From 842af6b5bf2337d27403e1286accb791fe192661 Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Sat, 17 May 2025 01:37:00 +0000 Subject: [PATCH 01/24] variable uniformity, main workflow to subworkflows, fixed stub runs --- conf/test.config | 41 +++++- modules/local/baysor/run/main.nf | 2 + modules/local/spatialdata/merge/main.nf | 7 +- modules/local/spatialdata/meta/main.nf | 22 +-- modules/local/spatialdata/write/main.nf | 10 +- .../local/baysor_generate_preview/main.nf | 6 +- .../baysor_run_morphology_ome_tif/main.nf | 17 +-- .../local/baysor_run_transcripts_csv/main.nf | 11 +- .../main.nf | 14 +- .../main.nf | 22 +-- .../local/proseg_preset_proseg2baysor/main.nf | 8 +- .../local/segger_create_train_predict/main.nf | 2 +- .../spatialdata_write_meta_merge/main.nf | 14 +- .../main.nf | 12 +- .../xeniumranger_relabel_resegment/main.nf | 16 ++- .../main.nf | 11 +- workflows/spatialxe.nf | 135 ++++++++---------- 17 files changed, 201 insertions(+), 149 deletions(-) diff --git a/conf/test.config b/conf/test.config index 77b495da..9398ab12 100644 --- a/conf/test.config +++ b/conf/test.config @@ -14,9 +14,9 @@ process { withLabel: process_high { resourceLimits = [ - cpus: 8, - memory: '8.GB', - time: '1.h' + cpus: 8, + memory: '8.GB', + time: '1.h' ] } @@ -26,6 +26,41 @@ process { memory: '8.GB' ] } + + withName: BAYSOR_RUN { + resourceLimits = [ + cpus: 4, + memory: '8.GB' + ] + } + + withName: XENIUMRANGER_IMPORT_SEGMENTATION { + resourceLimits = [ + cpus: 4, + memory: '8.GB' + ] + } + + withName: PROSEG { + resourceLimits = [ + cpus: 4, + memory: '8.GB' + ] + } + + withName: PROSEG2BAYSOR { + resourceLimits = [ + cpus: 4, + memory: '8.GB' + ] + } + + withName: SPATIALDATA_WRITE { + resourceLimits = [ + cpus: 4, + memory: '8.GB' + ] + } } params { diff --git a/modules/local/baysor/run/main.nf b/modules/local/baysor/run/main.nf index 619880a2..455308e2 100644 --- a/modules/local/baysor/run/main.nf +++ b/modules/local/baysor/run/main.nf @@ -58,10 +58,12 @@ process BAYSOR_RUN { """ touch segmentation.csv touch segmentation_polygons_2d.json + touch segmentation_polygons_3d.json touch segmentation_log.log touch segmentation_counts.loom touch segmentation_cell_stats.csv touch segmentation_params.dump.toml + touch segmentation_run.html cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/spatialdata/merge/main.nf b/modules/local/spatialdata/merge/main.nf index 528e540d..dbcc00af 100644 --- a/modules/local/spatialdata/merge/main.nf +++ b/modules/local/spatialdata/merge/main.nf @@ -6,11 +6,11 @@ process SPATIALDATA_MERGE { input: tuple val(meta), path(ref_bundle, stageAs: "*") - tuple val(meta), path(add_bundle, stageAs: "*") + path(add_bundle, stageAs: "*") output: - tuple val(meta), path("spatialdata_spatialxe") , emit: spatialxe_bundle - path "versions.yml" , emit: versions + tuple val(meta), path("spatialdata_spatialxe"), emit: spatialxe_bundle + path("versions.yml") , emit: versions when: task.ext.when == null || task.ext.when @@ -22,6 +22,7 @@ process SPATIALDATA_MERGE { } def args = task.ext.args ?: '' + template 'merge.py' stub: diff --git a/modules/local/spatialdata/meta/main.nf b/modules/local/spatialdata/meta/main.nf index 4f302df4..260372aa 100644 --- a/modules/local/spatialdata/meta/main.nf +++ b/modules/local/spatialdata/meta/main.nf @@ -4,30 +4,32 @@ process SPATIALDATA_META { container "heylf/spatialdata:0.2.6" - // Exit if running this module with -profile conda / -profile mamba - if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) { - exit 1, "SPATIALDATA_WRITE module does not support Conda. Please use Docker / Singularity / Podman instead." - } - input: tuple val(meta), path(spatialdata_bundle, stageAs: "*") - tuple val(meta), path(xenium_bundle, stageAs: "*") + path(xenium_bundle, stageAs: "*") output: - tuple val(meta), path("spatialdata_spatialxe_final") , emit: spatialxe_bundle - path "versions.yml" , emit: versions + tuple val(meta), path("spatialdata_spatialxe_final"), emit: spatialxe_bundle + path("versions.yml") , emit: versions when: task.ext.when == null || task.ext.when script: + // Exit if running this module with -profile conda / -profile mamba + if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) { + exit 1, "SPATIALDATA_META module does not support Conda. Please use Docker / Singularity / Podman instead." + } + def args = task.ext.args ?: '' + template 'meta.py' stub: + """ - mkdir -p "spatialdata_spatialxe/" - touch spatialdata_spatialxe/fake_file.txt + mkdir -p "spatialdata_spatialxe_final/" + touch "spatialdata_spatialxe_final/fake_file.txt" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/modules/local/spatialdata/write/main.nf b/modules/local/spatialdata/write/main.nf index 19dd9abc..53d8cbb6 100644 --- a/modules/local/spatialdata/write/main.nf +++ b/modules/local/spatialdata/write/main.nf @@ -9,8 +9,8 @@ process SPATIALDATA_WRITE { val(outputfolder) output: - tuple val(meta), path("${outputfolder}") , emit: spatialdata - path "versions.yml" , emit: versions + tuple val(meta), path("${outputfolder}"), emit: spatialdata + path("versions.yml") , emit: versions when: task.ext.when == null || task.ext.when @@ -26,9 +26,11 @@ process SPATIALDATA_WRITE { template 'write.py' stub: + + def outdir = "${outputfolder}" """ - mkdir -p "spatialdata/" - touch spatialdata/fake_file.txt + mkdir -p "${outdir}/" + touch "${outdir}/fake_file.txt" cat <<-END_VERSIONS > versions.yml "${task.process}": diff --git a/subworkflows/local/baysor_generate_preview/main.nf b/subworkflows/local/baysor_generate_preview/main.nf index 62e41bff..59f3b810 100644 --- a/subworkflows/local/baysor_generate_preview/main.nf +++ b/subworkflows/local/baysor_generate_preview/main.nf @@ -10,8 +10,8 @@ workflow BAYSOR_GENERATE_PREVIEW { take: - ch_transcripts // channel: [ val(meta), ["transcripts.csv.gz"] ] - ch_config // channel: ["path-to-xenium.toml"] + ch_transcripts_csv // channel: [ val(meta), ["path-to-transcripts.csv.gz"] ] + ch_config // channel: ["path-to-xenium.toml"] main: @@ -20,7 +20,7 @@ workflow BAYSOR_GENERATE_PREVIEW { // unzip transcripts.csv.gz - GUNZIP ( ch_transcripts ) + GUNZIP ( ch_transcripts_csv ) ch_versions = ch_versions.mix ( GUNZIP.out.versions ) ch_unzipped_transcripts = GUNZIP.out.gunzip diff --git a/subworkflows/local/baysor_run_morphology_ome_tif/main.nf b/subworkflows/local/baysor_run_morphology_ome_tif/main.nf index 86a42f1e..94c78a9b 100644 --- a/subworkflows/local/baysor_run_morphology_ome_tif/main.nf +++ b/subworkflows/local/baysor_run_morphology_ome_tif/main.nf @@ -12,10 +12,10 @@ workflow BAYSOR_RUN_MORPHOLOGY_OME_TIF { take: - ch_bundle // channel: [ val(meta), ["xenium-bundle"] ] - ch_transcripts // channel: [ val(meta), ["transcripts.csv.gz"] ] - ch_image // channel: [ val(meta), ["morphology_focus.tiff"] ] - ch_config // channel: ["path-to-xenium.toml"] + ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] + ch_transcripts_csv // channel: [ val(meta), ["path-to-transcripts.csv.gz"] ] + ch_morphology_image // channel: [ val(meta), ["path-to-morphology_focus.tiff"] ] + ch_config // channel: ["path-to-xenium.toml"] main: @@ -31,7 +31,7 @@ workflow BAYSOR_RUN_MORPHOLOGY_OME_TIF { // unzip transcripts.csv.gz - GUNZIP ( ch_transcripts ) + GUNZIP ( ch_transcripts_csv ) ch_versions = ch_versions.mix ( GUNZIP.out.versions ) ch_unzipped_transcripts = GUNZIP.out.gunzip @@ -40,7 +40,7 @@ workflow BAYSOR_RUN_MORPHOLOGY_OME_TIF { ch_just_image = Channel.empty() if ( params.sharpen_tiff ) { - RESOLIFT ( ch_image ) + RESOLIFT ( ch_morphology_image ) ch_versions = ch_versions.mix( RESOLIFT.out.versions ) ch_enhanced_tiff = RESOLIFT.out.enhanced_tiff @@ -51,7 +51,7 @@ workflow BAYSOR_RUN_MORPHOLOGY_OME_TIF { } else { // use the original morphology tiff from the bundle - ch_just_image = ch_image.map { + ch_just_image = ch_morphology_image.map { _meta, image -> return [ image ] } } @@ -71,9 +71,10 @@ workflow BAYSOR_RUN_MORPHOLOGY_OME_TIF { } ch_polygons2d = BAYSOR_RUN_IMAGE.out.polygons2d ch_htmls = BAYSOR_RUN_IMAGE.out.htmls + // run xeniumranger import-segmentation XENIUMRANGER_IMPORT_SEGMENTATION ( - ch_bundle, + ch_bundle_path, [], [], [], diff --git a/subworkflows/local/baysor_run_transcripts_csv/main.nf b/subworkflows/local/baysor_run_transcripts_csv/main.nf index 386b55f7..10b067d8 100644 --- a/subworkflows/local/baysor_run_transcripts_csv/main.nf +++ b/subworkflows/local/baysor_run_transcripts_csv/main.nf @@ -11,10 +11,9 @@ workflow BAYSOR_RUN_TRANSCRIPTS_CSV { take: - ch_bundle // channel: [ val(meta), ["xenium-bundle"] ] - ch_transcripts // channel: [ val(meta), ["transcripts.csv.gz"] ] - ch_image // channel: [ val(meta), ["morphology_focus.tiff"] ] - ch_config // channel: ["path-to-xenium.toml"] + ch_bundle_path // channel: [ val(meta), ["xenium-bundle"] ] + ch_transcripts_csv // channel: [ val(meta), ["transcripts.csv.gz"] ] + ch_config // channel: ["path-to-xenium.toml"] main: @@ -29,7 +28,7 @@ workflow BAYSOR_RUN_TRANSCRIPTS_CSV { // unzip transcripts.csv.gz - GUNZIP ( ch_transcripts ) + GUNZIP ( ch_transcripts_csv ) ch_versions = ch_versions.mix ( GUNZIP.out.versions ) ch_unzipped_transcripts = GUNZIP.out.gunzip @@ -52,7 +51,7 @@ workflow BAYSOR_RUN_TRANSCRIPTS_CSV { // run xeniumranger import-segmentation XENIUMRANGER_IMPORT_SEGMENTATION ( - ch_bundle, + ch_bundle_path, [], [], [], diff --git a/subworkflows/local/cellpose_baysor_import_segmentation/main.nf b/subworkflows/local/cellpose_baysor_import_segmentation/main.nf index 7abf8454..22f556b9 100644 --- a/subworkflows/local/cellpose_baysor_import_segmentation/main.nf +++ b/subworkflows/local/cellpose_baysor_import_segmentation/main.nf @@ -10,17 +10,17 @@ workflow CELLPOSE_BAYSOR_IMPORT_SEGMENTATION { take: - ch_image // channel: [ val(meta), ["path-to-morphology.ome.tif"] ] - ch_bundle // channel: [ val(meta), ["path-to-xenium-bundle"] ] - ch_transcripts // channel: [ val(meta), ["path-to-transcripts.parquet"] ] - ch_config // channel: ["path-to-xenium.toml"] + ch_morphology_image // channel: [ val(meta), ["path-to-morphology.ome.tif"] ] + ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] + ch_transcripts_parquet // channel: [ val(meta), ["path-to-transcripts.parquet"] ] + ch_config // channel: ["path-to-xenium.toml"] main: ch_versions = Channel.empty() // run cellpose to generate segmentation mask - CELLPOSE ( ch_image, []) + CELLPOSE ( ch_morphology_image, []) ch_versions = ch_versions.mix ( CELLPOSE.out.versions ) @@ -28,7 +28,7 @@ workflow CELLPOSE_BAYSOR_IMPORT_SEGMENTATION { ch_mask = CELLPOSE.out.mask.map { _meta, seg_mask -> [ seg_mask ] } - BAYSOR_RUN ( ch_transcripts, ch_mask, ch_config, 30 ) + BAYSOR_RUN ( ch_transcripts_parquet, ch_mask, ch_config, 30 ) ch_versions = ch_versions.mix ( BAYSOR_RUN.out.versions ) @@ -39,7 +39,7 @@ workflow CELLPOSE_BAYSOR_IMPORT_SEGMENTATION { ch_polygons = BAYSOR_RUN.out.polygons2d XENIUMRANGER_IMPORT_SEGMENTATION ( - ch_bundle, + ch_bundle_path, [], [], [], diff --git a/subworkflows/local/cellpose_resolift_morphology_ome_tif/main.nf b/subworkflows/local/cellpose_resolift_morphology_ome_tif/main.nf index 1b20888f..46007d5e 100644 --- a/subworkflows/local/cellpose_resolift_morphology_ome_tif/main.nf +++ b/subworkflows/local/cellpose_resolift_morphology_ome_tif/main.nf @@ -10,27 +10,32 @@ workflow CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF { take: - ch_image // channel: [ val(meta), ["morphology.ome.tiff"] ] - ch_bundle // channel: [ val(meta), ["xenium-bundle"] ] + ch_morphology_image // channel: [ val(meta), ["path-to-morphology.ome.tiff"] ] + ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] main: ch_versions = Channel.empty() + ch_model = Channel.empty() + + if ( !params.cellpose_model ) { + ch_model = "" + } // sharpen morphology tiff if param - sharpen_tiff is true if ( params.sharpen_tiff ) { - RESOLIFT ( ch_image ) + RESOLIFT ( ch_morphology_image ) ch_versions = ch_versions.mix( RESOLIFT.out.versions ) // run cellpose on the enhanced tiff - CELLPOSE ( RESOLIFT.out.enhanced_tiff, params.cellpose_model ) + CELLPOSE ( RESOLIFT.out.enhanced_tiff, ch_model ) ch_versions = ch_versions.mix( CELLPOSE.out.versions ) } else { // run cellpose on the original tiff - CELLPOSE ( ch_image, params.cellpose_model ) + CELLPOSE ( ch_morphology_image, params.cellpose_model ) ch_versions = ch_versions.mix( CELLPOSE.out.versions ) } @@ -41,7 +46,7 @@ workflow CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF { cellpose_mask = CELLPOSE.out.mask.map { _meta, mask -> return [ mask ] } - cellpose_flows = CELLPOSE.out.flows.map { + _cellpose_flows = CELLPOSE.out.flows.map { _meta, flows -> return [ flows ] } @@ -49,7 +54,7 @@ workflow CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF { if ( params.nucleus_segmentation_only ) { XENIUMRANGER_IMPORT_SEGMENTATION ( - ch_bundle, + ch_bundle_path, [], cellpose_mask, [], @@ -58,10 +63,11 @@ workflow CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF { "" ) ch_versions = ch_versions.mix( XENIUMRANGER_IMPORT_SEGMENTATION.out.versions ) + } else { XENIUMRANGER_IMPORT_SEGMENTATION ( - ch_bundle, + ch_bundle_path, [], cellpose_mask, cellpose_cells, diff --git a/subworkflows/local/proseg_preset_proseg2baysor/main.nf b/subworkflows/local/proseg_preset_proseg2baysor/main.nf index 6c3a2441..20c90f64 100644 --- a/subworkflows/local/proseg_preset_proseg2baysor/main.nf +++ b/subworkflows/local/proseg_preset_proseg2baysor/main.nf @@ -10,15 +10,15 @@ workflow PROSEG_PRESET_PROSEG2BAYSOR { take: - ch_bundle // channel: [ val(meta), ["xenium-bundle"] ] - ch_transcripts // channel: [ val(meta), [ "transcripts.csv.gz" ] ] + ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] + ch_transcripts_csv // channel: [ val(meta), [ "transcripts.csv.gz" ] ] main: ch_versions = Channel.empty() // run proseg with the xenium format - PROSEG ( ch_transcripts ) + PROSEG ( ch_transcripts_csv ) ch_versions = ch_versions.mix( PROSEG.out.versions ) @@ -33,7 +33,7 @@ workflow PROSEG_PRESET_PROSEG2BAYSOR { // run xeniumranger import-segmentation XENIUMRANGER_IMPORT_SEGMENTATION ( - ch_bundle, + ch_bundle_path, [], [], [], diff --git a/subworkflows/local/segger_create_train_predict/main.nf b/subworkflows/local/segger_create_train_predict/main.nf index 77b232e4..3c1f14bd 100644 --- a/subworkflows/local/segger_create_train_predict/main.nf +++ b/subworkflows/local/segger_create_train_predict/main.nf @@ -11,7 +11,7 @@ workflow SEGGER_CREATE_TRAIN_PREDICT { take: - ch_basedir // channel: [ val(meta), [ basedir ] ] + ch_basedir // channel: [ val(meta), [ "basedir" ] ] ch_transcripts_parquet // channel: [ val(meta), [bundle + "/transcripts.parquet"]] main: diff --git a/subworkflows/local/spatialdata_write_meta_merge/main.nf b/subworkflows/local/spatialdata_write_meta_merge/main.nf index 18b20289..1cff0812 100644 --- a/subworkflows/local/spatialdata_write_meta_merge/main.nf +++ b/subworkflows/local/spatialdata_write_meta_merge/main.nf @@ -10,7 +10,7 @@ include { SPATIALDATA_META } from '../../ workflow SPATIALDATA_WRITE_META_MERGE { take: - ch_raw_bundle // channel: [ val(meta), [ "xenium-bundle" ] ] + ch_bundle_path // channel: [ val(meta), [ "path-to-xenium-bundle" ] ] ch_redefined_bundle // channel: [ val(meta), [ "redefined-xenium-bundle" ] ] main: @@ -18,7 +18,7 @@ workflow SPATIALDATA_WRITE_META_MERGE { ch_versions = Channel.empty() // write spatialdata object from the raw xenium bundle - raw_bundle_path = ch_raw_bundle.map { meta, file_path -> + raw_bundle_path = ch_bundle_path.map { meta, file_path -> return [ meta, file(file_path) ] } SPATIALDATA_WRITE_RAW_BUNDLE ( @@ -40,17 +40,23 @@ workflow SPATIALDATA_WRITE_META_MERGE { // merge raw & redefined spatialdata objects + ch_just_redefined_bundle = ch_redefined_bundle.map { + _meta, bundle -> return [ bundle ] + } SPATIALDATA_MERGE_RAW_REDEFINED ( SPATIALDATA_WRITE_RAW_BUNDLE.out.spatialdata, - SPATIALDATA_WRITE_REDEFINED_BUNDLE.out.spatialdata + ch_just_redefined_bundle ) ch_versions = ch_versions.mix ( SPATIALDATA_MERGE_RAW_REDEFINED.out.versions ) // write metadata with spatialdata object + ch_just_bundle_path = ch_bundle_path.map { + _meta, bundle -> return [ bundle ] + } SPATIALDATA_META ( SPATIALDATA_MERGE_RAW_REDEFINED.out.spatialxe_bundle, - ch_raw_bundle + ch_just_bundle_path ) ch_versions = ch_versions.mix ( SPATIALDATA_META.out.versions ) diff --git a/subworkflows/local/xeniumranger_import_segmentation_redefine_bundle/main.nf b/subworkflows/local/xeniumranger_import_segmentation_redefine_bundle/main.nf index f1054b2e..461751ef 100644 --- a/subworkflows/local/xeniumranger_import_segmentation_redefine_bundle/main.nf +++ b/subworkflows/local/xeniumranger_import_segmentation_redefine_bundle/main.nf @@ -11,14 +11,14 @@ workflow XENIUMRANGER_IMPORT_SEGMENTATION_REDEFINE_BUNDLE { take: - ch_bundle // channel: [ val(meta), [ "xenium-bundle" ] ] + ch_bundle_path // channel: [ val(meta), [ "path-to-xenium-bundle" ] ] main: ch_versions = Channel.empty() ch_redefined_bundle = Channel.empty() - cells = ch_bundle.map { + cells = ch_bundle_path.map { _meta, bundle -> return [ bundle + "/cells.zarr.zip" ] } @@ -26,7 +26,7 @@ workflow XENIUMRANGER_IMPORT_SEGMENTATION_REDEFINE_BUNDLE { if ( params.expansion_distance == 0 || params.expansion_distance != 5 ){ IMP_SEG_COUNT_MATRIX_EXP_DISTANCE ( - ch_bundle, + ch_bundle_path, [], cells, [], @@ -43,7 +43,7 @@ workflow XENIUMRANGER_IMPORT_SEGMENTATION_REDEFINE_BUNDLE { if ( params.qupath_polygons && params.nucleus_segmentation_only ) { IMP_SEG_POLYGON_GEOJSON_INPUT ( - ch_bundle, + ch_bundle_path, [], params.qupath_polygons, [], @@ -58,7 +58,7 @@ workflow XENIUMRANGER_IMPORT_SEGMENTATION_REDEFINE_BUNDLE { } else if ( params.qupath_polygons ) { IMP_SEG_POLYGON_GEOJSON_INPUT ( - ch_bundle, + ch_bundle_path, [], params.qupath_polygons, params.qupath_polygons, @@ -80,7 +80,7 @@ workflow XENIUMRANGER_IMPORT_SEGMENTATION_REDEFINE_BUNDLE { if ( params.qupath_polygons && params.alignment_csv ) { IMP_SEG_TRANS_MATRIX_INPUT ( - ch_bundle, + ch_bundle_path, params.alignment_csv, params.qupath_polygons, params.qupath_polygons, diff --git a/subworkflows/local/xeniumranger_relabel_resegment/main.nf b/subworkflows/local/xeniumranger_relabel_resegment/main.nf index 657db843..99075ccb 100644 --- a/subworkflows/local/xeniumranger_relabel_resegment/main.nf +++ b/subworkflows/local/xeniumranger_relabel_resegment/main.nf @@ -9,17 +9,23 @@ workflow XENIUMRANGER_RELABEL_RESEGMENT { take: - ch_bundle // channel: [ val(meta), [ xenium-bundle-path ] ] - ch_gene_panel // channel: [ ["gene_panel.json"] ] + ch_bundle_path // channel: [ val(meta), [ "path-to-xenium-bundle" ] ] + ch_gene_panel // channel: [ ["gene_panel.json"] ] main: ch_versions = Channel.empty() - XENIUMRANGER_RELABEL ( ch_bundle, ch_gene_panel ) + ch_bundle_path.view() + XENIUMRANGER_RELABEL ( + ch_bundle_path, + ch_gene_panel + ) ch_versions = ch_versions.mix ( XENIUMRANGER_RELABEL.out.versions ) - XENIUMRANGER_RESEGMENT ( XENIUMRANGER_RELABEL.out.bundle ) + XENIUMRANGER_RESEGMENT ( + XENIUMRANGER_RELABEL.out.bundle + ) ch_versions = ch_versions.mix ( XENIUMRANGER_RESEGMENT.out.versions ) @@ -27,5 +33,5 @@ workflow XENIUMRANGER_RELABEL_RESEGMENT { redefined_bundle = XENIUMRANGER_RESEGMENT.out.bundle // channel: [ val(meta), ["redefined-xenium-bundle"] ] - versions = ch_versions // channel: [ versions.yml ] + versions = ch_versions // channel: [ versions.yml ] } diff --git a/subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main.nf b/subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main.nf index 1000c165..fcdabd7a 100644 --- a/subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main.nf +++ b/subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main.nf @@ -9,26 +9,29 @@ workflow XENIUMRANGER_RESEGMENT_MORPHOLOGY_OME_TIF { take: - ch_bundle // channel: [ val(meta), ["xenium-bundle"] ] + ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] main: ch_versions = Channel.empty() // run resegment with changed config values - XENIUMRANGER_RESEGMENT ( ch_bundle ) + XENIUMRANGER_RESEGMENT ( ch_bundle_path ) ch_versions = ch_versions.mix( XENIUMRANGER_RESEGMENT.out.versions ) // run import segmentation to redine - cells = ch_bundle.map { + nuclei = XENIUMRANGER_RESEGMENT.out.bundle.map { + _meta, bundle -> return [ bundle + "/cells.zarr.zip" ] + } + cells = XENIUMRANGER_RESEGMENT.out.bundle.map { _meta, bundle -> return [ bundle + "/cells.zarr.zip" ] } XENIUMRANGER_IMPORT_SEGMENTATION ( XENIUMRANGER_RESEGMENT.out.bundle, [], - cells, + nuclei, cells, [], [], diff --git a/workflows/spatialxe.nf b/workflows/spatialxe.nf index b31db6ea..c9e2bc53 100644 --- a/workflows/spatialxe.nf +++ b/workflows/spatialxe.nf @@ -17,7 +17,7 @@ include { paramsSummaryMap } from 'plugin/nf-sch include { UNTAR } from '../modules/nf-core/untar/main' // testdata stagign subworkflow -include { STAGE_TESTDATA } from '../subworkflows/local/utils_stage_testdata/main' +// include { STAGE_TESTDATA } from '../subworkflows/local/utils_stage_testdata/main' // coordinate-based segmentation subworklfows include { SEGGER_CREATE_TRAIN_PREDICT } from '../subworkflows/local/segger_create_train_predict/main' @@ -70,7 +70,9 @@ workflow SPATIALXE { ch_bundle_path = Channel.empty() ch_raw_bundle = Channel.empty() ch_gene_panel = Channel.empty() + ch_transcripts_csv = Channel.empty() ch_transcripts_parquet = Channel.empty() + ch_morphology_image = Channel.empty() ch_redefined_bundle = Channel.empty() ch_config = Channel.empty() @@ -81,66 +83,54 @@ workflow SPATIALXE { ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ - // check if its a test run - if ( workflow.profile.contains('test') ) { - - STAGE_TESTDATA ( - ch_samplesheet - ) - - ch_raw_bundle = STAGE_TESTDATA.out.ch_raw_bundle - ch_transcripts = STAGE_TESTDATA.out.ch_transcripts_csv - ch_transcripts_parquet = STAGE_TESTDATA.out.ch_transcripts_parquet - ch_image = STAGE_TESTDATA.out.ch_image - ch_config = STAGE_TESTDATA.out.ch_config - - } else { - - // get samplesheet fields - ch_bundle_path = ch_samplesheet.map { meta, bundle, _image -> - return [ meta, file(bundle)] - } - - // get xenium bundle files - ch_bundle = ch_samplesheet.map { meta, bundle, _image -> - def bundle_files = file(bundle).toList().collect() - return [meta, bundle_files] - } + // get sample, xenium bundle and image path + ch_bundle_path = ch_samplesheet.map { meta, bundle, _image -> + return [ meta, bundle ] + } - // get transcript.csv.gz - ch_transcripts = ch_samplesheet.map { meta, bundle, _image -> - def transcripts_csv = file(bundle.replaceFirst(/\/$/, '') + "/transcripts.csv.gz") - return [ meta, transcripts_csv ] - } + // get all files in the xenium bundle in a channel + ch_bundle_files = ch_samplesheet.map + { meta, bundle_path, _image -> + def files = file("${bundle_path}/*").collect() + [meta, [files]] + } + + // get transcript.csv.gz from the xenium bundle + ch_transcripts_csv = ch_samplesheet.map { meta, bundle, _image -> + def transcripts_csv = file(bundle.replaceFirst(/\/$/, '') + "/transcripts.csv.gz") + return [ meta, transcripts_csv ] + } + // ch_transcripts_csv.view() - // get transcript.parquet - ch_transcripts_parquet = ch_samplesheet.map { meta, bundle, _image -> - def transcripts_parquet = file(bundle.replaceFirst(/\/$/, '') + "/transcripts.parquet") - return [ meta, transcripts_parquet ] - } + // get transcript.parquet from the xenium bundle + ch_transcripts_parquet = ch_samplesheet.map { meta, bundle, _image -> + def transcripts_parquet = file(bundle.replaceFirst(/\/$/, '') + "/transcripts.parquet") + return [ meta, transcripts_parquet ] + } + // ch_transcripts_parquet.view() - // get morphology.ome.tif - ch_image = ch_samplesheet.map { meta, bundle, image -> - def morphology_img = image ? file(image) : file(bundle.replaceFirst(/\/$/, '') + "/morphology.ome.tif") - return [ meta, morphology_img ] - } + // get morphology.ome.tif from the xenium bundle + ch_morphology_image = ch_samplesheet.map { meta, bundle, image -> + def morphology_img = image ? file(image) : file(bundle.replaceFirst(/\/$/, '') + "/morphology.ome.tif") + return [ meta, morphology_img ] + } + // ch_morphology_image.view() - // get baysor xenium config - ch_config = Channel.fromPath("${projectDir}/assets/config/xenium.toml", checkIfExists: true) + // get baysor xenium config + ch_config = Channel.fromPath("${projectDir}/assets/config/xenium.toml", checkIfExists: true) - // get gene_panel.json if provided with --gene_panel, sets relabel_genes to true - if (( params.gene_panel )) { + // get gene_panel.json if provided with --gene_panel, sets relabel_genes to true + if (( params.gene_panel )) { - params.relabel_genes = true - ch_gene_panel = Channel.fromPath(params.gene_panel, checkIfExists: true) + params.relabel_genes = true + ch_gene_panel = Channel.fromPath(params.gene_panel, checkIfExists: true) - } else { + } else { - // gene panel to use if only --relabel_genes is provided - ch_gene_panel = ch_samplesheet.map { meta, bundle, _image -> - def gene_panel = file(bundle.replaceFirst(/\/$/, '') + "/gene_panel.json") - return [ meta, gene_panel ] - } + // gene panel to use if only --relabel_genes is provided + ch_gene_panel = ch_samplesheet.map { meta, bundle, _image -> + def gene_panel = file(bundle.replaceFirst(/\/$/, '') + "/gene_panel.json") + return [ meta, gene_panel ] } } @@ -154,7 +144,7 @@ workflow SPATIALXE { if ( params.relabel_genes ) { XENIUMRANGER_RELABEL_RESEGMENT ( - ch_bundle, + ch_bundle_path, ch_gene_panel ) ch_raw_bundle = XENIUMRANGER_RELABEL_RESEGMENT.out.redefined_bundle @@ -172,7 +162,7 @@ workflow SPATIALXE { if ( params.generate_preview && params.mode == 'coordinate' ) { BAYSOR_GENERATE_PREVIEW ( - ch_transcripts, + ch_transcripts_csv, ch_config ) log.info "Preview generated at ${params.outdir}" @@ -190,7 +180,7 @@ workflow SPATIALXE { if ( !params.segmentation ) { CELLPOSE_BAYSOR_IMPORT_SEGMENTATION ( - ch_image, + ch_morphology_image, ch_bundle_path, ch_transcripts_parquet, ch_config @@ -205,7 +195,7 @@ workflow SPATIALXE { if ( params.segmentation == 'xeniumranger' ) { XENIUMRANGER_RESEGMENT_MORPHOLOGY_OME_TIF ( - ch_raw_bundle + ch_bundle_path ) ch_redefined_bundle = XENIUMRANGER_RESEGMENT_MORPHOLOGY_OME_TIF.out.redefined_bundle } @@ -214,9 +204,9 @@ workflow SPATIALXE { if ( params.segmentation == 'baysor' ) { BAYSOR_RUN_MORPHOLOGY_OME_TIF ( - ch_raw_bundle, - ch_transcripts, - ch_image, + ch_bundle_path, + ch_transcripts_csv, + ch_morphology_image, ch_config ) ch_redefined_bundle = BAYSOR_RUN_MORPHOLOGY_OME_TIF.out.redefined_bundle @@ -226,8 +216,8 @@ workflow SPATIALXE { if ( params.segmentation == 'cellpose' ) { CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF ( - ch_image, - ch_raw_bundle + ch_morphology_image, + ch_bundle_path ) ch_redefined_bundle = CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF.out.redefined_bundle } @@ -246,8 +236,8 @@ workflow SPATIALXE { if ( !params.segmentation ) { PROSEG_PRESET_PROSEG2BAYSOR ( - ch_raw_bundle, - ch_transcripts + ch_bundle_path, + ch_transcripts_csv ) ch_redefined_bundle = PROSEG_PRESET_PROSEG2BAYSOR.out.redefined_bundle @@ -260,8 +250,8 @@ workflow SPATIALXE { if ( params.segmentation == 'proseg') { PROSEG_PRESET_PROSEG2BAYSOR ( - ch_raw_bundle, - ch_transcripts + ch_bundle_path, + ch_transcripts_csv ) ch_redefined_bundle = PROSEG_PRESET_PROSEG2BAYSOR.out.redefined_bundle @@ -271,7 +261,7 @@ workflow SPATIALXE { if ( params.segmentation == 'segger' ) { SEGGER_CREATE_TRAIN_PREDICT ( - ch_raw_bundle, + ch_bundle_path, ch_transcripts_parquet ) @@ -281,10 +271,9 @@ workflow SPATIALXE { if ( params.segmentation == 'baysor' ) { BAYSOR_RUN_TRANSCRIPTS_CSV ( - ch_raw_bundle, - ch_transcripts, - ch_config, - [] + ch_bundle_path, + ch_transcripts_csv, + ch_config ) ch_redefined_bundle = BAYSOR_RUN_TRANSCRIPTS_CSV.out.redefined_bundle } @@ -301,7 +290,7 @@ workflow SPATIALXE { if ( params.xeniumranger_only ) { XENIUMRANGER_IMPORT_SEGMENTATION_REDEFINE_BUNDLE ( - ch_raw_bundle + ch_bundle_path ) ch_redefined_bundle = XENIUMRANGER_IMPORT_SEGMENTATION_REDEFINE_BUNDLE.out.redefined_bundle } @@ -314,7 +303,7 @@ workflow SPATIALXE { */ // run spatialdata modules to generate sd objects SPATIALDATA_WRITE_META_MERGE ( - ch_raw_bundle, + ch_bundle_path, ch_redefined_bundle ) From 0b4b945738266e617bba53b1886bde67aea538e6 Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Sat, 17 May 2025 16:08:24 +0000 Subject: [PATCH 02/24] manual testing changes --- nextflow.config | 4 +--- nextflow_schema.json | 11 +++++------ .../cellpose_resolift_morphology_ome_tif/main.nf | 9 +++------ 3 files changed, 9 insertions(+), 15 deletions(-) diff --git a/nextflow.config b/nextflow.config index 02db8443..d34b7f86 100644 --- a/nextflow.config +++ b/nextflow.config @@ -17,6 +17,7 @@ params { gene_panel = null // path to gene panel json file if `relabel_genes` is true qupath_polygons = null // polygon segmentation results in GeoJSON format alignment_csv = null // image alignment file format a 3x3 transformation matrix, where the last row is [0,0,1] + cellpose_model = null // custom cellpose model to use for running or starting training // execution specific sharpen_tiff = false // wether to sharpen the morphology-focus tiff @@ -36,9 +37,6 @@ params { segger_accelerator = 'cpu' // either 'cuda' or 'cpu' segger_knn_method = 'kd_tree' // 'cuda' - ensure your system has CUDA installed and configured properly - // Cellpose specific - cellpose_model = 'nuclei' // model to use for running or starting training default - cyto3 - // Proseg specific format = 'xenium' // preset value set as `xenium` diff --git a/nextflow_schema.json b/nextflow_schema.json index c1bd1276..78a77913 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -53,6 +53,11 @@ "description": "Image alignment file containing similarity transform matrix. (e.g., the _imagealignment.csv file exported from Xenium Explorer)", "format": "file-path" }, + "cellpose_model": { + "type": "string", + "description": "Model to use for running or starting training.", + "format": "file-path" + }, "email": { "type": "string", "description": "Email address for completion summary.", @@ -129,12 +134,6 @@ "enum": ["kd_tree", "cuda"], "description": "Method for KNN computation. (e.g., cuda for GPU-based computation)" }, - "cellpose_model": { - "type": "string", - "default": "nuclei", - "enum": ["nuclei", "cyto3"], - "description": "Model to use for running or starting training. (eg. cyto3 or nuclei)" - }, "format": { "type": "string", "default": "xenium", diff --git a/subworkflows/local/cellpose_resolift_morphology_ome_tif/main.nf b/subworkflows/local/cellpose_resolift_morphology_ome_tif/main.nf index 46007d5e..dd889264 100644 --- a/subworkflows/local/cellpose_resolift_morphology_ome_tif/main.nf +++ b/subworkflows/local/cellpose_resolift_morphology_ome_tif/main.nf @@ -16,11 +16,8 @@ workflow CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF { main: ch_versions = Channel.empty() - ch_model = Channel.empty() - if ( !params.cellpose_model ) { - ch_model = "" - } + cellpose_model = params.cellpose_model ? (Channel.fromPath(params.cellpose_model, checkIfExists: true)) : [] // sharpen morphology tiff if param - sharpen_tiff is true if ( params.sharpen_tiff ) { @@ -29,13 +26,13 @@ workflow CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF { ch_versions = ch_versions.mix( RESOLIFT.out.versions ) // run cellpose on the enhanced tiff - CELLPOSE ( RESOLIFT.out.enhanced_tiff, ch_model ) + CELLPOSE ( RESOLIFT.out.enhanced_tiff, cellpose_model ) ch_versions = ch_versions.mix( CELLPOSE.out.versions ) } else { // run cellpose on the original tiff - CELLPOSE ( ch_morphology_image, params.cellpose_model ) + CELLPOSE ( ch_morphology_image, cellpose_model ) ch_versions = ch_versions.mix( CELLPOSE.out.versions ) } From 49ce4ee5271db1f3f09c7c92d274a2b9dabc60ce Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Sat, 17 May 2025 16:21:33 +0000 Subject: [PATCH 03/24] manual testing changes --- conf/test.config | 14 ++++++++++++++ 1 file changed, 14 insertions(+) diff --git a/conf/test.config b/conf/test.config index 9398ab12..94c0e9f0 100644 --- a/conf/test.config +++ b/conf/test.config @@ -61,6 +61,20 @@ process { memory: '8.GB' ] } + + withName: SPATIALDATA_META { + resourceLimits = [ + cpus: 4, + memory: '8.GB' + ] + } + + withName: SPATIALDATA_MERGE { + resourceLimits = [ + cpus: 4, + memory: '8.GB' + ] + } } params { From 811fe5bf9ca0f49ddc7a3f97229af90f1490d5f8 Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Sun, 18 May 2025 00:34:46 +0000 Subject: [PATCH 04/24] manual testing changes --- conf/modules.config | 7 +++ nextflow.config | 1 + nextflow_schema.json | 5 +++ .../main.nf | 42 ++++-------------- .../spatialdata_write_meta_merge/main.nf | 14 ++---- .../utils_nfcore_spatialxe_pipeline/main.nf | 18 ++++++-- .../main.nf | 44 +++++++++++-------- workflows/spatialxe.nf | 16 ++++--- 8 files changed, 76 insertions(+), 71 deletions(-) rename subworkflows/local/{baysor_run_morphology_ome_tif => baysor_run_prior_segmentation_mask}/main.nf (64%) diff --git a/conf/modules.config b/conf/modules.config index 9c1c0373..254a9d1a 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -27,6 +27,13 @@ process { ] } + withName: XENIUMRANGER_RESEGMENT { + publishDir = [ + path: "${params.outdir}/xeniumranger/resegment", + mode: params.publish_dir_mode + ] + } + withName: XENIUMRANGER_IMPORT_SEGMENTATION { publishDir = [ path: "${params.outdir}/xeniumranger/import_segementation", diff --git a/nextflow.config b/nextflow.config index d34b7f86..68e9f593 100644 --- a/nextflow.config +++ b/nextflow.config @@ -18,6 +18,7 @@ params { qupath_polygons = null // polygon segmentation results in GeoJSON format alignment_csv = null // image alignment file format a 3x3 transformation matrix, where the last row is [0,0,1] cellpose_model = null // custom cellpose model to use for running or starting training + segmentation_mask = null // prior segmentation mask // execution specific sharpen_tiff = false // wether to sharpen the morphology-focus tiff diff --git a/nextflow_schema.json b/nextflow_schema.json index 78a77913..3b4749ce 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -58,6 +58,11 @@ "description": "Model to use for running or starting training.", "format": "file-path" }, + "segmentation_mask": { + "type": "string", + "description": "Prior segmentation mask from other segmentation methods.", + "format": "file-path" + }, "email": { "type": "string", "description": "Email address for completion summary.", diff --git a/subworkflows/local/baysor_run_morphology_ome_tif/main.nf b/subworkflows/local/baysor_run_prior_segmentation_mask/main.nf similarity index 64% rename from subworkflows/local/baysor_run_morphology_ome_tif/main.nf rename to subworkflows/local/baysor_run_prior_segmentation_mask/main.nf index 94c78a9b..a241f21c 100644 --- a/subworkflows/local/baysor_run_morphology_ome_tif/main.nf +++ b/subworkflows/local/baysor_run_prior_segmentation_mask/main.nf @@ -8,20 +8,19 @@ include { BAYSOR_RUN as BAYSOR_RUN_IMAGE } from '../../../modules/local/ba include { XENIUMRANGER_IMPORT_SEGMENTATION } from '../../../modules/nf-core/xeniumranger/import-segmentation/main' -workflow BAYSOR_RUN_MORPHOLOGY_OME_TIF { +workflow BAYSOR_RUN_PRIOR_SEGMENTATION_MASK { take: - ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] - ch_transcripts_csv // channel: [ val(meta), ["path-to-transcripts.csv.gz"] ] - ch_morphology_image // channel: [ val(meta), ["path-to-morphology_focus.tiff"] ] - ch_config // channel: ["path-to-xenium.toml"] + ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] + ch_transcripts_csv // channel: [ val(meta), ["path-to-transcripts.csv.gz"] ] + ch_segmentation_mask // channel: [ ["path-to-prior-segmentation-mask"] ] + ch_config // channel: ["path-to-xenium.toml"] main: ch_versions = Channel.empty() - ch_enhanced_tiff = Channel.empty() ch_segmentation = Channel.empty() ch_polygons2d = Channel.empty() ch_htmls = Channel.empty() @@ -29,44 +28,23 @@ workflow BAYSOR_RUN_MORPHOLOGY_OME_TIF { ch_redefined_bundle = Channel.empty() ch_unzipped_transcripts = Channel.empty() - // unzip transcripts.csv.gz GUNZIP ( ch_transcripts_csv ) ch_versions = ch_versions.mix ( GUNZIP.out.versions ) ch_unzipped_transcripts = GUNZIP.out.gunzip - // sharpen morphology tiff if param `sharpen_tiff` is true - ch_just_image = Channel.empty() - if ( params.sharpen_tiff ) { - - RESOLIFT ( ch_morphology_image ) - ch_versions = ch_versions.mix( RESOLIFT.out.versions ) - - ch_enhanced_tiff = RESOLIFT.out.enhanced_tiff - ch_just_image = ch_enhanced_tiff.map { - _meta, image -> return [ image ] - } - - } else { - - // use the original morphology tiff from the bundle - ch_just_image = ch_morphology_image.map { - _meta, image -> return [ image ] - } - } - // run baysor with morphology.tiff BAYSOR_RUN_IMAGE ( ch_unzipped_transcripts, - ch_just_image, + ch_segmentation_mask, ch_config, 30 ) ch_versions = ch_versions.mix( BAYSOR_RUN_IMAGE.out.versions ) ch_segmentation = BAYSOR_RUN_IMAGE.out.segmentation - ch_jus_segmentation = ch_segmentation.map { + ch_just_segmentation = ch_segmentation.map { _meta, segmentation -> return [ segmentation ] } ch_polygons2d = BAYSOR_RUN_IMAGE.out.polygons2d @@ -78,9 +56,9 @@ workflow BAYSOR_RUN_MORPHOLOGY_OME_TIF { [], [], [], - ch_jus_segmentation, + ch_just_segmentation, ch_polygons2d, - "pixel" + "microns" ) ch_versions = ch_versions.mix( XENIUMRANGER_IMPORT_SEGMENTATION.out.versions ) @@ -88,8 +66,6 @@ workflow BAYSOR_RUN_MORPHOLOGY_OME_TIF { emit: - enhanced_tiff = ch_enhanced_tiff // channel: [ val(meta), ["morphology.tiff"] ] - segmentation = ch_segmentation // channel: [ val(meta), ["segmentation.csv"] ] polygons2d = ch_polygons2d // channel: [ ["segmentation_polygons_2d.json"] ] htmls = ch_htmls // channel: [ ["*.html"] ] diff --git a/subworkflows/local/spatialdata_write_meta_merge/main.nf b/subworkflows/local/spatialdata_write_meta_merge/main.nf index 1cff0812..b032c785 100644 --- a/subworkflows/local/spatialdata_write_meta_merge/main.nf +++ b/subworkflows/local/spatialdata_write_meta_merge/main.nf @@ -18,29 +18,23 @@ workflow SPATIALDATA_WRITE_META_MERGE { ch_versions = Channel.empty() // write spatialdata object from the raw xenium bundle - raw_bundle_path = ch_bundle_path.map { meta, file_path -> - return [ meta, file(file_path) ] - } SPATIALDATA_WRITE_RAW_BUNDLE ( - raw_bundle_path, + ch_bundle_path, 'spatialdata_raw' ) ch_versions = ch_versions.mix ( SPATIALDATA_WRITE_RAW_BUNDLE.out.versions ) // write spatialdata object after running IMP_SEG - redefined_bundle_path = ch_redefined_bundle.map { meta, file_path -> - return [ meta, file(file_path) ] - } SPATIALDATA_WRITE_REDEFINED_BUNDLE ( - redefined_bundle_path, + ch_redefined_bundle, 'spatialdata_redefined' ) ch_versions = ch_versions.mix ( SPATIALDATA_WRITE_REDEFINED_BUNDLE.out.versions ) // merge raw & redefined spatialdata objects - ch_just_redefined_bundle = ch_redefined_bundle.map { + ch_just_redefined_bundle = SPATIALDATA_WRITE_REDEFINED_BUNDLE.out.spatialdata.map { _meta, bundle -> return [ bundle ] } SPATIALDATA_MERGE_RAW_REDEFINED ( @@ -67,6 +61,6 @@ workflow SPATIALDATA_WRITE_META_MERGE { ch_sd_merged = SPATIALDATA_MERGE_RAW_REDEFINED.out.spatialxe_bundle // channel: [ val(meta), "spatialdata_spatialxe" ] ch_sd_meta = SPATIALDATA_META.out.spatialxe_bundle // channel: [ val(meta), "spatialdata_spatialxe_final" ] - versions = ch_versions // channel: [ versions.yml ] + versions = ch_versions // channel: [ versions.yml ] } diff --git a/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf b/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf index 09c7ac0d..dceebf3d 100644 --- a/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf @@ -152,18 +152,30 @@ def validateInputParameters() { if ( params.mode == 'coordinate' && params.segmentation ) { if ( !params.transcript_seg_methods.contains(params.segmentation) ) { - error "Error: Invalid segmentation method: ${params.segmentation} provided for the `coordinate` based mode. Options: ${params.transcript_seg_methods}" + error "Error: Invalid segmentation method: `${params.segmentation}` provided for the `coordinate` based mode. Options: ${params.transcript_seg_methods}" } } // check if --relabel_genes is true but --gene_panel is not provided if ( params.relabel_genes && !params.gene_panel ) { - log.warn "Relabel genes is enabled, but gene panel is not provided with the `--gene_panel`. Using `gene_panel.json` in the xenium bundle" + log.warn "Relabel genes is enabled, but gene panel is not provided with the `--gene_panel`. Using `gene_panel.json` in the xenium bundle." } // check if --relabel_genes is true but --gene_panel is not provided if ( params.gene_panel && !params.relabel_genes ) { - log.warn "Gene panel provided, but relabel genes is disabled. Using `gene_panel.json` only to generate metadata" + log.warn "Gene panel provided, but relabel genes is disabled. Using `gene_panel.json` only to generate metadata." + } + + // check if segmentation method is xeniumranger and nucleus_ony_segmentation is enabled + if ( params.segmentation == 'xeniumranger' && !params.nucleus_segmentation_only ) { + log.warn "Nucleus segmentation is disabled. Running xeniumranger resegment module to redefine xenium bundle without nucleus segmentation." + log.warn "Use --nucleus_segmentation_only to enable nucleus segmentation to redefine xenium bundle with import-segmentation module." + } + + if ( params.mode == 'image' && params.segmentation == 'baysor' ) { + if ( !params.segmentation_mask ) { + error "Error: Missing path to segmentation mask. Image-based segmentation with the `baysor` method requires a segmentation mask with the `--segmentation_mask` option." + } } } diff --git a/subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main.nf b/subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main.nf index fcdabd7a..6907ff50 100644 --- a/subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main.nf +++ b/subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main.nf @@ -13,36 +13,44 @@ workflow XENIUMRANGER_RESEGMENT_MORPHOLOGY_OME_TIF { main: - ch_versions = Channel.empty() + ch_versions = Channel.empty() + ch_redefined_bundle = Channel.empty() // run resegment with changed config values XENIUMRANGER_RESEGMENT ( ch_bundle_path ) ch_versions = ch_versions.mix( XENIUMRANGER_RESEGMENT.out.versions ) - // run import segmentation to redine - nuclei = XENIUMRANGER_RESEGMENT.out.bundle.map { - _meta, bundle -> return [ bundle + "/cells.zarr.zip" ] - } - cells = XENIUMRANGER_RESEGMENT.out.bundle.map { + // run import segmentation to redine xenium bundle along with nuclear segmentation + cells = XENIUMRANGER_RESEGMENT.out.bundle.map { _meta, bundle -> return [ bundle + "/cells.zarr.zip" ] } - XENIUMRANGER_IMPORT_SEGMENTATION ( - XENIUMRANGER_RESEGMENT.out.bundle, - [], - nuclei, - cells, - [], - [], - "pixel" - ) - ch_versions = ch_versions.mix( XENIUMRANGER_IMPORT_SEGMENTATION.out.versions ) + // adjust the nuclear expansion distance without altering nuclei detection + if ( params.nucleus_segmentation_only ) { + + XENIUMRANGER_IMPORT_SEGMENTATION ( + XENIUMRANGER_RESEGMENT.out.bundle, + [], + cells, + [], + [], + [], + "pixels" + ) + ch_versions = ch_versions.mix( XENIUMRANGER_IMPORT_SEGMENTATION.out.versions ) + + ch_redefined_bundle = XENIUMRANGER_IMPORT_SEGMENTATION.out.bundle + + } else { + + ch_redefined_bundle = XENIUMRANGER_RESEGMENT.out.bundle + } emit: - redefined_bundle = XENIUMRANGER_IMPORT_SEGMENTATION.out.bundle // channel: [ val(meta), ["redefined-xenium-bundle"] ] + redefined_bundle = ch_redefined_bundle // channel: [ val(meta), ["redefined-xenium-bundle"] ] - versions = ch_versions // channel: [ versions.yml ] + versions = ch_versions // channel: [ versions.yml ] } diff --git a/workflows/spatialxe.nf b/workflows/spatialxe.nf index c9e2bc53..65106def 100644 --- a/workflows/spatialxe.nf +++ b/workflows/spatialxe.nf @@ -26,7 +26,7 @@ include { BAYSOR_GENERATE_PREVIEW } from '../subworkflo include { BAYSOR_RUN_TRANSCRIPTS_CSV } from '../subworkflows/local/baysor_run_transcripts_csv/main' // image-based segmentation subworklfows -include { BAYSOR_RUN_MORPHOLOGY_OME_TIF } from '../subworkflows/local/baysor_run_morphology_ome_tif/main' +include { BAYSOR_RUN_PRIOR_SEGMENTATION_MASK } from '../subworkflows/local/baysor_run_prior_segmentation_mask/main' include { CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF } from '../subworkflows/local/cellpose_resolift_morphology_ome_tif/main' include { CELLPOSE_BAYSOR_IMPORT_SEGMENTATION } from '../subworkflows/local/cellpose_baysor_import_segmentation/main' include { XENIUMRANGER_RESEGMENT_MORPHOLOGY_OME_TIF } from '../subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main' @@ -100,25 +100,27 @@ workflow SPATIALXE { def transcripts_csv = file(bundle.replaceFirst(/\/$/, '') + "/transcripts.csv.gz") return [ meta, transcripts_csv ] } - // ch_transcripts_csv.view() // get transcript.parquet from the xenium bundle ch_transcripts_parquet = ch_samplesheet.map { meta, bundle, _image -> def transcripts_parquet = file(bundle.replaceFirst(/\/$/, '') + "/transcripts.parquet") return [ meta, transcripts_parquet ] } - // ch_transcripts_parquet.view() // get morphology.ome.tif from the xenium bundle ch_morphology_image = ch_samplesheet.map { meta, bundle, image -> def morphology_img = image ? file(image) : file(bundle.replaceFirst(/\/$/, '') + "/morphology.ome.tif") return [ meta, morphology_img ] } - // ch_morphology_image.view() // get baysor xenium config ch_config = Channel.fromPath("${projectDir}/assets/config/xenium.toml", checkIfExists: true) + // get segmentation mask if provided with --segmentation_mask for the baysor method + if ( params.segmentation_mask ) { + ch_segmentation_mask = Channel.fromPath(params.segmentation_mask, checkIfExists: true) + } + // get gene_panel.json if provided with --gene_panel, sets relabel_genes to true if (( params.gene_panel )) { @@ -203,13 +205,13 @@ workflow SPATIALXE { // run baysor run with morphology_ome.tif if ( params.segmentation == 'baysor' ) { - BAYSOR_RUN_MORPHOLOGY_OME_TIF ( + BAYSOR_RUN_PRIOR_SEGMENTATION_MASK ( ch_bundle_path, ch_transcripts_csv, - ch_morphology_image, + ch_segmentation_mask, ch_config ) - ch_redefined_bundle = BAYSOR_RUN_MORPHOLOGY_OME_TIF.out.redefined_bundle + ch_redefined_bundle = BAYSOR_RUN_PRIOR_SEGMENTATION_MASK.out.redefined_bundle } // run cellpose on the morphology_ome.tif From 6792bced032f3d22101c2501b49ee4311ca8eb8c Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Sun, 18 May 2025 22:03:13 +0000 Subject: [PATCH 05/24] test profile fixes --- conf/test.config | 68 +--------- .../utils_nfcore_spatialxe_pipeline/main.nf | 13 ++ .../local/utils_stage_testdata/main.nf | 66 ---------- workflows/spatialxe.nf | 121 ++++++++++++------ 4 files changed, 98 insertions(+), 170 deletions(-) delete mode 100644 subworkflows/local/utils_stage_testdata/main.nf diff --git a/conf/test.config b/conf/test.config index 94c0e9f0..8e39243c 100644 --- a/conf/test.config +++ b/conf/test.config @@ -12,69 +12,11 @@ process { - withLabel: process_high { - resourceLimits = [ - cpus: 8, - memory: '8.GB', - time: '1.h' - ] - } - - withName: CELLPOSE { - resourceLimits = [ - cpus: 4, - memory: '8.GB' - ] - } - - withName: BAYSOR_RUN { - resourceLimits = [ - cpus: 4, - memory: '8.GB' - ] - } - - withName: XENIUMRANGER_IMPORT_SEGMENTATION { - resourceLimits = [ - cpus: 4, - memory: '8.GB' - ] - } - - withName: PROSEG { - resourceLimits = [ - cpus: 4, - memory: '8.GB' - ] - } - - withName: PROSEG2BAYSOR { - resourceLimits = [ - cpus: 4, - memory: '8.GB' - ] - } - - withName: SPATIALDATA_WRITE { - resourceLimits = [ - cpus: 4, - memory: '8.GB' - ] - } - - withName: SPATIALDATA_META { - resourceLimits = [ - cpus: 4, - memory: '8.GB' - ] - } - - withName: SPATIALDATA_MERGE { - resourceLimits = [ - cpus: 4, - memory: '8.GB' - ] - } + resourceLimits = [ + cpus: 8, + memory: '16.GB', + time: '1.h' + ] } params { diff --git a/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf b/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf index dceebf3d..bd13cea6 100644 --- a/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf @@ -78,7 +78,14 @@ workflow PIPELINE_INITIALISATION { } .set { ch_samplesheet } + + // + // Check and validate xenium bundle + // + validateXeniumBundle() + emit: + samplesheet = ch_samplesheet versions = ch_versions @@ -180,6 +187,12 @@ def validateInputParameters() { } +// +// Check and validate xenium bundle +// +def validateXeniumBundle() { + +} // // Generate methods description for MultiQC diff --git a/subworkflows/local/utils_stage_testdata/main.nf b/subworkflows/local/utils_stage_testdata/main.nf deleted file mode 100644 index 8a13590b..00000000 --- a/subworkflows/local/utils_stage_testdata/main.nf +++ /dev/null @@ -1,66 +0,0 @@ -// -// stage data for the test profile of the pipeline -// - -include { UNTAR } from '../../../modules/nf-core/untar/main' - - -workflow STAGE_TESTDATA { - - take: - ch_samplesheet - - main: - - ch_versions = Channel.empty() - ch_raw_bundle = Channel.empty() - ch_bundle_url = Channel.empty() - ch_image = Channel.empty() - ch_transcripts_csv = Channel.empty() - ch_transcripts_parquet = Channel.empty() - - // get xenium bundle path - ch_bundle_url = ch_samplesheet.map { meta, bundle, _image -> - return [ meta, file(bundle) ] - } - - // run the UNTAR module to create xenium bundle - UNTAR(ch_bundle_url) - ch_versions = ch_versions.mix ( UNTAR.out.versions ) - - ch_bundle_local_path = UNTAR.out.untar - ch_bundle_local_path.view() - - // get transcript.csv.gz - ch_transcripts_csv = ch_bundle_local_path.map { meta, bundle -> - def transcripts_csv = file(bundle + "/transcripts.csv.gz") - return [ meta, transcripts_csv ] - } - - // get transcript.parquet - ch_transcripts_parquet = ch_bundle_local_path.map { meta, bundle -> - def transcripts_parquet = file(bundle + "/transcripts.parquet") - return [ meta, transcripts_parquet ] - } - - // get morphology.ome.tif - ch_image = ch_bundle_local_path.map { meta, bundle -> - def morphology_image = file(bundle + "/morphology.ome.tif") - return [ meta, morphology_image ] - } - - // get baysor xenium config - ch_config = Channel.fromPath("${projectDir}/assets/config/xenium.toml", checkIfExists: true) - - - emit: - - ch_raw_bundle = ch_bundle_local_path // channel [ val(meta), ["xenium-bundle"] ] - ch_transcripts_csv = ch_transcripts_csv // channel [ val(meta), ["path-to-transcripts.csv.gz"] ] - ch_transcripts_parquet = ch_transcripts_parquet // channel [ val(meta), ["path-to-transcripts.csv.gz"] ] - ch_image = ch_image // channel [ val(meta), ["path-to-morphology.ome.tif"] ] - ch_config = ch_config // channel [ ["path-to-xenium.toml"] ] - - versions = ch_versions // channel [versions.yml] - -} diff --git a/workflows/spatialxe.nf b/workflows/spatialxe.nf index 65106def..5a1e7a50 100644 --- a/workflows/spatialxe.nf +++ b/workflows/spatialxe.nf @@ -5,42 +5,39 @@ */ // multiqc -include { MULTIQC } from '../modules/nf-core/multiqc/main' -include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' +include { MULTIQC } from '../modules/nf-core/multiqc/main' +include { paramsSummaryMultiqc } from '../subworkflows/nf-core/utils_nfcore_pipeline' // nf-core functionality -include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' -include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_spatialxe_pipeline' -include { paramsSummaryMap } from 'plugin/nf-schema' +include { softwareVersionsToYAML } from '../subworkflows/nf-core/utils_nfcore_pipeline' +include { methodsDescriptionText } from '../subworkflows/local/utils_nfcore_spatialxe_pipeline' +include { paramsSummaryMap } from 'plugin/nf-schema' // nf-core modules -include { UNTAR } from '../modules/nf-core/untar/main' - -// testdata stagign subworkflow -// include { STAGE_TESTDATA } from '../subworkflows/local/utils_stage_testdata/main' +include { UNTAR } from '../modules/nf-core/untar/main' // coordinate-based segmentation subworklfows -include { SEGGER_CREATE_TRAIN_PREDICT } from '../subworkflows/local/segger_create_train_predict/main' -include { PROSEG_PRESET_PROSEG2BAYSOR } from '../subworkflows/local/proseg_preset_proseg2baysor/main' -include { BAYSOR_GENERATE_PREVIEW } from '../subworkflows/local/baysor_generate_preview/main' -include { BAYSOR_RUN_TRANSCRIPTS_CSV } from '../subworkflows/local/baysor_run_transcripts_csv/main' +include { SEGGER_CREATE_TRAIN_PREDICT } from '../subworkflows/local/segger_create_train_predict/main' +include { PROSEG_PRESET_PROSEG2BAYSOR } from '../subworkflows/local/proseg_preset_proseg2baysor/main' +include { BAYSOR_GENERATE_PREVIEW } from '../subworkflows/local/baysor_generate_preview/main' +include { BAYSOR_RUN_TRANSCRIPTS_CSV } from '../subworkflows/local/baysor_run_transcripts_csv/main' // image-based segmentation subworklfows -include { BAYSOR_RUN_PRIOR_SEGMENTATION_MASK } from '../subworkflows/local/baysor_run_prior_segmentation_mask/main' -include { CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF } from '../subworkflows/local/cellpose_resolift_morphology_ome_tif/main' -include { CELLPOSE_BAYSOR_IMPORT_SEGMENTATION } from '../subworkflows/local/cellpose_baysor_import_segmentation/main' -include { XENIUMRANGER_RESEGMENT_MORPHOLOGY_OME_TIF } from '../subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main' +include { BAYSOR_RUN_PRIOR_SEGMENTATION_MASK } from '../subworkflows/local/baysor_run_prior_segmentation_mask/main' +include { CELLPOSE_RESOLIFT_MORPHOLOGY_OME_TIF } from '../subworkflows/local/cellpose_resolift_morphology_ome_tif/main' +include { CELLPOSE_BAYSOR_IMPORT_SEGMENTATION } from '../subworkflows/local/cellpose_baysor_import_segmentation/main' +include { XENIUMRANGER_RESEGMENT_MORPHOLOGY_OME_TIF } from '../subworkflows/local/xeniumranger_resegment_morphology_ome_tif/main' // segmentation-free subworkflows -include { BAYSOR_GENERATE_SEGFREE } from '../subworkflows/local/baysor_generate_segfree/main' -include { FICTURE_PREPROCESS_MODEL } from '../subworkflows/local/ficture_preprocess_model/main' +include { BAYSOR_GENERATE_SEGFREE } from '../subworkflows/local/baysor_generate_segfree/main' +include { FICTURE_PREPROCESS_MODEL } from '../subworkflows/local/ficture_preprocess_model/main' // xeniumranger subworkflows include { XENIUMRANGER_RELABEL_RESEGMENT } from '../subworkflows/local/xeniumranger_relabel_resegment/main' include { XENIUMRANGER_IMPORT_SEGMENTATION_REDEFINE_BUNDLE } from '../subworkflows/local/xeniumranger_import_segmentation_redefine_bundle/main' // spatialdata subworkflows -include { SPATIALDATA_WRITE_META_MERGE } from '../subworkflows/local/spatialdata_write_meta_merge/main' +include { SPATIALDATA_WRITE_META_MERGE } from '../subworkflows/local/spatialdata_write_meta_merge/main' // TODO qc layer subworkflows @@ -75,7 +72,7 @@ workflow SPATIALXE { ch_morphology_image = Channel.empty() ch_redefined_bundle = Channel.empty() ch_config = Channel.empty() - + ch_input = Channel.empty() /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ @@ -83,55 +80,97 @@ workflow SPATIALXE { ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ */ - // get sample, xenium bundle and image path - ch_bundle_path = ch_samplesheet.map { meta, bundle, _image -> - return [ meta, bundle ] + if ( workflow.profile.contains('test') ) { + + // get sample, xenium bundle and image path + ch_input_untar = ch_samplesheet.map { meta, bundle, _image -> + return [ meta, bundle ] + } + + // get testdata + UNTAR ( ch_input_untar ) + ch_versions = ch_versions.mix ( UNTAR.out.versions ) + + ch_untar_outs = UNTAR.out.untar.map { _meta, bundle -> + return [ bundle.toString() ] + } + + ch_samplesheet.combine(ch_untar_outs) + .map { meta, _url, image, test_bundle -> + return [meta, test_bundle, image] + } + .set { ch_input } + + ch_input.view() + + } else { + + // for all other profile runs + ch_input = ch_samplesheet } - // get all files in the xenium bundle in a channel - ch_bundle_files = ch_samplesheet.map - { meta, bundle_path, _image -> - def files = file("${bundle_path}/*").collect() - [meta, [files]] - } + // path to bundle input + ch_bundle_path = ch_input.map { meta, bundle, _image -> + return [ meta, bundle ] + } // get transcript.csv.gz from the xenium bundle - ch_transcripts_csv = ch_samplesheet.map { meta, bundle, _image -> - def transcripts_csv = file(bundle.replaceFirst(/\/$/, '') + "/transcripts.csv.gz") + ch_transcripts_csv = ch_input.map { meta, bundle, _image -> + def transcripts_csv = file ( + bundle.toString().replaceFirst(/\/$/, '') + "/transcripts.csv.gz", + checkIfExists: true + ) return [ meta, transcripts_csv ] } // get transcript.parquet from the xenium bundle - ch_transcripts_parquet = ch_samplesheet.map { meta, bundle, _image -> - def transcripts_parquet = file(bundle.replaceFirst(/\/$/, '') + "/transcripts.parquet") + ch_transcripts_parquet = ch_input.map { meta, bundle, _image -> + def transcripts_parquet = file ( + bundle.toString().replaceFirst(/\/$/, '') + "/transcripts.parquet", + checkIfExists: true + ) return [ meta, transcripts_parquet ] } // get morphology.ome.tif from the xenium bundle - ch_morphology_image = ch_samplesheet.map { meta, bundle, image -> - def morphology_img = image ? file(image) : file(bundle.replaceFirst(/\/$/, '') + "/morphology.ome.tif") + ch_morphology_image = ch_input.map { meta, bundle, image -> + def morphology_img = image ? file(image) : file ( + bundle.toString().replaceFirst(/\/$/, '') + "/morphology.ome.tif", + checkIfExists: true + ) return [ meta, morphology_img ] } // get baysor xenium config - ch_config = Channel.fromPath("${projectDir}/assets/config/xenium.toml", checkIfExists: true) + ch_config = Channel.fromPath ( + "${projectDir}/assets/config/xenium.toml", + checkIfExists: true + ) // get segmentation mask if provided with --segmentation_mask for the baysor method if ( params.segmentation_mask ) { - ch_segmentation_mask = Channel.fromPath(params.segmentation_mask, checkIfExists: true) + ch_segmentation_mask = Channel.fromPath ( + params.segmentation_mask, checkIfExists: true + ) } // get gene_panel.json if provided with --gene_panel, sets relabel_genes to true if (( params.gene_panel )) { params.relabel_genes = true - ch_gene_panel = Channel.fromPath(params.gene_panel, checkIfExists: true) + ch_gene_panel = Channel.fromPath ( + params.gene_panel, + checkIfExists: true + ) } else { // gene panel to use if only --relabel_genes is provided - ch_gene_panel = ch_samplesheet.map { meta, bundle, _image -> - def gene_panel = file(bundle.replaceFirst(/\/$/, '') + "/gene_panel.json") + ch_gene_panel = ch_input.map { meta, bundle, _image -> + def gene_panel = file ( + bundle.toString().replaceFirst(/\/$/, '') + "/gene_panel.json", + checkIfExists: true + ) return [ meta, gene_panel ] } } From 248b7b2d705098870b193a2c6a828d85d043e59f Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Mon, 19 May 2025 13:47:29 +0000 Subject: [PATCH 06/24] changed transcripts handling to parquet file from csv --- conf/modules.config | 4 ++ modules.json | 3 +- .../spatialconverter/parquet_to_csv/main.nf | 12 +++--- modules/nf-core/gunzip/gunzip.diff | 41 +++++++++++++++++++ modules/nf-core/gunzip/main.nf | 10 ++--- nextflow.config | 3 -- nextflow_schema.json | 15 +------ .../local/baysor_generate_preview/main.nf | 16 ++++---- .../local/baysor_generate_segfree/main.nf | 16 ++++---- .../main.nf | 22 ++++------ .../local/baysor_run_transcripts_csv/main.nf | 16 ++------ .../local/proseg_preset_proseg2baysor/main.nf | 17 ++++++-- workflows/spatialxe.nf | 26 ++++-------- 13 files changed, 105 insertions(+), 96 deletions(-) create mode 100644 modules/nf-core/gunzip/gunzip.diff diff --git a/conf/modules.config b/conf/modules.config index 254a9d1a..5f9e93fe 100644 --- a/conf/modules.config +++ b/conf/modules.config @@ -27,6 +27,10 @@ process { ] } + withName: GUNZIP { + ext.prefix = "transcripts.csv" + } + withName: XENIUMRANGER_RESEGMENT { publishDir = [ path: "${params.outdir}/xeniumranger/resegment", diff --git a/modules.json b/modules.json index 920f97ee..07119f43 100644 --- a/modules.json +++ b/modules.json @@ -14,7 +14,8 @@ "gunzip": { "branch": "master", "git_sha": "81880787133db07d9b4c1febd152c090eb8325dc", - "installed_by": ["modules"] + "installed_by": ["modules"], + "patch": "modules/nf-core/gunzip/gunzip.diff" }, "multiqc": { "branch": "master", diff --git a/modules/local/spatialconverter/parquet_to_csv/main.nf b/modules/local/spatialconverter/parquet_to_csv/main.nf index 8f59157e..fad62cd5 100644 --- a/modules/local/spatialconverter/parquet_to_csv/main.nf +++ b/modules/local/spatialconverter/parquet_to_csv/main.nf @@ -4,21 +4,21 @@ process PARQUET_TO_CSV { container "ghcr.io/scverse/spatialdata:spatialdata0.3.0_spatialdata-io0.1.7_spatialdata-plot0.2.9" - if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) { - error "PARQUET_TO_CSV module does not support Conda. Please use Docker / Singularity / Podman instead." - } - input: tuple val(meta), path(transcripts) output: - tuple val(meta), path("*.csv") , emit: transcripts_csv - path("versions.yml") , emit: versions + tuple val(meta), path("*.csv"), emit: transcripts_csv + path("versions.yml") , emit: versions when: task.ext.when == null || task.ext.when script: + if (workflow.profile.tokenize(',').intersect(['conda', 'mamba']).size() >= 1) { + error "PARQUET_TO_CSV module does not support Conda. Please use Docker / Singularity / Podman instead." + } + template 'parquet_to_csv.py' stub: diff --git a/modules/nf-core/gunzip/gunzip.diff b/modules/nf-core/gunzip/gunzip.diff new file mode 100644 index 00000000..2b8554ce --- /dev/null +++ b/modules/nf-core/gunzip/gunzip.diff @@ -0,0 +1,41 @@ +Changes in component 'nf-core/gunzip' +'modules/nf-core/gunzip/environment.yml' is unchanged +Changes in 'gunzip/main.nf': +--- modules/nf-core/gunzip/main.nf ++++ modules/nf-core/gunzip/main.nf +@@ -11,7 +11,7 @@ + tuple val(meta), path(archive) + + output: +- tuple val(meta), path("${gunzip}"), emit: gunzip ++ tuple val(meta), path("${archive}.gz"), emit: gunzip + path "versions.yml", emit: versions + + when: +@@ -19,16 +19,14 @@ + + script: + def args = task.ext.args ?: '' +- def extension = (archive.toString() - '.gz').tokenize('.')[-1] +- def name = archive.toString() - '.gz' - ".${extension}" +- def prefix = task.ext.prefix ?: name +- gunzip = prefix + ".${extension}" ++ def prefix = task.ext.prefix ++ def gunzip = "${prefix}" + ".gz" + """ + # Not calling gunzip itself because it creates files + # with the original group ownership rather than the + # default one for that user / the work directory + gzip \\ +- -cd \\ ++ -c \\ + ${args} \\ + ${archive} \\ + > ${gunzip} + +'modules/nf-core/gunzip/meta.yml' is unchanged +'modules/nf-core/gunzip/tests/main.nf.test' is unchanged +'modules/nf-core/gunzip/tests/main.nf.test.snap' is unchanged +'modules/nf-core/gunzip/tests/nextflow.config' is unchanged +'modules/nf-core/gunzip/tests/tags.yml' is unchanged +************************************************************ diff --git a/modules/nf-core/gunzip/main.nf b/modules/nf-core/gunzip/main.nf index 3ffc8e92..a41d0382 100644 --- a/modules/nf-core/gunzip/main.nf +++ b/modules/nf-core/gunzip/main.nf @@ -11,7 +11,7 @@ process GUNZIP { tuple val(meta), path(archive) output: - tuple val(meta), path("${gunzip}"), emit: gunzip + tuple val(meta), path("${archive}.gz"), emit: gunzip path "versions.yml", emit: versions when: @@ -19,16 +19,14 @@ process GUNZIP { script: def args = task.ext.args ?: '' - def extension = (archive.toString() - '.gz').tokenize('.')[-1] - def name = archive.toString() - '.gz' - ".${extension}" - def prefix = task.ext.prefix ?: name - gunzip = prefix + ".${extension}" + def prefix = task.ext.prefix + def gunzip = "${prefix}" + ".gz" """ # Not calling gunzip itself because it creates files # with the original group ownership rather than the # default one for that user / the work directory gzip \\ - -cd \\ + -c \\ ${args} \\ ${archive} \\ > ${gunzip} diff --git a/nextflow.config b/nextflow.config index 68e9f593..794eb3d6 100644 --- a/nextflow.config +++ b/nextflow.config @@ -51,9 +51,6 @@ params { features = null // Baysor specific - baysor_run_image = true // run baysor with image/seg-mask - baysor_run_transcripts = false // run baysor with transcripts.csv.gz - baysor_preview = false // generate preview with baysor preview cmd // MultiQC options multiqc_config = null diff --git a/nextflow_schema.json b/nextflow_schema.json index 3b4749ce..d8aa12f8 100644 --- a/nextflow_schema.json +++ b/nextflow_schema.json @@ -85,7 +85,7 @@ "properties": { "generate_preview": { "type": "boolean", - "description": "Whether to generate a preview of the dataset with the transcripts.csv.gz." + "description": "Whether to generate a preview of the dataset with the transcripts.parquet." }, "segmentation_refinement": { "type": "boolean", @@ -175,19 +175,6 @@ "features": { "type": "string", "description": "List of features to be passed to the ficture method. (eg: TP53,OCIAD1,BCAS3,SOX)" - }, - "baysor_run_image": { - "type": "boolean", - "default": true, - "description": "Whether to run bayor with image/segmentation-mask." - }, - "baysor_run_transcripts": { - "type": "boolean", - "description": "Whether to run baysor with transcripts.csv.gz." - }, - "baysor_preview": { - "type": "boolean", - "description": "Whether to create a preview of the dataset with transcripts.csv.gz." } } }, diff --git a/subworkflows/local/baysor_generate_preview/main.nf b/subworkflows/local/baysor_generate_preview/main.nf index 59f3b810..d3c6589d 100644 --- a/subworkflows/local/baysor_generate_preview/main.nf +++ b/subworkflows/local/baysor_generate_preview/main.nf @@ -2,16 +2,16 @@ // Run baysor create_dataset & preview // -include { GUNZIP } from '../../../modules/nf-core/gunzip/main' include { BAYSOR_PREVIEW } from '../../../modules/local/baysor/preview/main' include { BAYSOR_CREATE_DATASET } from '../../../modules/local/baysor/create_dataset/main' +include { PARQUET_TO_CSV } from '../../../modules/local/spatialconverter/parquet_to_csv/main' workflow BAYSOR_GENERATE_PREVIEW { take: - ch_transcripts_csv // channel: [ val(meta), ["path-to-transcripts.csv.gz"] ] - ch_config // channel: ["path-to-xenium.toml"] + ch_transcripts_parquet // channel: [ val(meta), ["path-to-transcripts.parquet"] ] + ch_config // channel: ["path-to-xenium.toml"] main: @@ -19,14 +19,12 @@ workflow BAYSOR_GENERATE_PREVIEW { ch_preview_html = Channel.empty() - // unzip transcripts.csv.gz - GUNZIP ( ch_transcripts_csv ) - ch_versions = ch_versions.mix ( GUNZIP.out.versions ) - - ch_unzipped_transcripts = GUNZIP.out.gunzip + // run parquet to csv + PARQUET_TO_CSV ( ch_transcripts_parquet ) + ch_versions = ch_versions.mix ( PARQUET_TO_CSV.out.versions ) // generate randomised sample data - BAYSOR_CREATE_DATASET ( ch_unzipped_transcripts, "0.3" ) + BAYSOR_CREATE_DATASET ( PARQUET_TO_CSV.out.transcripts_csv, "0.3" ) ch_versions = ch_versions.mix ( BAYSOR_CREATE_DATASET.out.versions ) // run baysor preview if param - generate_preview is true diff --git a/subworkflows/local/baysor_generate_segfree/main.nf b/subworkflows/local/baysor_generate_segfree/main.nf index c194341b..463470f8 100644 --- a/subworkflows/local/baysor_generate_segfree/main.nf +++ b/subworkflows/local/baysor_generate_segfree/main.nf @@ -10,7 +10,8 @@ workflow BAYSOR_GENERATE_SEGFREE { take: - ch_transcripts // channel: [ val(meta), ["transcripts.csv.gz"] ] + ch_transcripts_parquet // channel: [ val(meta), ["transcripts.parquet"] ] + ch_config main: @@ -18,19 +19,18 @@ workflow BAYSOR_GENERATE_SEGFREE { ch_ncvs = Channel.empty() - // unzip transcripts.csv.gz - GUNZIP ( ch_transcripts ) - ch_versions = ch_versions.mix ( GUNZIP.out.versions ) - // run baysor segfree BAYSOR_SEGFREE ( - GUNZIP.out.gunzip + ch_transcripts_parquet, + ch_config ) ch_versions = ch_versions.mix( BAYSOR_SEGFREE.out.versions ) + ch_ncvs = BAYSOR_SEGFREE.out.ncvs + emit: - ncvs = ch_ncvs + ncvs = ch_ncvs // channel: [ val(meta), ["ncvs.loom"] ] - versions = ch_versions // channel: [ versions.yml ] + versions = ch_versions // channel: [ versions.yml ] } diff --git a/subworkflows/local/baysor_run_prior_segmentation_mask/main.nf b/subworkflows/local/baysor_run_prior_segmentation_mask/main.nf index a241f21c..40a4924e 100644 --- a/subworkflows/local/baysor_run_prior_segmentation_mask/main.nf +++ b/subworkflows/local/baysor_run_prior_segmentation_mask/main.nf @@ -2,20 +2,18 @@ // Run baysor run & import-segmentation // -include { GUNZIP } from '../../../modules/nf-core/gunzip/main' -include { RESOLIFT } from '../../../modules/local/resolift/main' -include { BAYSOR_RUN as BAYSOR_RUN_IMAGE } from '../../../modules/local/baysor/run/main' -include { XENIUMRANGER_IMPORT_SEGMENTATION } from '../../../modules/nf-core/xeniumranger/import-segmentation/main' +include { BAYSOR_RUN as BAYSOR_RUN_IMAGE } from '../../../modules/local/baysor/run/main' +include { XENIUMRANGER_IMPORT_SEGMENTATION } from '../../../modules/nf-core/xeniumranger/import-segmentation/main' workflow BAYSOR_RUN_PRIOR_SEGMENTATION_MASK { take: - ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] - ch_transcripts_csv // channel: [ val(meta), ["path-to-transcripts.csv.gz"] ] - ch_segmentation_mask // channel: [ ["path-to-prior-segmentation-mask"] ] - ch_config // channel: ["path-to-xenium.toml"] + ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] + ch_transcripts_parquet // channel: [ val(meta), ["path-to-transcripts.parquet"] ] + ch_segmentation_mask // channel: [ ["path-to-prior-segmentation-mask"] ] + ch_config // channel: [ "path-to-xenium.toml" ] main: @@ -26,17 +24,11 @@ workflow BAYSOR_RUN_PRIOR_SEGMENTATION_MASK { ch_htmls = Channel.empty() ch_redefined_bundle = Channel.empty() - ch_unzipped_transcripts = Channel.empty() - // unzip transcripts.csv.gz - GUNZIP ( ch_transcripts_csv ) - ch_versions = ch_versions.mix ( GUNZIP.out.versions ) - - ch_unzipped_transcripts = GUNZIP.out.gunzip // run baysor with morphology.tiff BAYSOR_RUN_IMAGE ( - ch_unzipped_transcripts, + ch_transcripts_parquet, ch_segmentation_mask, ch_config, 30 diff --git a/subworkflows/local/baysor_run_transcripts_csv/main.nf b/subworkflows/local/baysor_run_transcripts_csv/main.nf index 10b067d8..e426c6f2 100644 --- a/subworkflows/local/baysor_run_transcripts_csv/main.nf +++ b/subworkflows/local/baysor_run_transcripts_csv/main.nf @@ -11,9 +11,9 @@ workflow BAYSOR_RUN_TRANSCRIPTS_CSV { take: - ch_bundle_path // channel: [ val(meta), ["xenium-bundle"] ] - ch_transcripts_csv // channel: [ val(meta), ["transcripts.csv.gz"] ] - ch_config // channel: ["path-to-xenium.toml"] + ch_bundle_path // channel: [ val(meta), ["xenium-bundle"] ] + ch_transcripts_parquet // channel: [ val(meta), ["transcripts.csv.parquet"] ] + ch_config // channel: ["path-to-xenium.toml"] main: @@ -24,18 +24,10 @@ workflow BAYSOR_RUN_TRANSCRIPTS_CSV { ch_htmls = Channel.empty() ch_redefined_bundle = Channel.empty() - ch_unzipped_transcripts = Channel.empty() - - - // unzip transcripts.csv.gz - GUNZIP ( ch_transcripts_csv ) - ch_versions = ch_versions.mix ( GUNZIP.out.versions ) - - ch_unzipped_transcripts = GUNZIP.out.gunzip // run baysor with transcripts.csv BAYSOR_RUN_TRANSCRIPTS ( - ch_unzipped_transcripts, + ch_transcripts_parquet, [], ch_config, 30 diff --git a/subworkflows/local/proseg_preset_proseg2baysor/main.nf b/subworkflows/local/proseg_preset_proseg2baysor/main.nf index 20c90f64..53db2bd0 100644 --- a/subworkflows/local/proseg_preset_proseg2baysor/main.nf +++ b/subworkflows/local/proseg_preset_proseg2baysor/main.nf @@ -2,26 +2,35 @@ // Runs proseg for the xenium format and proseg2baysor to generate cell ploygons // +include { GUNZIP } from '../../../modules/nf-core/gunzip/main' include { PROSEG } from '../../../modules/local/proseg/preset/main' include { PROSEG2BAYSOR } from '../../../modules/local/proseg/proseg2baysor/main' +include { PARQUET_TO_CSV } from '../../../modules/local/spatialconverter/parquet_to_csv/main' include { XENIUMRANGER_IMPORT_SEGMENTATION } from '../../../modules/nf-core/xeniumranger/import-segmentation/main' workflow PROSEG_PRESET_PROSEG2BAYSOR { take: - ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] - ch_transcripts_csv // channel: [ val(meta), [ "transcripts.csv.gz" ] ] + ch_bundle_path // channel: [ val(meta), ["path-to-xenium-bundle"] ] + ch_transcripts_parquet // channel: [ val(meta), [ "transcripts.parquet" ] ] main: ch_versions = Channel.empty() + // run parquet-to-csv + PARQUET_TO_CSV ( ch_transcripts_parquet ) + ch_versions = ch_versions.mix( PARQUET_TO_CSV.out.versions ) + + // run gzip to create `transcripts.csv.gz` + GUNZIP ( PARQUET_TO_CSV.out.transcripts_csv ) + ch_versions = ch_versions.mix( GUNZIP.out.versions ) + // run proseg with the xenium format - PROSEG ( ch_transcripts_csv ) + PROSEG ( GUNZIP.out.gunzip ) ch_versions = ch_versions.mix( PROSEG.out.versions ) - // run proseg-to-baysor on the data generated with the proseg run PROSEG2BAYSOR ( PROSEG.out.cell_polygons_2d, PROSEG.out.transcript_metadata ) ch_versions = ch_versions.mix( PROSEG2BAYSOR.out.versions ) diff --git a/workflows/spatialxe.nf b/workflows/spatialxe.nf index 5a1e7a50..afddfe6b 100644 --- a/workflows/spatialxe.nf +++ b/workflows/spatialxe.nf @@ -67,7 +67,6 @@ workflow SPATIALXE { ch_bundle_path = Channel.empty() ch_raw_bundle = Channel.empty() ch_gene_panel = Channel.empty() - ch_transcripts_csv = Channel.empty() ch_transcripts_parquet = Channel.empty() ch_morphology_image = Channel.empty() ch_redefined_bundle = Channel.empty() @@ -114,15 +113,6 @@ workflow SPATIALXE { return [ meta, bundle ] } - // get transcript.csv.gz from the xenium bundle - ch_transcripts_csv = ch_input.map { meta, bundle, _image -> - def transcripts_csv = file ( - bundle.toString().replaceFirst(/\/$/, '') + "/transcripts.csv.gz", - checkIfExists: true - ) - return [ meta, transcripts_csv ] - } - // get transcript.parquet from the xenium bundle ch_transcripts_parquet = ch_input.map { meta, bundle, _image -> def transcripts_parquet = file ( @@ -203,7 +193,7 @@ workflow SPATIALXE { if ( params.generate_preview && params.mode == 'coordinate' ) { BAYSOR_GENERATE_PREVIEW ( - ch_transcripts_csv, + ch_transcripts_parquet, ch_config ) log.info "Preview generated at ${params.outdir}" @@ -246,7 +236,7 @@ workflow SPATIALXE { BAYSOR_RUN_PRIOR_SEGMENTATION_MASK ( ch_bundle_path, - ch_transcripts_csv, + ch_transcripts_parquet, ch_segmentation_mask, ch_config ) @@ -278,7 +268,7 @@ workflow SPATIALXE { PROSEG_PRESET_PROSEG2BAYSOR ( ch_bundle_path, - ch_transcripts_csv + ch_transcripts_parquet ) ch_redefined_bundle = PROSEG_PRESET_PROSEG2BAYSOR.out.redefined_bundle @@ -287,18 +277,18 @@ workflow SPATIALXE { // check it the provided method is part of the methods list if ( params.segmentation in params.transcript_seg_methods ) { - // run proseg with transcripts.csv.gz + // run proseg with transcripts.parquet if ( params.segmentation == 'proseg') { PROSEG_PRESET_PROSEG2BAYSOR ( ch_bundle_path, - ch_transcripts_csv + ch_transcripts_parquet ) ch_redefined_bundle = PROSEG_PRESET_PROSEG2BAYSOR.out.redefined_bundle } - // run segger with transcripts.csv.gz + // run segger with transcripts.parquet if ( params.segmentation == 'segger' ) { SEGGER_CREATE_TRAIN_PREDICT ( @@ -308,12 +298,12 @@ workflow SPATIALXE { } - // run baysor with transcripts.csv.gz + // run baysor with transcripts.parquet if ( params.segmentation == 'baysor' ) { BAYSOR_RUN_TRANSCRIPTS_CSV ( ch_bundle_path, - ch_transcripts_csv, + ch_transcripts_parquet, ch_config ) ch_redefined_bundle = BAYSOR_RUN_TRANSCRIPTS_CSV.out.redefined_bundle From 0703cd23f185e3649abc0049999c735bad75c02e Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Mon, 19 May 2025 14:00:25 +0000 Subject: [PATCH 07/24] cpu num fix for cellpose --- conf/test.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test.config b/conf/test.config index 8e39243c..57c68300 100644 --- a/conf/test.config +++ b/conf/test.config @@ -13,7 +13,7 @@ process { resourceLimits = [ - cpus: 8, + cpus: 4, memory: '16.GB', time: '1.h' ] From a487b604bf689fd0e9a4c2d3bc4832832e335d0e Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Mon, 19 May 2025 14:04:39 +0000 Subject: [PATCH 08/24] memory fix for cellpose --- conf/test.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test.config b/conf/test.config index 57c68300..3c2aa97a 100644 --- a/conf/test.config +++ b/conf/test.config @@ -14,7 +14,7 @@ process { resourceLimits = [ cpus: 4, - memory: '16.GB', + memory: '15.GB', time: '1.h' ] } From 81d196007cb027a673ac067e2c3e21bf982fbad7 Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Mon, 19 May 2025 15:42:56 +0000 Subject: [PATCH 09/24] fix for imp-seg pipestance errors --- README.md | 2 +- conf/test.config | 4 ++++ subworkflows/local/xeniumranger_relabel_resegment/main.nf | 1 - workflows/spatialxe.nf | 2 -- 4 files changed, 5 insertions(+), 4 deletions(-) diff --git a/README.md b/README.md index 0b53aba4..8b111e60 100644 --- a/README.md +++ b/README.md @@ -61,7 +61,7 @@ nextflow run nf-core/spatialxe \ ## Run coordinate-based segmentation mode
-`PROSEG -> BAYSOR -> XR-IMPORT_SEGMENTATION -> SPATIALDATA -> QC` +`PROSEG -> PROSEG2BAYSOR -> XR-IMPORT_SEGMENTATION -> SPATIALDATA -> QC` ```bash nextflow run nf-core/spatialxe \ diff --git a/conf/test.config b/conf/test.config index 3c2aa97a..a4169fdf 100644 --- a/conf/test.config +++ b/conf/test.config @@ -17,6 +17,10 @@ process { memory: '15.GB', time: '1.h' ] + + withName: UNTAR { + ext.prefix = "test_bundle" + } } params { diff --git a/subworkflows/local/xeniumranger_relabel_resegment/main.nf b/subworkflows/local/xeniumranger_relabel_resegment/main.nf index 99075ccb..1151e69a 100644 --- a/subworkflows/local/xeniumranger_relabel_resegment/main.nf +++ b/subworkflows/local/xeniumranger_relabel_resegment/main.nf @@ -16,7 +16,6 @@ workflow XENIUMRANGER_RELABEL_RESEGMENT { ch_versions = Channel.empty() - ch_bundle_path.view() XENIUMRANGER_RELABEL ( ch_bundle_path, ch_gene_panel diff --git a/workflows/spatialxe.nf b/workflows/spatialxe.nf index afddfe6b..478ca3be 100644 --- a/workflows/spatialxe.nf +++ b/workflows/spatialxe.nf @@ -100,8 +100,6 @@ workflow SPATIALXE { } .set { ch_input } - ch_input.view() - } else { // for all other profile runs From cd6ba069720707b0c11697529502c2f0780f88cb Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Mon, 19 May 2025 17:12:12 +0000 Subject: [PATCH 10/24] fixed time for CI test with reduced resources --- conf/test.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test.config b/conf/test.config index a4169fdf..4a5e3c94 100644 --- a/conf/test.config +++ b/conf/test.config @@ -15,7 +15,7 @@ process { resourceLimits = [ cpus: 4, memory: '15.GB', - time: '1.h' + time: '2.h' ] withName: UNTAR { From 50431e803f6cde6710dba160faecf5e49540aebf Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Mon, 19 May 2025 23:28:58 +0000 Subject: [PATCH 11/24] validator function for input xenium bundle --- .../utils_nfcore_spatialxe_pipeline/main.nf | 100 ++++++++++++++++-- workflows/spatialxe.nf | 10 +- 2 files changed, 95 insertions(+), 15 deletions(-) diff --git a/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf b/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf index bd13cea6..814651d6 100644 --- a/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf @@ -67,22 +67,36 @@ workflow PIPELINE_INITIALISATION { // Custom validation for pipeline parameters // validateInputParameters() + log.info "INFO ✅ Input params validated" // // Create channel from input file provided through params.input // - Channel + try { + + Channel .fromList(samplesheetToList(input, "${projectDir}/assets/schema_input.json")) .map { meta, bundle, image -> return [ [id: meta.id], bundle, image ] } .set { ch_samplesheet } + log.info "INFO ✅ Samplesheet validated" + + } catch (Exception e) { + + log.error "❌ Samplesheet validation failed: ${e.message}" + exit 1 + } + // // Check and validate xenium bundle // - validateXeniumBundle() + if ( !workflow.profile.contains('test')) { + validateXeniumBundle(ch_samplesheet) + } + emit: @@ -136,7 +150,7 @@ workflow PIPELINE_COMPLETION { } workflow.onError { - log.error "Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting" + log.error "❌ Pipeline failed. Please refer to troubleshooting docs: https://nf-co.re/docs/usage/troubleshooting" } } @@ -153,35 +167,38 @@ def validateInputParameters() { // check if the segmentation method provided is valid for a mode if ( params.mode == 'image' && params.segmentation ) { if ( !params.image_seg_methods.contains(params.segmentation) ) { - error "Error: Invalid segmentation method: ${params.segmentation} provided for the `image` based mode. Options: ${params.image_seg_methods}" + log.error "❌ Error: Invalid segmentation method: ${params.segmentation} provided for the `image` based mode. Options: ${params.image_seg_methods}" + exit 1 } } if ( params.mode == 'coordinate' && params.segmentation ) { if ( !params.transcript_seg_methods.contains(params.segmentation) ) { - error "Error: Invalid segmentation method: `${params.segmentation}` provided for the `coordinate` based mode. Options: ${params.transcript_seg_methods}" + log.error "❌ Error: Invalid segmentation method: `${params.segmentation}` provided for the `coordinate` based mode. Options: ${params.transcript_seg_methods}" + exit 1 } } // check if --relabel_genes is true but --gene_panel is not provided if ( params.relabel_genes && !params.gene_panel ) { - log.warn "Relabel genes is enabled, but gene panel is not provided with the `--gene_panel`. Using `gene_panel.json` in the xenium bundle." + log.warn "⚠️ Relabel genes is enabled, but gene panel is not provided with the `--gene_panel`. Using `gene_panel.json` in the xenium bundle." } // check if --relabel_genes is true but --gene_panel is not provided if ( params.gene_panel && !params.relabel_genes ) { - log.warn "Gene panel provided, but relabel genes is disabled. Using `gene_panel.json` only to generate metadata." + log.warn "⚠️ Gene panel provided, but relabel genes is disabled. Using `gene_panel.json` only to generate metadata." } // check if segmentation method is xeniumranger and nucleus_ony_segmentation is enabled if ( params.segmentation == 'xeniumranger' && !params.nucleus_segmentation_only ) { - log.warn "Nucleus segmentation is disabled. Running xeniumranger resegment module to redefine xenium bundle without nucleus segmentation." - log.warn "Use --nucleus_segmentation_only to enable nucleus segmentation to redefine xenium bundle with import-segmentation module." + log.warn "⚠️ Nucleus segmentation is disabled. Running xeniumranger resegment module to redefine xenium bundle without nucleus segmentation." + log.warn "⚠️ Use --nucleus_segmentation_only to enable nucleus segmentation to redefine xenium bundle with import-segmentation module." } if ( params.mode == 'image' && params.segmentation == 'baysor' ) { if ( !params.segmentation_mask ) { - error "Error: Missing path to segmentation mask. Image-based segmentation with the `baysor` method requires a segmentation mask with the `--segmentation_mask` option." + log.error "❌ Error: Missing path to segmentation mask. Image-based segmentation with the `baysor` method requires a segmentation mask with the `--segmentation_mask` option." + exit 1 } } @@ -190,8 +207,69 @@ def validateInputParameters() { // // Check and validate xenium bundle // -def validateXeniumBundle() { +def validateXeniumBundle(ch_samplesheet) { + + // define xenium bundle directory structure + def xenium_bundle = [ + "analysis.tar.gz", + "analysis.zarr.zip", + "analysis_summary.html", + "cell_boundaries.csv.gz", + "cell_boundaries.parquet", + "cell_feature_matrix.h5", + "cell_feature_matrix.tar.gz", + "cell_feature_matrix.zarr.zip", + "cells.csv.gz", + "cells.parquet", + "cells.zarr.zip", + "experiment.xenium", + "gene_panel.json", + "metrics_summary.csv", + "morphology.ome.tif", + "morphology_focus/", + "nucleus_boundaries.csv.gz", + "nucleus_boundaries.parquet", + "transcripts.parquet", + "transcripts.zarr.zip" + ] + + // get bundle path + def ch_bundle_path = ch_samplesheet.map { + _meta, bundle, _image -> + def bundle_path = file ( + bundle.toString().replaceFirst(/\/$/, ''), + ) + return bundle_path + } + + // check if the path exists + if ( !ch_bundle_path.map { it.exists() } ) { + error "❌ Error: Xenium bundle path not found. Check if the path provided in the samplesheet exists." + exit 1 + } + // if the path exists, check for the presence of xenium files + if ( ch_bundle_path.map { it.exists() } ) { + + ch_bundle_path.map { path -> + def missing_files = [] + + def allExist = xenium_bundle.every { filename -> + def fullPath = file("${path}/${filename}") + if (!fullPath.exists()) { + missing_files.add(filename) + return false + } + return true + } + + if (!allExist) { + log.error "❌ Missing file(s) at bundle path provided in the samplesheet: ${missing_files}" + exit 1 + } + } + } + log.info "INFO ✅ Xenium bundle validated" } // diff --git a/workflows/spatialxe.nf b/workflows/spatialxe.nf index 478ca3be..9d03e2fd 100644 --- a/workflows/spatialxe.nf +++ b/workflows/spatialxe.nf @@ -62,16 +62,18 @@ workflow SPATIALXE { */ ch_versions = Channel.empty() - ch_multiqc_files = Channel.empty() + + ch_input = Channel.empty() ch_bundle = Channel.empty() + ch_config = Channel.empty() ch_bundle_path = Channel.empty() ch_raw_bundle = Channel.empty() ch_gene_panel = Channel.empty() - ch_transcripts_parquet = Channel.empty() + ch_multiqc_files = Channel.empty() ch_morphology_image = Channel.empty() ch_redefined_bundle = Channel.empty() - ch_config = Channel.empty() - ch_input = Channel.empty() + ch_transcripts_parquet = Channel.empty() + /* ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ From 2fd92b8c142a3c0d4b875ee4786deac43d498bde Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Tue, 20 May 2025 12:06:43 +0000 Subject: [PATCH 12/24] xenium bundle validator, minor changes for ci --- conf/test.config | 2 +- .../main.nf | 2 +- workflows/spatialxe.nf | 12 ++++++------ 3 files changed, 8 insertions(+), 8 deletions(-) rename subworkflows/local/{baysor_run_transcripts_csv => baysor_run_transcripts_parquet}/main.nf (98%) diff --git a/conf/test.config b/conf/test.config index 4a5e3c94..9e04c05a 100644 --- a/conf/test.config +++ b/conf/test.config @@ -15,7 +15,7 @@ process { resourceLimits = [ cpus: 4, memory: '15.GB', - time: '2.h' + time: '3.h' ] withName: UNTAR { diff --git a/subworkflows/local/baysor_run_transcripts_csv/main.nf b/subworkflows/local/baysor_run_transcripts_parquet/main.nf similarity index 98% rename from subworkflows/local/baysor_run_transcripts_csv/main.nf rename to subworkflows/local/baysor_run_transcripts_parquet/main.nf index e426c6f2..ce8b09ed 100644 --- a/subworkflows/local/baysor_run_transcripts_csv/main.nf +++ b/subworkflows/local/baysor_run_transcripts_parquet/main.nf @@ -7,7 +7,7 @@ include { BAYSOR_RUN as BAYSOR_RUN_TRANSCRIPTS } from '../../../modules/local/ba include { XENIUMRANGER_IMPORT_SEGMENTATION } from '../../../modules/nf-core/xeniumranger/import-segmentation/main' -workflow BAYSOR_RUN_TRANSCRIPTS_CSV { +workflow BAYSOR_RUN_TRANSCRIPTS_PARQUET { take: diff --git a/workflows/spatialxe.nf b/workflows/spatialxe.nf index 9d03e2fd..282936b7 100644 --- a/workflows/spatialxe.nf +++ b/workflows/spatialxe.nf @@ -17,10 +17,10 @@ include { paramsSummaryMap } from 'plugin/nf-schema' include { UNTAR } from '../modules/nf-core/untar/main' // coordinate-based segmentation subworklfows -include { SEGGER_CREATE_TRAIN_PREDICT } from '../subworkflows/local/segger_create_train_predict/main' -include { PROSEG_PRESET_PROSEG2BAYSOR } from '../subworkflows/local/proseg_preset_proseg2baysor/main' -include { BAYSOR_GENERATE_PREVIEW } from '../subworkflows/local/baysor_generate_preview/main' -include { BAYSOR_RUN_TRANSCRIPTS_CSV } from '../subworkflows/local/baysor_run_transcripts_csv/main' +include { SEGGER_CREATE_TRAIN_PREDICT } from '../subworkflows/local/segger_create_train_predict/main' +include { PROSEG_PRESET_PROSEG2BAYSOR } from '../subworkflows/local/proseg_preset_proseg2baysor/main' +include { BAYSOR_GENERATE_PREVIEW } from '../subworkflows/local/baysor_generate_preview/main' +include { BAYSOR_RUN_TRANSCRIPTS_PARQUET } from '../subworkflows/local/baysor_run_transcripts_csv/main' // image-based segmentation subworklfows include { BAYSOR_RUN_PRIOR_SEGMENTATION_MASK } from '../subworkflows/local/baysor_run_prior_segmentation_mask/main' @@ -301,12 +301,12 @@ workflow SPATIALXE { // run baysor with transcripts.parquet if ( params.segmentation == 'baysor' ) { - BAYSOR_RUN_TRANSCRIPTS_CSV ( + BAYSOR_RUN_TRANSCRIPTS_PARQUET ( ch_bundle_path, ch_transcripts_parquet, ch_config ) - ch_redefined_bundle = BAYSOR_RUN_TRANSCRIPTS_CSV.out.redefined_bundle + ch_redefined_bundle = BAYSOR_RUN_TRANSCRIPTS_PARQUET.out.redefined_bundle } } From 5389b8046c355d24b0e9c1cfddcd4dd4b7f247d1 Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Tue, 20 May 2025 12:07:09 +0000 Subject: [PATCH 13/24] xenium bundle validator, minor changes for ci --- subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf b/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf index 814651d6..ccf14fa6 100644 --- a/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf +++ b/subworkflows/local/utils_nfcore_spatialxe_pipeline/main.nf @@ -67,7 +67,7 @@ workflow PIPELINE_INITIALISATION { // Custom validation for pipeline parameters // validateInputParameters() - log.info "INFO ✅ Input params validated" + log.info "INFO Input params validated ✅ " // // Create channel from input file provided through params.input @@ -81,7 +81,7 @@ workflow PIPELINE_INITIALISATION { } .set { ch_samplesheet } - log.info "INFO ✅ Samplesheet validated" + log.info "INFO Samplesheet validated ✅ " } catch (Exception e) { @@ -269,7 +269,7 @@ def validateXeniumBundle(ch_samplesheet) { } } } - log.info "INFO ✅ Xenium bundle validated" + log.info "INFO Xenium bundle validated ✅ " } // From 261a56e94694452fc05a77584dbc237a687b5709 Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Tue, 20 May 2025 12:14:17 +0000 Subject: [PATCH 14/24] xenium bundle validator, minor changes for ci --- workflows/spatialxe.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/workflows/spatialxe.nf b/workflows/spatialxe.nf index 282936b7..aa029e30 100644 --- a/workflows/spatialxe.nf +++ b/workflows/spatialxe.nf @@ -20,7 +20,7 @@ include { UNTAR } from '../modules/nf-core/untar/main' include { SEGGER_CREATE_TRAIN_PREDICT } from '../subworkflows/local/segger_create_train_predict/main' include { PROSEG_PRESET_PROSEG2BAYSOR } from '../subworkflows/local/proseg_preset_proseg2baysor/main' include { BAYSOR_GENERATE_PREVIEW } from '../subworkflows/local/baysor_generate_preview/main' -include { BAYSOR_RUN_TRANSCRIPTS_PARQUET } from '../subworkflows/local/baysor_run_transcripts_csv/main' +include { BAYSOR_RUN_TRANSCRIPTS_PARQUET } from '../subworkflows/local/baysor_run_transcripts_parquet/main' // image-based segmentation subworklfows include { BAYSOR_RUN_PRIOR_SEGMENTATION_MASK } from '../subworkflows/local/baysor_run_prior_segmentation_mask/main' From cd984ac47225f28174da4858b7414b7a6a5ca143 Mon Sep 17 00:00:00 2001 From: Sameesh Kher Date: Tue, 20 May 2025 20:13:50 +0000 Subject: [PATCH 15/24] time limit changes for ci --- conf/test.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/test.config b/conf/test.config index 9e04c05a..bca0a264 100644 --- a/conf/test.config +++ b/conf/test.config @@ -15,7 +15,7 @@ process { resourceLimits = [ cpus: 4, memory: '15.GB', - time: '3.h' + time: '6.h' ] withName: UNTAR { From 4cdb9809a30fb222ec4b1173e5dae17d20ce6afc Mon Sep 17 00:00:00 2001 From: khersameesh24 Date: Wed, 21 May 2025 02:57:05 +0100 Subject: [PATCH 16/24] added updated metromap for spatialxe --- README.md | 2 -- docs/images/spatialxe-metromap.png | Bin 1606748 -> 581307 bytes docs/images/spatialxe-metromap.svg | 5 +---- 3 files changed, 1 insertion(+), 6 deletions(-) diff --git a/README.md b/README.md index 8b111e60..5ca2c86e 100644 --- a/README.md +++ b/README.md @@ -26,8 +26,6 @@ -1. Present QC for raw reads ([`MultiQC`](http://multiqc.info/)) - ## Usage On release, automated continuous integration tests run the pipeline on a full-sized dataset on the AWS cloud infrastructure. This ensures that the pipeline runs on AWS, has sensible resource allocation defaults set to run on real-world datasets, and permits the persistent storage of results to benchmark between pipeline releases and other analysis sources. 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