is SceRDS or CountMatrix."),
    optparse::make_option(c("-c","--cellData"),
        type="character",
        default=NULL,
        help="The full path of the RDS file or Matrix file of the cell count matrix. This would be use only when --preproc is SceRDS or CountMatrix."),
    optparse::make_option(c("-F", "--outputFormat"),
        type="character",
        default=NULL,
        help="The output format of this QC pipeline. Currently, it supports SCE, Flatfile, Seurat, AnnData and HTAN."),
    optparse::make_option(c("-y", "--yamlFile"),
        type="character",
        default=NULL,
        help="YAML file containing parameters called by singleCellTK QC functions. Please check documentation for details."),
    optparse::make_option(c("-d", "--dataType"),
        type="character",
        default="Both",
        help="Type of data as input. Default is Both, which means taking both droplet and cell matrix as input. If set as 'Droplet', it will only processes droplet data. If set as 'Cell', it will only processes cell data."),
    optparse::make_option(c("-n", "--numCores"),
        type="integer",
        default=1,
        help="Number of cores used to run the pipeline. By default is 1. Parallel computing is enabled if -n is greater than 1."),
    optparse::make_option(c("-D", "--detectCells"),
        type="logical",
        default=FALSE,
        help="Detect cells from droplet matrix. Default is FALSE. This argument is only eavluated when -d is 'Droplet'. If set as TRUE, cells will be detected and cell matrixed will be subset from the droplet matrix. Also, quality control will be performed on the detected cell matrix."),
    optparse::make_option(c("-m", "--cellDetectMethod"),
        type="character",
        default='EmptyDrops',
        help="Methods to detect cells. Default is 'EmptyDrops'. Other options could be 'Knee' or 'Inflection'. More information is provided in the documentation. "),
    optparse::make_option(c("-i", "--studyDesign"),
        type="character",
        default=NULL,
        help="The txt file containing the desrciption of the study design. Default is NULL. This would be shown at the begining the html report of cell and droplet QC."),
    optparse::make_option(c("-L", "--subTitle"),
        type="character",
        default=NULL,
        help="The subtitle used in the cell and droplet QC HTML report. Default is None. The subtitle can contain information of the sample, like sample name, etc. The length of subsitle should be the same as the length of samples, if -S is set as TRUE. if -S is set as FALSE, the length of subtitle should be one or NULL"),
    optparse::make_option(c("-T", "--parallelType"),
        type="character",
        default="MulticoreParam",
        help="Type of clusters used for parallel computing. Default is 'MulticoreParam'. It can be 'MulticoreParam' or 'SnowParam'. This argument will be evaluated only when numCores > 1.")
    )
## Define arguments
arguments <- optparse::parse_args(optparse::OptionParser(option_list=option_list), positional_arguments=TRUE)
opt <- arguments$options
process <- unlist(strsplit(opt[["preproc"]], ","))
sample <- unlist(strsplit(opt[["sample"]], ","))
directory <- unlist(strsplit(opt[["directory"]], ","))
gmt <- opt[["gmt"]]
sep <- opt[["delim"]]
split <- opt[["splitSample"]]
basepath <- opt[["basePath"]]
FilterDir <- opt[["cellPath"]] 
RawDir <- opt[["rawPath"]]
Reference <- opt[["genome"]]
RawFile <- opt[["rawData"]]
FilterFile <- opt[["cellData"]]
yamlFile <- opt[["yamlFile"]]
formats <- opt[["outputFormat"]]
dataType <- opt[["dataType"]]
detectCell <- opt[["detectCells"]]
numCores <- opt[["numCores"]]
parallelType <- opt[["parallelType"]] 
cellCalling <- opt[["cellDetectMethod"]]
studyDesign <- opt[["studyDesign"]]
subTitles <- opt[["subTitle"]]
CombinedSamplesName <- opt[["outputPrefix"]]

if (!is.null(basepath)) { basepath <- unlist(strsplit(opt[["basePath"]], ",")) } 

if (!is.null(FilterDir)) { FilterDir <- unlist(strsplit(opt[["cellPath"]], ",")) } 

if (!is.null(RawDir)) { RawDir <- unlist(strsplit(opt[["rawPath"]], ",")) } 

if (!is.null(Reference)) { Reference <- unlist(strsplit(opt[["genome"]], ",")) } 

if (!is.null(RawFile)) { RawFile <- unlist(strsplit(opt[["rawData"]], ",")) }

if (!is.null(FilterFile)) { FilterFile <- unlist(strsplit(opt[["cellData"]], ",")) } 

if (!is.null(formats)) { formats <- unlist(strsplit(opt[["outputFormat"]], ",")) } 

if (!is.null(studyDesign)) { studyDesign <- base::readLines(studyDesign, n=-1) }

if (is.null(subTitles)) { 
    subTitles <- paste("SCTK QC HTML report for sample", sample)
} else {
    subTitles <- unlist(strsplit(opt[["subTitles"]], ","))
}

## Parse parameters for QC algorithms
if (!is.null(yamlFile)) {
    arguments <- c('Params')
    qcParams <- yaml::read_yaml(yamlFile)
    .parseConfig(qcParams, arguments)
} else {
    Params <- list()
}

## checking numCores argument
isWindows <- .Platform$OS.type == "windows"

if (numCores > 1) {
    if (numCores > parallel::detectCores()) {
        warning("numCores is greater than number of cores available. Set numCores as maximum number of cores available.")
    }

    numCores <- min(numCores, parallel::detectCores())
    message(as.character(numCores), " cores are used for parallel computation.")

    if (parallelType == "MulticoreParam") {
        parallelParam <- MulticoreParam(workers = numCores)

        if (isTRUE(isWindows)) {
            warning("'MulticoreParam' is not supported for Windows system. Setting 'parallelType' as 'SnowParam'. ")
            parallelParam <- SnowParam(workers = numCores)            
        }

    } else if (parallelType == "SnowParam") {
        parallelParam <- SnowParam(workers = numCores)
    } else {
        stop("'--parallelType' should be 'MulticoreParam' or 'SnowParam'.")
    }

    Params$QCMetrics$BPPARAM <- parallelParam
    Params$emptyDrops$BPPARAM <- parallelParam
    #Params$doubletCells$BPPARAM <- parallelParam
    Params$doubletFinder$nCores <- numCores
}

### checking output formats
if (!all(formats %in% c("SCE", "AnnData", "FlatFile", "HTAN", "Seurat"))) {
    warning("Output format must be 'SCE', 'AnnData', 'HTAN', 'Seurat' or 'FlatFile'. Format ", 
         paste(formats[!formats %in% c("SCE", "AnnData", "FlatFile", "HTAN", "Seurat")], collapse = ","),
         " is not supported now. ")
}

formats <- formats[formats %in% c("SCE", "AnnData", "FlatFile", "HTAN", "Seurat")]
message("The output format is [", 
        paste(formats, collapse = ","), "]. ")

if (length(formats) == 0) {
    warning("None of the provided format is supported now. Therefore, the output ", 
        "will be SCE, AnnData, Seurat, FlatFile and HTAN. ")
    formats <- c("SCE", "AnnData", "FlatFile", "HTAN", "Seurat")
}

if (!(dataType %in% c("Both", "Droplet", "Cell"))) {
    stop("-d / -dataType must be one of the following: 'Both', 'Droplet' or 'Cell'. ")
}

## Checking argument
if (dataType == "Both") {
    if (is.null(RawFile) & is.null(FilterFile)) {
        if (is.null(basepath)) {
            if ((is.null(FilterDir) || is.null(RawDir))) {
                stop("Both 'cellPath' and 'rawPath' need to be specified when 'basePath' is NULL.")
            } else {
                # message("'basePath' is NULL. Data is loaded using directories specified by '--cell_data_path' and '--raw_data_path'.")
                if (length(FilterDir) != length(RawDir)) {
                    stop("The length of '--cellPath' should be the same as the length of '--rawPath'.")
                }
                if (length(FilterDir) != length(sample)) {
                    stop("The length of '--cellPath' should be the same as the length of '--sample'.")
                }
                if (length(FilterDir) != length(process)) {
                    stop('The length of "--cellPath" should be the same as ',
                             'the length of "--preproc"!')
                }
            }
        } else {
            if (length(basepath) != length(process)) {
                stop('The length of "--basePath" should be the same as ',
                         'the length of "--preproc"!')
            }
            if (length(basepath) != length(sample)) {
                stop('The length of "--basePath" should be the same as ',
                         'the length of "--sample"!')   

            if (length(Reference) != sum(process == 'CellRangerV2')) {
                stop('The length of --genome should be the same as ',
                        'the number of "CellRangerV2" in the "--preproc"!')        
        }  
            }
        }

       
    }

    if (!is.null(RawFile) | !is.null(FilterFile)) {
        if (length(RawFile) != length(FilterFile)) {
             stop("The length of '--rawData' and '--cellData' should be the same when '--preproc' is SceRDS or CountMatrix.")
        }
        if (length(FilterFile) != length(sample)) {
            stop("The length of '--cellData' should be the same as the length of '--sample'.")
        }
        if (length(FilterFile) != length(process)) {
            stop('The length of "--cellData" should be the same as ',
                     'the length of "--preproc"!')
        }
    }
}

if (dataType == "Cell") {
    if (is.null(FilterFile)) {
        if (is.null(basepath)) {
            if ((is.null(FilterDir))) {
                stop("'cellPath' need to be specified when 'basePath' is NULL.")
            } 
            # message("'base_path' is NULL. Data is loaded using directories specified by '--cell_data_path' and '--raw_data_path'.")
            if (length(FilterDir) != length(sample)) {
                stop("The length of '--cellPath' should be the same as the length of '--sample'.")
            }
            if (length(FilterDir) != length(process)) {
                stop('The length of "--cellPath" should be the same as ',
                         'the length of "--preproc"!')
            }
            
        } else {
            if (length(basepath) != length(process)) {
                stop('The length of "--basePath" should be the same as ',
                         'the length of "--preproc"!')
            }
            if (length(basepath) != length(sample)) {
                stop('The length of "--basePath" should be the same as ',
                         'the length of "--sample"!')    
            }
            if (length(Reference) != sum(process == 'CellRangerV2')) {
                stop('The length of --genome should be the same as ',
                        'the number of "CellRangerV2" in the "--preproc"!')        
            }
        }

    }

    if (!is.null(FilterFile)) {
        if (length(FilterFile) != length(sample)) {
            stop("The length of '--cellData' should be the same as the length of '--sample'.")
        }
        if (length(FilterFile) != length(process)) {
            stop('The length of "--cellData" should be the same as ',
                     'the length of "--preproc"!')
        }
    }
}

if (dataType == "Droplet") {
    if (is.null(RawFile)) {
        if (is.null(basepath)) {
            if ((is.null(RawDir))) {
                stop("'rawPath' need to be specified when 'basePath' is NULL.")
            } 
            # message("'base_path' is NULL. Data is loaded using directories specified by '--cell_data_path' and '--raw_data_path'.")
            if (length(RawDir) != length(sample)) {
                stop("The length of '--rawPath' should be the same as the length of '--sample'.")
            }
            if (length(RawDir) != length(process)) {
                stop('The length of "--rawPath" should be the same as ',
                         'the length of "--preproc"!')
            }
            
        } else {
            if (length(basepath) != length(process)) {
                stop('The length of "--basePath" should be the same as ',
                         'the length of "--preproc"!')
            }
            if (length(basepath) != length(sample)) {
                stop('The length of "--basePath" should be the same as ',
                         'the length of "--sample"!')    
            }
            if (length(Reference) != sum(process == 'CellRangerV2')) {
                stop('The length of --genome should be the same as ',
                        'the number of "CellRangerV2" in the "--preproc"!')        
            }
        }       
    }

    if (!is.null(RawFile)) {
        if (length(RawFile) != length(sample)) {
            stop("The length of '--rawData' should be the same as the length of '--sample'.")
        }
        if (length(RawFile) != length(process)) {
            stop('The length of "--rawData" should be the same as ',
                     'the length of "--preproc"!')
        }
    }
}

if (!cellCalling %in% c("Knee", "Inflection", "EmptyDrops")) {
    stop("The --cellDetectMethod must be 'Knee', 'Inflection' or 'Emptydrops'.")
}

## Prepare for QC
dropletSCE_list <- list()
cellSCE_list <- list()
geneSetCollection <- NULL
if (!is.null(gmt)) {
    geneSetCollection <- GSEABase::getGmt(gmt, sep=sep)
}


level3Meta <- list()
level4Meta <- list()

for(i in seq_along(process)) {
    preproc <- process[i]
    samplename <- sample[i]
    path <- basepath[i]
    raw <- RawDir[i]
    fil <- FilterDir[i]
    ref <- Reference[i]
    rawFile <- RawFile[i]
    filFile <- FilterFile[i]
    subTitle <- subTitles[i]
    INPUT <- qcInputProcess(preproc,
                            samplename,
                            path,
                            raw,
                            fil,
                            ref,
                            rawFile,
                            filFile,
                            dataType)

    dropletSCE <- INPUT[[1]]
    cellSCE <- INPUT[[2]]
    
    if (dataType == "Cell") {
        if (is.null(cellSCE) && (preproc %in% c("BUStools", "SEQC"))) {
            dropletSCE <- runDropletQC(inSCE = dropletSCE, paramsList=Params)
            ix <- !is.na(dropletSCE$dropletUtils_emptyDrops_fdr) & (dropletSCE$dropletUtils_emptyDrops_fdr < 0.01)
            cellSCE <- dropletSCE[,ix]
        }

        message(paste0(date(), " .. Running cell QC"))        
        cellSCE <- runCellQC(inSCE = cellSCE, geneSetCollection = geneSetCollection, paramsList=Params)
    }

    if (dataType == "Droplet") {
        message(paste0(date(), " .. Running droplet QC"))        
        dropletSCE <- runDropletQC(inSCE = dropletSCE, paramsList=Params)
        if (isTRUE(detectCell)) {
            if (cellCalling == "EmptyDrops") {
                ix <- !is.na(dropletSCE$dropletUtils_emptyDrops_fdr) & dropletSCE$dropletUtils_emptyDrops_fdr < 0.01 
            } else if (cellCalling == "Knee") {
                ix <- dropletSCE$dropletUtils_BarcodeRank_Knee == 1                 
            } else {
                ix <- dropletSCE$dropletUtils_BarcodeRank_Inflection == 1
            }
            cellSCE <- dropletSCE[,ix]
            message(paste0(date(), " .. Running cell QC"))
            cellSCE <- runCellQC(inSCE = cellSCE, geneSetCollection = geneSetCollection, paramsList=Params)
        }
    }

    if (dataType == "Both") {
        if (!is.null(dropletSCE)) {
            message(paste0(date(), " .. Running droplet QC"))        
            dropletSCE <- runDropletQC(inSCE = dropletSCE, paramsList=Params)
            
            if (is.null(cellSCE)) {
                if (cellCalling == "EmptyDrops") {
                    ix <- !is.na(dropletSCE$dropletUtils_emptyDrops_fdr) & dropletSCE$dropletUtils_emptyDrops_fdr < 0.01 
                } else if (cellCalling == "Knee") {
                    ix <- dropletSCE$dropletUtils_BarcodeRank_Knee == 1                 
                } else {
                    ix <- dropletSCE$dropletUtils_BarcodeRank_Inflection == 1
                }
                cellSCE <- dropletSCE[,ix]
            }    
        }

        if (!is.null(cellSCE)) {
            message(paste0(date(), " .. Running cell QC"))        
            cellSCE <- runCellQC(inSCE = cellSCE, geneSetCollection = geneSetCollection, paramsList=Params)
        }

        cbInCellMat <- colnames(dropletSCE) %in% colnames(cellSCE)
        SummarizedExperiment::colData(dropletSCE)$barcodeInCellMatrix <- cbInCellMat
    }
    
    ## merge colData of dropletSCE and FilteredSCE
    mergedDropletSCE <- NULL
    mergedFilteredSCE <- NULL

    if (dataType == "Both") {
        mergedDropletSCE <- mergeSCEColData(dropletSCE, cellSCE)
        mergedFilteredSCE <- mergeSCEColData(cellSCE, dropletSCE)
    }

    if (dataType == "Cell") {
        mergedFilteredSCE <- cellSCE
    }

    if (dataType == "Droplet") {
        if (isTRUE(detectCell)) {
            mergedDropletSCE <- mergeSCEColData(dropletSCE, cellSCE)
            mergedFilteredSCE <- mergeSCEColData(cellSCE, dropletSCE)            
        } else{
            mergedDropletSCE <- dropletSCE
        }
    }
    

    if (isTRUE(split)) {
        ### assign sample to every runBarcodeRanksMetaOutput metadata slot
        if (!is.null(mergedDropletSCE)) {
            names(metadata(mergedDropletSCE)$runBarcodeRanksMetaOutput) <- samplename   
        }

        if (!is.null(mergedFilteredSCE)) {
            for (name in names(metadata(mergedFilteredSCE))) {
                metadata(mergedFilteredSCE)[[name]] <- list(metadata(mergedFilteredSCE)[[name]])
                names(metadata(mergedFilteredSCE)[[name]]) <- samplename
            }
        }
        
        if ((dataType == "Both") | (dataType == "Droplet" & isTRUE(detectCell))) {
            exportSCE(inSCE = mergedDropletSCE, samplename = samplename, directory = directory, type = "Droplets", format=formats)
            exportSCE(inSCE = mergedFilteredSCE, samplename = samplename, directory = directory, type = "Cells", format=formats)
            

            ## Get parameters of QC functions
            getSceParams(inSCE = mergedFilteredSCE, directory = directory, 
                         samplename = samplename, writeYAML = TRUE,
                         skip = c("scrublet", "runDecontX", "runBarcodeRanksMetaOutput"))

            ## generate meta data
            if ("FlatFile" %in% formats) {
                if ("HTAN" %in% formats) {
                    meta <- generateMeta(dropletSCE = mergedDropletSCE, cellSCE = mergedFilteredSCE, samplename = samplename, 
                                        dir = directory, HTAN=TRUE, dataType = "Both")
                } else {
                    meta <- generateMeta(dropletSCE = mergedDropletSCE, cellSCE = mergedFilteredSCE, samplename = samplename, 
                                        dir = directory, HTAN=FALSE, dataType = "Both")  
                }

                level3Meta[[i]] <- meta[[1]]
                level4Meta[[i]] <- meta[[2]]

            } else {
                warning("'FlatFile' is not in output format. Skip exporting the manifest file.")
            }

            ## generate html report
            reportDropletQC(inSCE = mergedDropletSCE, output_dir = directory, output_file = paste0("SCTK_", samplename,'_dropletQC.html'), subTitle = subTitle, studyDesign = studyDesign)
            reportCellQC(inSCE = mergedFilteredSCE, output_dir = directory, output_file = paste0("SCTK_", samplename,'_cellQC.html'), subTitle = subTitle, studyDesign = studyDesign)

            ## generate QC metrics table for mergedFilteredSCE
            QCsummary <- sampleSummaryStats(mergedFilteredSCE, simple=FALSE, sample = colData(mergedFilteredSCE)$sample) #colData(cellSCE)$Study_ID
            write.csv(QCsummary, file.path(directory, 
                                           samplename,
                                           paste0("SCTK_", samplename,'_cellQC_summary.csv')))
        }

        if ((dataType == "Droplet") & (!isTRUE(detectCell))) {
            exportSCE(inSCE = mergedDropletSCE, samplename = samplename, directory = directory, type = "Droplets", format=formats)
            if ("FlatFile" %in% formats) {
                if ("HTAN" %in% formats) {
                    meta <- generateMeta(dropletSCE = mergedDropletSCE, cellSCE = NULL, samplename = samplename, 
                                        dir = directory, HTAN=TRUE, dataType = "Droplet")
                } else {
                    meta <- generateMeta(dropletSCE = mergedDropletSCE, cellSCE = NULL, samplename = samplename, 
                                        dir = directory, HTAN=FALSE, dataType = "Droplet")  
                }

                level3Meta[[i]] <- meta[[1]]
                level4Meta[[i]] <- meta[[2]]

            } else {
                warning("'FlatFile' is not in output format. Skip exporting the manifest file.")
            }

            reportDropletQC(inSCE = mergedDropletSCE, output_dir = directory, output_file = paste0("SCTK_", samplename,'_dropletQC.html'), subTitle = subTitle, studyDesign = studyDesign)
        }

        if (dataType == "Cell") {
            exportSCE(inSCE = mergedFilteredSCE, samplename = samplename, directory = directory, type = "Cells", format=formats)
            if ("FlatFile" %in% formats) {
                if ("HTAN" %in% formats) {
                    meta <- generateMeta(dropletSCE = NULL, cellSCE = mergedFilteredSCE, samplename = samplename, 
                                        dir = directory, HTAN=TRUE, dataType = "Cell")
                } else {
                    meta <- generateMeta(dropletSCE = NULL, cellSCE = mergedFilteredSCE, samplename = samplename, 
                                        dir = directory, HTAN=FALSE, dataType = "Cell")  
                }

                level3Meta[[i]] <- meta[[1]]
                level4Meta[[i]] <- meta[[2]]

            } else {
                warning("'FlatFile' is not in output format. Skip exporting the manifest file.")
            }

            reportCellQC(inSCE = mergedFilteredSCE, output_dir = directory, output_file = paste0("SCTK_", samplename,'_cellQC.html'), subTitle = subTitle, studyDesign = studyDesign)

            getSceParams(inSCE = mergedFilteredSCE, directory = directory, 
                         samplename = samplename, writeYAML = TRUE,
                         skip = c("scrublet", "runDecontX", "runBarcodeRanksMetaOutput"))

            QCsummary <- sampleSummaryStats(mergedFilteredSCE, simple=FALSE, sample = colData(mergedFilteredSCE)$sample) #colData(cellSCE)$Study_ID
            write.csv(QCsummary, file.path(directory, 
                                           samplename,
                                           paste0("SCTK_", samplename,'_cellQC_summary.csv')))
        }

    }
    dropletSCE_list[[samplename]] <- mergedDropletSCE
    cellSCE_list[[samplename]] <- mergedFilteredSCE
}

if (!isTRUE(split)) {
    if (length(sample) > 1) {
        samplename <- CombinedSamplesName #paste(sample, collapse="-")
        subTitle <- paste("SCTK QC HTML report for sample", samplename)
    }

    if ((dataType == "Both") | (dataType == "Droplet" & isTRUE(detectCell))) {
        by.r <- NULL
        by.c <- Reduce(intersect, lapply(dropletSCE_list, function(x) { colnames(colData(x))}))
        dropletSCE <- combineSCE(dropletSCE_list, by.r, by.c, combined = TRUE)
        names(metadata(dropletSCE)$runBarcodeRanksMetaOutput) <- sample

        by.c <- Reduce(intersect, lapply(cellSCE_list, function(x) { colnames(colData(x))}))
        cellSCE <- combineSCE(cellSCE_list, by.r, by.c, combined = TRUE)
        for (name in names(metadata(cellSCE))) {
            names(metadata(cellSCE)[[name]]) <- sample
        }

        exportSCE(inSCE = dropletSCE, samplename = samplename, directory = directory, type = "Droplets", format=formats)
        exportSCE(inSCE = cellSCE, samplename = samplename, directory = directory, type = "Cells", format=formats)

        ## html report
        reportDropletQC(inSCE = dropletSCE, output_dir = directory, output_file = paste0("SCTK_", samplename,'_dropletQC.html'), subTitle = subTitle, studyDesign = studyDesign)
        reportCellQC(inSCE = cellSCE, output_dir = directory, output_file = paste0("SCTK_", samplename,'_cellQC.html'), subTitle = subTitle, studyDesign = studyDesign)

        ## Get parameters of QC functions
        getSceParams(inSCE = cellSCE, directory = directory, samplename = samplename, writeYAML = TRUE)

        ## generate meta data
        if ("FlatFile" %in% formats) {
            if ("HTAN" %in% formats) {
                meta <- generateMeta(dropletSCE = dropletSCE, cellSCE = cellSCE, samplename = samplename, 
                                    dir = directory, HTAN=TRUE, dataType = "Both")
            } else {
                meta <- generateMeta(dropletSCE = dropletSCE, cellSCE = cellSCE, samplename = samplename, 
                                    dir = directory, HTAN=FALSE, dataType = "Both")            
            }

            level3Meta <- list(meta[[1]])
            level4Meta <- list(meta[[2]])

        } else {
            warning("'FlatFile' is not in output format. Skip exporting the manifest file.")
        }

        ## generate QC summary
        QCsummary <- sampleSummaryStats(cellSCE, simple=FALSE, sample = colData(cellSCE)$sample) 
        write.csv(QCsummary, file.path(directory,
                                       samplename, 
                                       paste0("SCTK_", samplename,'_cellQC_summary.csv')))
    }

    if (dataType == "Cell") {
        by.r <- NULL
        by.c <- Reduce(intersect, lapply(cellSCE_list, function(x) { colnames(colData(x))}))

        cellSCE <- combineSCE(cellSCE_list, by.r, by.c, combined = TRUE)
        for (name in names(metadata(cellSCE))) {
            names(metadata(cellSCE)[[name]]) <- sample
        }

        exportSCE(inSCE = cellSCE, samplename = samplename, directory = directory, type = "Cells", format=formats)
        if ("FlatFile" %in% formats) {
            if ("HTAN" %in% formats) {
                meta <- generateMeta(dropletSCE = NULL, cellSCE = cellSCE, samplename = samplename, 
                                    dir = directory, HTAN=TRUE, dataType = "Cell")
            } else {
                meta <- generateMeta(dropletSCE = NULL, cellSCE = cellSCE, samplename = samplename, 
                                    dir = directory, HTAN=FALSE, dataType = "Cell")  
            }

            level3Meta[[i]] <- meta[[1]]
            level4Meta[[i]] <- meta[[2]]

        } else {
            warning("'FlatFile' is not in output format. Skip exporting the manifest file.")
        }

        reportCellQC(inSCE = cellSCE, output_dir = directory, output_file = paste0("SCTK_", samplename,'_cellQC.html'), subTitle = subTitle, studyDesign = studyDesign)
        getSceParams(inSCE = cellSCE, directory = directory, samplename = samplename, writeYAML = TRUE)

        QCsummary <- sampleSummaryStats(cellSCE, simple=FALSE, sample = colData(cellSCE)$sample) 
        write.csv(QCsummary, file.path(directory,
                                       samplename, 
                                       paste0("SCTK_", samplename,'_cellQC_summary.csv')))
    }

    if ((dataType == "Droplet") & (!isTRUE(detectCell))) {
        by.r <- NULL
        by.c <- Reduce(intersect, lapply(dropletSCE_list, function(x) { colnames(colData(x))}))
        dropletSCE <- combineSCE(dropletSCE_list, by.r, by.c, combined = TRUE)
        names(metadata(dropletSCE)$runBarcodeRanksMetaOutput) <- sample
        
        exportSCE(inSCE = dropletSCE, samplename = samplename, directory = directory, type = "Droplets", format=formats)
        if ("FlatFile" %in% formats) {
            if ("HTAN" %in% formats) {
                meta <- generateMeta(dropletSCE = dropletSCE, cellSCE = NULL, samplename = samplename, 
                                    dir = directory, HTAN=TRUE, dataType = "Droplet")
            } else {
                meta <- generateMeta(dropletSCE = dropletSCE, cellSCE = NULL, samplename = samplename, 
                                    dir = directory, HTAN=FALSE, dataType = "Droplet")  
            }

            level3Meta[[i]] <- meta[[1]]
            level4Meta[[i]] <- meta[[2]]

        } else {
            warning("'FlatFile' is not in output format. Skip exporting the manifest file.")
        }

        reportDropletQC(inSCE = dropletSCE, output_dir = directory, output_file = paste0("SCTK_", samplename,'_dropletQC.html'), subTitle = subTitle, studyDesign = studyDesign)
    
    }
}

if (("FlatFile" %in% formats)) {
    HTANLevel3 <- do.call(base::rbind, level3Meta)
    HTANLevel4 <- do.call(base::rbind, level4Meta)
    write.csv(HTANLevel3, file = file.path(directory, "level3Meta.csv"))
    #if ((dataType == "Both") | (dataType == "Droplet" & isTRUE(detectCell))) {
    if ( !(dataType == "Droplet" & !isTRUE(detectCell)) ) {
        write.csv(HTANLevel4, file = file.path(directory, "level4Meta.csv"))
    }
}

sessionInfo()
