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10x Genomics
Chromium Single Cell Gene Expression

Pipestance Structure

The pipeline output directory, described in Understanding Output, contains all of the data produced by one invocation of a pipeline (a pipestance) as well as rich metadata describing the characteristics of each stage. This directory contains a specific structure that is used by the Martian pipeline framework to track the state of the pipeline as execution proceeds.

Pipeline Structure

Cell Ranger's notion of a pipeline is very flexible in that a pipeline can be composed of stages that run stage code or sub-pipelines that may themselves contain stages or sub-pipelines.

Cell Ranger pipelines follow the convention that stages are named with verbs (e.g., ALIGN_READS, MARK_DUPLICATES, FILTER_BARCODES) and sub-pipelines are named with nouns and prefixed with an underscore (e.g., _BCSORTER). Each stage runs in its own directory bearing its name, and each stage's directory is contained within its parent pipeline's directory.

For example, the cellranger mkfastq pipeline has the following process graph:

where

Directory Structure

Every pipestance operates wholly inside of its pipeline output directory. When the pipestance completes, this pipestance output directory contains three outputs: metadata files, the pipestance output file directory, and the top-level pipeline stage directory.

The top-level pipeline stage directory is a stage directory that contains any number of child stage directories as well as one stage output directory for each fork run by that stage. There are four possible top-level pipeline stages:

All of the Cell Ranger pipelines only contain single-fork stages, so there will only ever be a fork0 stage output directory within each stage directory. Chunk output directories are a subset of stage output directories that additionally contain runtime information specific to the job or process being run by that chunk (e.g., a process ID or cluster job ID).

For example, the cellranger mkfastq pipeline's pipeline output directory contains the following directory structure:

_logMetadata file
outs/Pipestance output file directory
MAKE_FASTQS_CS/Top-level pipeline stage directory
MAKE_FASTQS_CS/fork0/Stage output directory
MAKE_FASTQS_CS/fork0/files/Stage output files
MAKE_FASTQS_CS/MAKE_FASTQS/Stage directory
MAKE_FASTQS_CS/MAKE_FASTQS/fork0/Stage output directory
MAKE_FASTQS_CS/MAKE_FASTQS/fork0/files/Stage output files
MAKE_FASTQS_CS/MAKE_FASTQS/BCL2FASTQ_WITH_SAMPLESHEET/Stage directory
MAKE_FASTQS_CS/MAKE_FASTQS/BCL2FASTQ_WITH_SAMPLESHEET/fork0/Stage output directory
MAKE_FASTQS_CS/MAKE_FASTQS/BCL2FASTQ_WITH_SAMPLESHEET/fork0/chnk0/Chunk output directory

Commonly Generated Metadata

The metadata contained in the pipeline output directory includes

File Name Description
_finalstateMetadata cache that is populated when a pipestance completes to minimize re-aggregation of metadata
_invocationThe MRO call used to invoke this pipestance
_logThe log messages that are reported to your terminal window when running cellranger commands
_mrosourceThe entire MRO describing the pipeline with all @include statements dereferenced
_perfDetailed runtime performance data for every stage in the pipestance
_timestampThe start and finish time for this pipestance
_vdrkillA list of all of the volatile data (temporary files) removed during pipeline execution as well as total number of files and bytes deleted
_versionsVersions of the components used by the pipeline

Stage directories contain stage output directories, stage output files, and the stage directories of any child stages or pipelines.

Stage output directories typically contain:

File Name Contents
files/Directory containing any files created by this stage that were not considered volatile (temporary)
split/A special stage output directory for the step that divided this stage's input into parallel chunks
chnkN/A chunk output directory for the Nth parallel chunk executed
join/A special stage output directory for the step that recombined this stage's parallel output chunks into a single output dataset again
_completeA file that, when present, signifies that this stage has successfully completed
_errorsA file that, when present, signifies that this stage failed. Contains the errors that resulted in stage failure.
_invocationThe MRO call used to execute this stage by the Martian framework
_outsThe output files generated by this stage
_vdrkillA list of all of the volatile data (temporary files) removed during pipeline execution as well as total number of files and bytes deleted

Chunk output directories are a subset of stage output directories that, in addition to the aforementioned stage output, may contain:

File Name Contents
_argsThe arguments passed to the stage's stage code
_jobinfoMetadata describing the stage's execution, including performance metrics, job manager jobid and jobname, and process ID
_jobscriptThe script submitted to the cluster job manager (cluster mode-only)
_stdoutAny stage code output that was printed to the stdout stream
_stderrAny stage code output that was printed to the stderr stream

These metadata files should be treated as read-only, and altering the contents of metadata files is not recommended.

Navigating Pipestances

Pipestance output directories can demonstrate very complicated structures, and re-attaching the Cell Ranger UI is the easiest way to quickly navigate to a pipeline stage of interest and examine its metadata. In the absence of being able to access the UI, the standard find command can quickly return high-level information about a pipestance.

For example, to find the stages that resulted in the overall failure of a pipestance whose output directory is sample345/,

$ find sample345/ -name _errors
sample345/SC_RNA_COUNTER_CS/SC_RNA_COUNTER/SUMMARIZE_REPORTS/fork0/chnk0/_errors

This tells us that the failed stage was SUMMARIZE_REPORTS.

It can be helpful to view all _errors files' contents at once by piping to xargs cat:

$ find sample345/ -name _errors | xargs cat
 
Traceback (most recent call last):
  File "/home/jdoe/cellranger-3.1.0/cellranger-cs/3.1.0/mro/stages/cellranger/summarize_reports/__init__.py", line 62, in main
    filtered_gene_bc_matrices=args.filtered_gene_bc_matrices)
  File "/home/jdoe/cellranger-3.1.0/cellranger-cs/3.1.0/lib/python/cellranger/webshim/common.py", line 625, in build_web_summary_html
    filtered_gene_bc_matrices=filtered_gene_bc_matrices)
  File "/home/jdoe/cellranger-3.1.0/cellranger-cs/3.1.0/lib/python/cellranger/webshim/common.py", line 614, in build_web_summary_json
    filtered_gene_bc_matrices=filtered_gene_bc_matrices)
  File "/home/jdoe/cellranger-3.1.0/cellranger-cs/3.1.0/lib/python/cellranger/webshim/common.py", line 579, in build_charts
    plot_data = plot_preprocess_func(sample_properties, filtered_matrices, gene_index)
  File "/home/jdoe/cellranger-3.1.0/cellranger-cs/3.1.0/lib/python/cellranger/webshim/common.py", line 319, in plot_preprocess
    silhouette_score = metrics.silhouette_score(kmeans_matrix, clusters, metric='cosine')
  File "/home/jdoe/cellranger-3.1.0/anaconda-cs/2.2.0/lib/python2.7/site-packages/sklearn/metrics/cluster/unsupervised.py", line 84, in silhouette_score
    "and less than n_samples - 1" % n_labels)
ValueError: Number of labels is 5 but should be more than 2 and less than n_samples - 1

In the above case, the error is an unhandled exception whose cause is not obvious; these sorts of failures should be reported to the 10x software support team for assistance with diagnosis.

Stages whose stage code run external binaries (for example, the ALIGN_READS stage which runs STAR) often generate output to their stdout and stderr streams. These messages are captured in the _stdout and _stderr metadata files within the chunk output directories, and combining find and xargs cat to examine their contents can also assist with troubleshooting.