From 9af665b5ef1cf139becda511600cb31694959320 Mon Sep 17 00:00:00 2001
From: Adam Freedman <adamfreedman@fas.harvard.edu>
Date: Wed, 10 Jun 2026 15:34:40 -0400
Subject: [PATCH 1/6] Allow full workflow test to run without cannon profile

---
 .github/workflows/full-workflow.yml |  6 ++++++
 tests/run_test_workflow.sh          | 14 +++++++++++---
 2 files changed, 17 insertions(+), 3 deletions(-)

diff --git a/.github/workflows/full-workflow.yml b/.github/workflows/full-workflow.yml
index a9a7360..34c8e95 100644
--- a/.github/workflows/full-workflow.yml
+++ b/.github/workflows/full-workflow.yml
@@ -21,6 +21,12 @@ jobs:
     env:
       SNAKEMAKE_CONDA_FRONTEND: mamba
       GITHUB_PAT: ${{ github.token }}
+      TEST_WORKFLOW_PROFILE_ARGS: >-
+        --profile none
+        --workflow-profile none
+        --executor local
+        --cores 2
+      TEST_WORKFLOW_JOBS: "2"
       TEST_WORKFLOW_SNAKEMAKE_ARGS: >-
         --resources mem_mb=8192
         --set-resources
diff --git a/tests/run_test_workflow.sh b/tests/run_test_workflow.sh
index 316a5cc..07207eb 100755
--- a/tests/run_test_workflow.sh
+++ b/tests/run_test_workflow.sh
@@ -13,6 +13,7 @@ set -euo pipefail
 usage() {
     echo "Usage: $0 [--conda-prefix PATH] [additional snakemake args...]"
     echo "Set TEST_WORKFLOW_SNAKEMAKE_ARGS to append whitespace-delimited Snakemake args before command-line extras."
+    echo "Set TEST_WORKFLOW_PROFILE_ARGS to replace the default Slurm/cannon profile args."
 }
 
 script_dir=$(cd "$(dirname "${BASH_SOURCE[0]}")" && pwd)
@@ -48,13 +49,18 @@ if ! command -v snakemake >/dev/null 2>&1; then
     exit 127
 fi
 
+profile_args=(--workflow-profile profiles/slurm --profile cannon)
+if [[ -n "${TEST_WORKFLOW_PROFILE_ARGS:-}" ]]; then
+    # Intended for simple Snakemake profile/executor args.
+    read -r -a profile_args <<< "${TEST_WORKFLOW_PROFILE_ARGS}"
+fi
+
 common_args=(
     --snakefile workflow/Snakefile
     --configfile config/config.yaml
     --config sampleTable=testdata/samplesheet_test.tsv resultsDir=testdata/results
     --use-conda
-    --workflow-profile profiles/slurm
-    --profile cannon
+    "${profile_args[@]}"
 )
 
 conda_prefix_args=()
@@ -62,6 +68,8 @@ if [[ -n "${snakemake_conda_prefix}" ]]; then
     conda_prefix_args=(--conda-prefix "${snakemake_conda_prefix}")
 fi
 
+test_workflow_jobs="${TEST_WORKFLOW_JOBS:-200}"
+
 env_snakemake_args=()
 if [[ -n "${TEST_WORKFLOW_SNAKEMAKE_ARGS:-}" ]]; then
     # Intended for simple Snakemake CLI tokens, such as --set-resources entries.
@@ -76,7 +84,7 @@ snakemake \
     "${conda_prefix_args[@]}" \
     --rerun-incomplete \
     --retries 2 \
-    --jobs 200 \
+    --jobs "${test_workflow_jobs}" \
     --latency-wait 600 \
     "${env_snakemake_args[@]}" \
     "${extra_snakemake_args[@]}"

From 6fd4ee2969015866890e5e722617f59628072050 Mon Sep 17 00:00:00 2001
From: Adam Freedman <adamfreedman@fas.harvard.edu>
Date: Wed, 10 Jun 2026 15:52:59 -0400
Subject: [PATCH 2/6] Use CellBender test shim in hosted full workflow action

---
 .github/workflows/full-workflow.yml |  4 +++
 tests/README.md                     |  2 ++
 tests/bin/cellbender                | 56 +++++++++++++++++++++++++++++
 3 files changed, 62 insertions(+)
 create mode 100755 tests/bin/cellbender

diff --git a/.github/workflows/full-workflow.yml b/.github/workflows/full-workflow.yml
index 34c8e95..1069ecf 100644
--- a/.github/workflows/full-workflow.yml
+++ b/.github/workflows/full-workflow.yml
@@ -27,6 +27,7 @@ jobs:
         --executor local
         --cores 2
       TEST_WORKFLOW_JOBS: "2"
+      TEST_WORKFLOW_FAKE_CELLBENDER_SOURCE: ${{ github.workspace }}/tests/reference_outputs/testdata/results/cellbender
       TEST_WORKFLOW_SNAKEMAKE_ARGS: >-
         --resources mem_mb=8192
         --set-resources
@@ -68,6 +69,9 @@ jobs:
       - name: Check out repository
         uses: actions/checkout@v4
 
+      - name: Add test command shims to PATH
+        run: echo "${GITHUB_WORKSPACE}/tests/bin" >> "${GITHUB_PATH}"
+
       - name: Create test environment
         uses: mamba-org/setup-micromamba@v2
         with:
diff --git a/tests/README.md b/tests/README.md
index bffdefe..2377941 100644
--- a/tests/README.md
+++ b/tests/README.md
@@ -52,6 +52,8 @@ The full-run test calls `tests/run_test_workflow.sh`, which uses `testdata/sampl
 
 For testing, omit `--snakemake-conda-prefix` so Snakemake uses its default `.snakemake/conda` location under the repository root. The runner assumes that the current environment already provides `snakemake` on `PATH`.
 
+The GitHub-hosted full workflow action uses `tests/bin/cellbender` as a test shim for the CellBender rule because hosted runners do not provide the GPU-enabled CellBender container runtime used on the cluster. The shim copies the reference CellBender H5 outputs into the expected rule outputs, while the rest of the workflow runs normally and is still compared against the reference snapshot. Cluster/HPC runs of `pytest tests --run-workflow` do not use this shim unless `tests/bin` is explicitly added to `PATH`.
+
 
 ## Reference outputs
 
diff --git a/tests/bin/cellbender b/tests/bin/cellbender
new file mode 100755
index 0000000..dc5f1da
--- /dev/null
+++ b/tests/bin/cellbender
@@ -0,0 +1,56 @@
+#!/usr/bin/env bash
+set -euo pipefail
+
+if [[ "${1:-}" != "remove-background" ]]; then
+    echo "test CellBender shim only supports: cellbender remove-background" >&2
+    exit 2
+fi
+shift
+
+if [[ "${1:-}" == "--help" ]]; then
+    echo "Usage: cellbender remove-background --input INPUT --output OUTPUT [--seed SEED] [--cuda]"
+    exit 0
+fi
+
+output=""
+while [[ $# -gt 0 ]]; do
+    case "$1" in
+        --output)
+            output="${2:-}"
+            shift 2
+            ;;
+        --input|--seed)
+            shift 2
+            ;;
+        --cuda)
+            shift
+            ;;
+        *)
+            shift
+            ;;
+    esac
+done
+
+if [[ -z "${output}" ]]; then
+    echo "test CellBender shim requires --output" >&2
+    exit 2
+fi
+
+source_dir="${TEST_WORKFLOW_FAKE_CELLBENDER_SOURCE:-}"
+if [[ -z "${source_dir}" ]]; then
+    echo "TEST_WORKFLOW_FAKE_CELLBENDER_SOURCE is required for the test CellBender shim" >&2
+    exit 127
+fi
+
+base_source="${source_dir}/cellbender_test.h5"
+filtered_source="${source_dir}/cellbender_test_filtered.h5"
+filtered_output="${output%.h5}_filtered.h5"
+
+if [[ ! -s "${base_source}" || ! -s "${filtered_source}" ]]; then
+    echo "test CellBender shim could not find reference H5 outputs under ${source_dir}" >&2
+    exit 1
+fi
+
+mkdir -p "$(dirname "${output}")"
+cp "${base_source}" "${output}"
+cp "${filtered_source}" "${filtered_output}"

From c9c097d077871c792d9ac927bb187a4312294acd Mon Sep 17 00:00:00 2001
From: Adam Freedman <adamfreedman@fas.harvard.edu>
Date: Wed, 10 Jun 2026 16:34:00 -0400
Subject: [PATCH 3/6] Enforce test workflow seed and scope metadata tolerance

---
 tests/README.md                 |  2 +-
 tests/compare_seurat_metadata.R | 25 ++++++++++++++++++++-----
 tests/run_test_workflow.sh      | 20 ++++++++++++++++++++
 3 files changed, 41 insertions(+), 6 deletions(-)

diff --git a/tests/README.md b/tests/README.md
index 2377941..c116bb4 100644
--- a/tests/README.md
+++ b/tests/README.md
@@ -57,7 +57,7 @@ The GitHub-hosted full workflow action uses `tests/bin/cellbender` as a test shi
 
 ## Reference outputs
 
-The full workflow test compares regenerated files in `testdata/results/` against reference files under `tests/reference_outputs/`. The compared file list is in `tests/test_reference_output_files.txt`. Seurat `.rds` files are compared at the metadata-table level, marker CSVs are compared by columns and `(cluster, genesymbol)` rows with numeric tolerance, emptyDrops matrix files are compared after gzip decompression, CellBender H5 outputs are compared with `h5diff`, and remaining durable outputs are compared byte-for-byte.
+The full workflow test compares regenerated files in `testdata/results/` against reference files under `tests/reference_outputs/`. The compared file list is in `tests/test_reference_output_files.txt`. Seurat `.rds` files are compared at the metadata-table level, marker CSVs are compared by columns and `(cluster, genesymbol)` rows with numeric tolerance, emptyDrops matrix files are compared after gzip decompression, CellBender H5 outputs are compared with `h5diff`, and remaining durable outputs are compared byte-for-byte. Seurat metadata numeric columns use a strict default tolerance, except `neighborhood_purity`, which allows a small absolute tolerance because it is a derived nearest-neighbor purity metric that can vary slightly across R/Bioconductor/platform builds even when barcode order and `seurat_clusters` match. Override that tolerance with `SEURAT_METADATA_NEIGHBORHOOD_PURITY_TOLERANCE` if needed.
 
 To refresh the reference snapshot after intentionally changing workflow behavior, first run the full test workflow so `testdata/results/` contains the desired outputs, then run:
 
diff --git a/tests/compare_seurat_metadata.R b/tests/compare_seurat_metadata.R
index 6a2e638..249c58b 100644
--- a/tests/compare_seurat_metadata.R
+++ b/tests/compare_seurat_metadata.R
@@ -6,10 +6,19 @@ if (length(args) == 0 || length(args) %% 2 != 0) {
 suppressPackageStartupMessages(library("Seurat"))
 
 numeric_tolerance <- as.numeric(Sys.getenv("SEURAT_METADATA_NUMERIC_TOLERANCE", "1e-8"))
-if (is.na(numeric_tolerance) || length(numeric_tolerance) != 1) {
-  stop("SEURAT_METADATA_NUMERIC_TOLERANCE must be numeric", call. = FALSE)
+if (is.na(numeric_tolerance) || length(numeric_tolerance) != 1 || numeric_tolerance < 0) {
+  stop("SEURAT_METADATA_NUMERIC_TOLERANCE must be a non-negative number", call. = FALSE)
 }
 
+neighborhood_purity_tolerance <- as.numeric(Sys.getenv("SEURAT_METADATA_NEIGHBORHOOD_PURITY_TOLERANCE", "0.02"))
+if (is.na(neighborhood_purity_tolerance) || length(neighborhood_purity_tolerance) != 1 || neighborhood_purity_tolerance < 0) {
+  stop("SEURAT_METADATA_NEIGHBORHOOD_PURITY_TOLERANCE must be a non-negative number", call. = FALSE)
+}
+
+metadata_column_tolerances <- c(
+  neighborhood_purity = neighborhood_purity_tolerance
+)
+
 fail <- function(path, message) {
   stop(sprintf("%s: %s", path, message), call. = FALSE)
 }
@@ -30,14 +39,20 @@ compare_metadata_column <- function(current_path, column, current_values, refere
     if (any(comparable)) {
       diff <- abs(current_numeric[comparable] - reference_numeric[comparable])
       scale <- pmax(abs(current_numeric[comparable]), abs(reference_numeric[comparable]), 1)
-      bad <- diff > numeric_tolerance * scale
+      column_tolerance <- if (column %in% names(metadata_column_tolerances)) {
+        metadata_column_tolerances[[column]]
+      } else {
+        numeric_tolerance
+      }
+      bad <- diff > column_tolerance * scale
       if (any(bad)) {
         fail(
           current_path,
           sprintf(
-            "metadata column %s differs numerically; max abs diff %.12g",
+            "metadata column %s differs numerically; max abs diff %.12g exceeds tolerance %.12g",
             column,
-            max(diff)
+            max(diff),
+            column_tolerance
           )
         )
       }
diff --git a/tests/run_test_workflow.sh b/tests/run_test_workflow.sh
index 07207eb..60fcc22 100755
--- a/tests/run_test_workflow.sh
+++ b/tests/run_test_workflow.sh
@@ -49,6 +49,26 @@ if ! command -v snakemake >/dev/null 2>&1; then
     exit 127
 fi
 
+workflow_seed=$(python - <<PYSEED
+from pathlib import Path
+import re
+
+config_text = Path("config/config.yaml").read_text()
+for line in config_text.splitlines():
+    match = re.match(r"^workflow_seed\s*:\s*([^#\s]+)", line)
+    if match:
+        print(int(match.group(1)))
+        break
+else:
+    raise SystemExit("workflow_seed is missing from config/config.yaml")
+PYSEED
+)
+export SCRNASEQ_PREPROCESS_SEED="${SCRNASEQ_PREPROCESS_SEED:-${workflow_seed}}"
+if [[ "${SCRNASEQ_PREPROCESS_SEED}" != "${workflow_seed}" ]]; then
+    echo "SCRNASEQ_PREPROCESS_SEED=${SCRNASEQ_PREPROCESS_SEED} does not match config workflow_seed=${workflow_seed}" >&2
+    exit 2
+fi
+
 profile_args=(--workflow-profile profiles/slurm --profile cannon)
 if [[ -n "${TEST_WORKFLOW_PROFILE_ARGS:-}" ]]; then
     # Intended for simple Snakemake profile/executor args.

From 0ccb1e95dee489f89b29376e11bf0f46ff468510 Mon Sep 17 00:00:00 2001
From: Adam Freedman <adamfreedman@fas.harvard.edu>
Date: Thu, 11 Jun 2026 06:37:01 -0400
Subject: [PATCH 4/6] Pin workflow conda envs and CellBender container

---
 .github/workflows/full-workflow.yml           |   2 +-
 .github/workflows/tests.yml                   |   3 +-
 tests/README.md                               |   6 +-
 tests/test_conda_container_validation.py      |  40 +-
 workflow/envs/cellbender.yml                  |  15 -
 workflow/envs/doubletfinder.linux-64.pin.txt  | 337 +++++++++++++++++
 workflow/envs/emptydrops.linux-64.pin.txt     | 342 +++++++++++++++++
 workflow/envs/posthocfilter.linux-64.pin.txt  | 334 ++++++++++++++++
 workflow/envs/scdblfinder.linux-64.pin.txt    | 357 +++++++++++++++++
 workflow/envs/soupx.linux-64.pin.txt          | 322 ++++++++++++++++
 workflow/envs/tenx2seuratrds.linux-64.pin.txt | 358 ++++++++++++++++++
 workflow/rules/cellbender.smk                 |   2 +-
 12 files changed, 2088 insertions(+), 30 deletions(-)
 delete mode 100644 workflow/envs/cellbender.yml
 create mode 100644 workflow/envs/doubletfinder.linux-64.pin.txt
 create mode 100644 workflow/envs/emptydrops.linux-64.pin.txt
 create mode 100644 workflow/envs/posthocfilter.linux-64.pin.txt
 create mode 100644 workflow/envs/scdblfinder.linux-64.pin.txt
 create mode 100644 workflow/envs/soupx.linux-64.pin.txt
 create mode 100644 workflow/envs/tenx2seuratrds.linux-64.pin.txt

diff --git a/.github/workflows/full-workflow.yml b/.github/workflows/full-workflow.yml
index 1069ecf..daefa86 100644
--- a/.github/workflows/full-workflow.yml
+++ b/.github/workflows/full-workflow.yml
@@ -83,7 +83,7 @@ jobs:
         uses: actions/cache@v4
         with:
           path: .snakemake/conda
-          key: snakemake-conda-full-${{ runner.os }}-${{ hashFiles('workflow/envs/*.yml') }}
+          key: snakemake-conda-full-${{ runner.os }}-${{ hashFiles('workflow/envs/*.yml', 'workflow/envs/*.pin.txt') }}
           restore-keys: |
             snakemake-conda-full-${{ runner.os }}-
             snakemake-conda-${{ runner.os }}-
diff --git a/.github/workflows/tests.yml b/.github/workflows/tests.yml
index 2cfa65d..cb2eb9d 100644
--- a/.github/workflows/tests.yml
+++ b/.github/workflows/tests.yml
@@ -44,7 +44,7 @@ jobs:
         uses: actions/cache@v4
         with:
           path: .snakemake/conda
-          key: snakemake-conda-${{ runner.os }}-${{ hashFiles('workflow/envs/*.yml') }}
+          key: snakemake-conda-${{ runner.os }}-${{ hashFiles('workflow/envs/*.yml', 'workflow/envs/*.pin.txt') }}
           restore-keys: |
             snakemake-conda-${{ runner.os }}-
 
@@ -58,7 +58,6 @@ jobs:
       fail-fast: false
       matrix:
         env_name:
-          - cellbender.yml
           - doubletfinder.yml
           - emptydrops.yml
           - posthocfilter.yml
diff --git a/tests/README.md b/tests/README.md
index c116bb4..691bb75 100644
--- a/tests/README.md
+++ b/tests/README.md
@@ -24,7 +24,7 @@ GitHub Actions runs the default test suite via `.github/workflows/tests.yml`, us
 
 The default test suite also includes a focused local rule-execution smoke test. It runs the real `tenx2seuratrds`, `find_markers`, and `combine_markers` rule chain against `testdata/`, using Snakemake's `--use-conda` support and writing outputs under pytest's temporary directory. This catches broken R package imports, script argument drift, invalid Seurat object creation, and marker CSV schema changes without submitting to SLURM.
 
-The R output validator also checks that the Seurat object has at least 100 features and 100 cells; metadata rows match the cell count; barcode row names are present, unique, and nonempty; `orig.ident`, `nCount_RNA`, `nFeature_RNA`, `percent.mt`, and `seurat_clusters` metadata columns exist; RNA count and feature-count metadata values are finite and positive; mitochondrial percentages are finite and within `[0, 100]`; at least two clusters are present; PCA and UMAP reductions exist; the marker table is nonempty and has the expected columns; marker gene symbols are present and nonempty; marker numeric columns are finite; marker p-value and percent columns are within `[0, 1]`; marker clusters are present in the Seurat metadata; markers are reported for at least two clusters; and the marker `workflow` column matches the expected test workflow label. The test runner environment is defined in the repository-level `environment.yml`; the rule-specific R environment is still created by Snakemake from `workflow/envs/tenx2seuratrds.yml`. A separate lightweight checkpoint-expansion test uses a fake `Rscript` to materialize the `marker_manifest` checkpoint, verify that dynamic `find_markers` jobs are generated for each cluster id, and confirm that `combine_markers` receives the expected marker chunks.
+The R output validator also checks that the Seurat object has at least 100 features and 100 cells; metadata rows match the cell count; barcode row names are present, unique, and nonempty; `orig.ident`, `nCount_RNA`, `nFeature_RNA`, `percent.mt`, and `seurat_clusters` metadata columns exist; RNA count and feature-count metadata values are finite and positive; mitochondrial percentages are finite and within `[0, 100]`; at least two clusters are present; PCA and UMAP reductions exist; the marker table is nonempty and has the expected columns; marker gene symbols are present and nonempty; marker numeric columns are finite; marker p-value and percent columns are within `[0, 1]`; marker clusters are present in the Seurat metadata; markers are reported for at least two clusters; and the marker `workflow` column matches the expected test workflow label. The test runner environment is defined in the repository-level `environment.yml`; the rule-specific R environment is still created by Snakemake from `workflow/envs/tenx2seuratrds.yml`, using `workflow/envs/tenx2seuratrds.linux-64.pin.txt` on Linux to recreate the locked package set. A separate lightweight checkpoint-expansion test uses a fake `Rscript` to materialize the `marker_manifest` checkpoint, verify that dynamic `find_markers` jobs are generated for each cluster id, and confirm that `combine_markers` receives the expected marker chunks.
 
 
 ## Optional conda and container validation
@@ -37,7 +37,7 @@ pytest tests --run-container-validation
 pytest tests --run-doubletfinder-install
 ```
 
-`--run-conda-validation` creates each `workflow/envs/*.yml` environment in a pytest temporary directory and checks key R/Python package imports. To validate only one environment, pass `--conda-env-name ENV_FILE`, for example `--conda-env-name soupx.yml`; GitHub Actions uses this selector to run conda validation as one matrix job per env. `--run-container-validation` pulls the CellBender container with Docker, Apptainer, or Singularity. `--run-doubletfinder-install` runs the networked Snakemake `install_doubletfinder` rule and confirms that `DoubletFinder` can be imported from the created rule environment.
+`--run-conda-validation` creates each `workflow/envs/*.yml` environment in a pytest temporary directory and checks key R/Python package imports; on Linux it creates environments from the matching `*.linux-64.pin.txt` files so validation exercises the locked package specs used by Snakemake. To validate only one environment, pass `--conda-env-name ENV_FILE`, for example `--conda-env-name soupx.yml`; GitHub Actions uses this selector to run conda validation as one matrix job per env. `--run-container-validation` pulls the pinned CellBender container digest with Docker, Apptainer, or Singularity. `--run-doubletfinder-install` runs the networked Snakemake `install_doubletfinder` rule and confirms that `DoubletFinder` can be imported from the created rule environment.
 
 
 ## Optional full workflow run
@@ -50,7 +50,7 @@ pytest tests --run-workflow
 
 The full-run test calls `tests/run_test_workflow.sh`, which uses `testdata/samplesheet_test.tsv` and overrides the workflow output directory with `resultsDir=testdata/results`. The manifest in `tests/test_sample_rule_output_files.txt` is therefore written with paths under `testdata/results/`.
 
-For testing, omit `--snakemake-conda-prefix` so Snakemake uses its default `.snakemake/conda` location under the repository root. The runner assumes that the current environment already provides `snakemake` on `PATH`.
+For testing, omit `--snakemake-conda-prefix` so Snakemake uses its default `.snakemake/conda` location under the repository root. Snakemake includes the `*.linux-64.pin.txt` contents in environment hashes, so changing a lock file creates a new rule environment. The runner assumes that the current environment already provides `snakemake` on `PATH`.
 
 The GitHub-hosted full workflow action uses `tests/bin/cellbender` as a test shim for the CellBender rule because hosted runners do not provide the GPU-enabled CellBender container runtime used on the cluster. The shim copies the reference CellBender H5 outputs into the expected rule outputs, while the rest of the workflow runs normally and is still compared against the reference snapshot. Cluster/HPC runs of `pytest tests --run-workflow` do not use this shim unless `tests/bin` is explicitly added to `PATH`.
 
diff --git a/tests/test_conda_container_validation.py b/tests/test_conda_container_validation.py
index f3893bb..09dda54 100644
--- a/tests/test_conda_container_validation.py
+++ b/tests/test_conda_container_validation.py
@@ -1,4 +1,5 @@
 import os
+import platform
 import re
 import shutil
 import subprocess
@@ -12,7 +13,6 @@
 RULE_DIR = Path("workflow/rules")
 TEST_SAMPLE_SHEET = Path("testdata/samplesheet_test.tsv")
 EXPECTED_ENV_FILES = {
-    "cellbender.yml",
     "doubletfinder.yml",
     "emptydrops.yml",
     "posthocfilter.yml",
@@ -22,9 +22,9 @@
 }
 CONDA_REFERENCE_RE = re.compile(r"conda:\s*\n\s*['\"]\.\./envs/([^'\"]+)['\"]")
 CONTAINER_RE = re.compile(r"container:\s*\n\s*['\"]([^'\"]+)['\"]")
+CELLBENDER_CONTAINER_URI = "docker://us.gcr.io/broad-dsde-methods/cellbender@sha256:093f2caf1ce4acae4541ea45e52ab7b220ca131ec73b4d1f664b85fe12850bae"
 
 R_IMPORTS_BY_ENV = {
-    "cellbender.yml": ["Seurat", "tidyverse", "bluster"],
     "doubletfinder.yml": ["Seurat", "tidyverse", "remotes", "fields", "Matrix", "KernSmooth", "ROCR", "igraph", "glmGamPoi", "bluster"],
     "emptydrops.yml": ["Seurat", "tidyverse", "DropletUtils", "scater", "glmGamPoi", "bluster", "Matrix", "R.utils"],
     "posthocfilter.yml": ["Seurat", "tidyverse", "glmGamPoi", "scater", "bluster"],
@@ -32,9 +32,7 @@
     "soupx.yml": ["Seurat", "tidyverse", "glmGamPoi", "bluster", "SoupX"],
     "tenx2seuratrds.yml": ["Seurat", "tidyverse", "scCustomize", "hdf5r", "glmGamPoi", "bluster", "presto"],
 }
-PYTHON_IMPORTS_BY_ENV = {
-    "cellbender.yml": ["cellbender"],
-}
+PYTHON_IMPORTS_BY_ENV = {}
 
 
 def repo_root():
@@ -73,6 +71,19 @@ def available_conda_frontend():
     raise AssertionError("neither mamba nor conda is available on PATH")
 
 
+def linux_conda_subdir():
+    if platform.system() == "Linux" and platform.machine() in {"x86_64", "AMD64"}:
+        return "linux-64"
+    return None
+
+
+def pin_file_for_env(env_path):
+    subdir = linux_conda_subdir()
+    if subdir is None:
+        return None
+    return env_path.with_suffix(f".{subdir}.pin.txt")
+
+
 def r_require_namespace_expr(packages):
     package_vector = ", ".join(repr(package) for package in packages)
     return (
@@ -100,6 +111,13 @@ def test_workflow_conda_env_files_are_well_formed_and_referenced_envs_exist():
         assert env.get("channel_priority") == "strict", env_path
         assert env.get("dependencies"), env_path
 
+        pin_path = pin_file_for_env(env_path)
+        if pin_path is not None:
+            assert pin_path.exists(), f"missing Snakemake conda pin file: {pin_path}"
+            pin_text = pin_path.read_text()
+            assert "@EXPLICIT" in pin_text, f"pin file is not an explicit conda spec: {pin_path}"
+            assert "conda-forge" in pin_text or "bioconda" in pin_text, pin_path
+
     referenced = set()
     for rule_path in sorted((root / RULE_DIR).glob("*.smk")):
         referenced.update(CONDA_REFERENCE_RE.findall(rule_path.read_text()))
@@ -116,7 +134,7 @@ def test_workflow_container_declarations_are_explicit_and_recognized():
             containers.append((rule_path, uri))
 
     assert containers, "no workflow container declarations found"
-    assert containers == [(root / "workflow/rules/cellbender.smk", "docker://us.gcr.io/broad-dsde-methods/cellbender:latest")]
+    assert containers == [(root / "workflow/rules/cellbender.smk", CELLBENDER_CONTAINER_URI)]
     for _, uri in containers:
         assert uri.startswith("docker://")
         assert ":" in uri.removeprefix("docker://"), f"container URI is missing a tag: {uri}"
@@ -135,8 +153,14 @@ def test_conda_env_solves_and_key_packages_import(tmp_path, pytestconfig, env_na
     prefix = tmp_path / env_name.removesuffix(".yml")
     conda = available_conda_frontend()
 
+    pin_path = pin_file_for_env(env_path)
+    if pin_path is not None and pin_path.exists():
+        create_cmd = [conda, "create", "--yes", "--prefix", str(prefix), "--file", str(pin_path)]
+    else:
+        create_cmd = [conda, "env", "create", "--yes", "--prefix", str(prefix), "--file", str(env_path)]
+
     create = run_command(
-        [conda, "env", "create", "--yes", "--prefix", str(prefix), "--file", str(env_path)],
+        create_cmd,
         root,
         timeout=1800,
     )
@@ -172,7 +196,7 @@ def test_cellbender_container_can_be_pulled(tmp_path, pytestconfig):
         pytest.skip("use --run-container-validation to pull workflow containers")
 
     root = repo_root()
-    uri = "docker://us.gcr.io/broad-dsde-methods/cellbender:latest"
+    uri = CELLBENDER_CONTAINER_URI
     docker_image = uri.removeprefix("docker://")
 
     if shutil.which("docker"):
diff --git a/workflow/envs/cellbender.yml b/workflow/envs/cellbender.yml
deleted file mode 100644
index da1aef2..0000000
--- a/workflow/envs/cellbender.yml
+++ /dev/null
@@ -1,15 +0,0 @@
-channels:
- - conda-forge
- - bioconda
-
-channel_priority: strict
-
-dependencies:
-  - r-tidyverse
-  - python=3.9  
-  - r-base
-  - r-seurat
-  - bioconductor-bluster
-  - pip          
-  - pip:       
-      - cellbender
diff --git a/workflow/envs/doubletfinder.linux-64.pin.txt b/workflow/envs/doubletfinder.linux-64.pin.txt
new file mode 100644
index 0000000..308bb41
--- /dev/null
+++ b/workflow/envs/doubletfinder.linux-64.pin.txt
@@ -0,0 +1,337 @@
+# This file may be used to create an environment using:
+# $ conda create --name <env> --file <this file>
+# platform: linux-64
+# created-by: conda 25.11.0
+@EXPLICIT
+https://conda.anaconda.org/conda-forge/noarch/_r-mutex-1.0.1-anacondar_1.tar.bz2
+https://conda.anaconda.org/conda-forge/noarch/ca-certificates-2026.5.20-hbd8a1cb_0.conda
+https://conda.anaconda.org/conda-forge/noarch/font-ttf-dejavu-sans-mono-2.37-hab24e00_0.tar.bz2
+https://conda.anaconda.org/conda-forge/noarch/font-ttf-inconsolata-3.000-h77eed37_0.tar.bz2
+https://conda.anaconda.org/conda-forge/noarch/font-ttf-source-code-pro-2.038-h77eed37_0.tar.bz2
+https://conda.anaconda.org/conda-forge/noarch/font-ttf-ubuntu-0.83-h77eed37_3.conda
+https://conda.anaconda.org/conda-forge/noarch/kernel-headers_linux-64-4.18.0-he073ed8_9.conda
+https://conda.anaconda.org/conda-forge/noarch/libgcc-devel_linux-64-15.2.0-hcc6f6b0_119.conda
+https://conda.anaconda.org/conda-forge/linux-64/libgomp-15.2.0-he0feb66_19.conda
+https://conda.anaconda.org/conda-forge/noarch/libstdcxx-devel_linux-64-15.2.0-hd446a21_119.conda
+https://conda.anaconda.org/conda-forge/linux-64/libzlib-1.3.2-h25fd6f3_2.conda
+https://conda.anaconda.org/conda-forge/linux-64/pandoc-3.9.0.2-ha770c72_0.conda
+https://conda.anaconda.org/conda-forge/noarch/tzdata-2025c-hc9c84f9_1.conda
+https://conda.anaconda.org/conda-forge/linux-64/_openmp_mutex-4.5-20_gnu.conda
+https://conda.anaconda.org/conda-forge/noarch/fonts-conda-forge-1-hc364b38_1.conda
+https://conda.anaconda.org/conda-forge/noarch/sysroot_linux-64-2.28-h4ee821c_9.conda
+https://conda.anaconda.org/conda-forge/linux-64/zlib-1.3.2-h25fd6f3_2.conda
+https://conda.anaconda.org/conda-forge/linux-64/zstd-1.5.7-hb78ec9c_6.conda
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diff --git a/workflow/envs/emptydrops.linux-64.pin.txt b/workflow/envs/emptydrops.linux-64.pin.txt
new file mode 100644
index 0000000..0eeef35
--- /dev/null
+++ b/workflow/envs/emptydrops.linux-64.pin.txt
@@ -0,0 +1,342 @@
+# This file may be used to create an environment using:
+# $ conda create --name <env> --file <this file>
+# platform: linux-64
+# created-by: conda 25.11.0
+@EXPLICIT
+https://conda.anaconda.org/conda-forge/noarch/_r-mutex-1.0.1-anacondar_1.tar.bz2
+https://conda.anaconda.org/conda-forge/noarch/ca-certificates-2026.5.20-hbd8a1cb_0.conda
+https://conda.anaconda.org/conda-forge/noarch/font-ttf-dejavu-sans-mono-2.37-hab24e00_0.tar.bz2
+https://conda.anaconda.org/conda-forge/noarch/font-ttf-inconsolata-3.000-h77eed37_0.tar.bz2
+https://conda.anaconda.org/conda-forge/noarch/font-ttf-source-code-pro-2.038-h77eed37_0.tar.bz2
+https://conda.anaconda.org/conda-forge/noarch/font-ttf-ubuntu-0.83-h77eed37_3.conda
+https://conda.anaconda.org/conda-forge/noarch/kernel-headers_linux-64-4.18.0-he073ed8_9.conda
+https://conda.anaconda.org/conda-forge/noarch/libgcc-devel_linux-64-15.2.0-hcc6f6b0_119.conda
+https://conda.anaconda.org/conda-forge/linux-64/libgomp-15.2.0-he0feb66_19.conda
+https://conda.anaconda.org/conda-forge/noarch/libstdcxx-devel_linux-64-15.2.0-hd446a21_119.conda
+https://conda.anaconda.org/conda-forge/linux-64/libzlib-1.3.2-h25fd6f3_2.conda
+https://conda.anaconda.org/conda-forge/linux-64/pandoc-3.9.0.2-ha770c72_0.conda
+https://conda.anaconda.org/conda-forge/noarch/tzdata-2025c-hc9c84f9_1.conda
+https://conda.anaconda.org/conda-forge/linux-64/_openmp_mutex-4.5-20_gnu.conda
+https://conda.anaconda.org/conda-forge/noarch/fonts-conda-forge-1-hc364b38_1.conda
+https://conda.anaconda.org/conda-forge/noarch/sysroot_linux-64-2.28-h4ee821c_9.conda
+https://conda.anaconda.org/conda-forge/linux-64/zstd-1.5.7-hb78ec9c_6.conda
+https://conda.anaconda.org/conda-forge/noarch/fonts-conda-ecosystem-1-0.tar.bz2
+https://conda.anaconda.org/conda-forge/linux-64/ld_impl_linux-64-2.45.1-default_hbd61a6d_102.conda
+https://conda.anaconda.org/conda-forge/linux-64/libgcc-15.2.0-he0feb66_19.conda
+https://conda.anaconda.org/conda-forge/linux-64/binutils_impl_linux-64-2.45.1-default_hfdba357_102.conda
+https://conda.anaconda.org/conda-forge/linux-64/bzip2-1.0.8-hda65f42_9.conda
+https://conda.anaconda.org/conda-forge/linux-64/c-ares-1.34.6-hb03c661_0.conda
+https://conda.anaconda.org/conda-forge/linux-64/fribidi-1.0.16-hb03c661_0.conda
+https://conda.anaconda.org/conda-forge/linux-64/keyutils-1.6.3-hb9d3cd8_0.conda
+https://conda.anaconda.org/conda-forge/linux-64/libdeflate-1.25-h17f619e_0.conda
+https://conda.anaconda.org/conda-forge/linux-64/libexpat-2.8.1-hecca717_0.conda
+https://conda.anaconda.org/conda-forge/linux-64/libffi-3.5.2-h3435931_0.conda
+https://conda.anaconda.org/conda-forge/linux-64/libgcc-ng-15.2.0-h69a702a_19.conda
+https://conda.anaconda.org/conda-forge/linux-64/libgfortran5-15.2.0-h68bc16d_19.conda
+https://conda.anaconda.org/conda-forge/linux-64/libiconv-1.18-h3b78370_2.conda
+https://conda.anaconda.org/conda-forge/linux-64/libjpeg-turbo-3.1.4.1-hb03c661_0.conda
+https://conda.anaconda.org/conda-forge/linux-64/liblzma-5.8.3-hb03c661_0.conda
+https://conda.anaconda.org/conda-forge/linux-64/libpng-1.6.58-h421ea60_0.conda
+https://conda.anaconda.org/conda-forge/linux-64/libstdcxx-15.2.0-h934c35e_19.conda
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diff --git a/workflow/envs/posthocfilter.linux-64.pin.txt b/workflow/envs/posthocfilter.linux-64.pin.txt
new file mode 100644
index 0000000..34c517a
--- /dev/null
+++ b/workflow/envs/posthocfilter.linux-64.pin.txt
@@ -0,0 +1,334 @@
+# This file may be used to create an environment using:
+# $ conda create --name <env> --file <this file>
+# platform: linux-64
+# created-by: conda 25.11.0
+@EXPLICIT
+https://conda.anaconda.org/conda-forge/noarch/_r-mutex-1.0.1-anacondar_1.tar.bz2
+https://conda.anaconda.org/conda-forge/noarch/ca-certificates-2026.5.20-hbd8a1cb_0.conda
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diff --git a/workflow/envs/scdblfinder.linux-64.pin.txt b/workflow/envs/scdblfinder.linux-64.pin.txt
new file mode 100644
index 0000000..015efe4
--- /dev/null
+++ b/workflow/envs/scdblfinder.linux-64.pin.txt
@@ -0,0 +1,357 @@
+# This file may be used to create an environment using:
+# $ conda create --name <env> --file <this file>
+# platform: linux-64
+# created-by: conda 25.11.0
+@EXPLICIT
+https://conda.anaconda.org/conda-forge/noarch/_r-mutex-1.0.1-anacondar_1.tar.bz2
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diff --git a/workflow/envs/soupx.linux-64.pin.txt b/workflow/envs/soupx.linux-64.pin.txt
new file mode 100644
index 0000000..c76e97b
--- /dev/null
+++ b/workflow/envs/soupx.linux-64.pin.txt
@@ -0,0 +1,322 @@
+# This file may be used to create an environment using:
+# $ conda create --name <env> --file <this file>
+# platform: linux-64
+# created-by: conda 25.11.0
+@EXPLICIT
+https://conda.anaconda.org/conda-forge/noarch/_r-mutex-1.0.1-anacondar_1.tar.bz2
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diff --git a/workflow/envs/tenx2seuratrds.linux-64.pin.txt b/workflow/envs/tenx2seuratrds.linux-64.pin.txt
new file mode 100644
index 0000000..c7f1643
--- /dev/null
+++ b/workflow/envs/tenx2seuratrds.linux-64.pin.txt
@@ -0,0 +1,358 @@
+# This file may be used to create an environment using:
+# $ conda create --name <env> --file <this file>
+# platform: linux-64
+# created-by: conda 25.11.0
+@EXPLICIT
+https://conda.anaconda.org/conda-forge/noarch/_r-mutex-1.0.1-anacondar_1.tar.bz2
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diff --git a/workflow/rules/cellbender.smk b/workflow/rules/cellbender.smk
index 5b9d8de..ca848d2 100644
--- a/workflow/rules/cellbender.smk
+++ b/workflow/rules/cellbender.smk
@@ -11,7 +11,7 @@ rule cellbender:
         workdir="scratch/cellbender/{sample}",
         seed=WORKFLOW_SEED
     container:
-        "docker://us.gcr.io/broad-dsde-methods/cellbender:latest"
+        "docker://us.gcr.io/broad-dsde-methods/cellbender@sha256:093f2caf1ce4acae4541ea45e52ab7b220ca131ec73b4d1f664b85fe12850bae"
     resources:
         mem_mb = lambda wildcards, attempt: int(50000 * (2 ** (attempt - 1))),
         slurm_partition = "gpu",

From 6469ca94b3541cf79c08c61568d7ddf8cad76ad3 Mon Sep 17 00:00:00 2001
From: Adam Freedman <adamfreedman@fas.harvard.edu>
Date: Thu, 11 Jun 2026 15:58:42 -0400
Subject: [PATCH 5/6] Refresh pinned workflow reference outputs

---
 .../results/cellbender/cellbender_test.h5     | Bin 395027 -> 395027 bytes
 .../cellbender/cellbender_test_filtered.h5    | Bin 269840 -> 269840 bytes
 .../seurat_cellbender_fromraw_test.rds        | Bin 2617090 -> 2617090 bytes
 ..._doubletfinder_cellbender_fromraw_test.rds | Bin 2297178 -> 2297177 bytes
 ...at_doubletfinder_soupx_emptydrops_test.rds | Bin 2278942 -> 2278941 bytes
 .../seurat_doubletfinder_soupx_tenx_test.rds  | Bin 2602745 -> 2602745 bytes
 .../filtered_seurat_emptydrops_test.rds       | Bin 2783298 -> 2783298 bytes
 ..._doubletfinder_cellbender_fromraw_test.rds | Bin 2297184 -> 2297182 bytes
 ...ad_doubletfinder_soupx_emptydrops_test.rds | Bin 2278946 -> 2278946 bytes
 ...filt_mad_doubletfinder_soupx_tenx_test.rds | Bin 2602750 -> 2602750 bytes
 ...ad_scdblfinder_cellbender_fromraw_test.rds | Bin 2543988 -> 2543985 bytes
 ..._mad_scdblfinder_soupx_emptydrops_test.rds | Bin 2563379 -> 2563379 bytes
 ...ocfilt_mad_scdblfinder_soupx_tenx_test.rds | Bin 2721162 -> 2721164 bytes
 ..._doubletfinder_cellbender_fromraw_test.rds | Bin 1418065 -> 1418062 bytes
 ...ld_doubletfinder_soupx_emptydrops_test.rds | Bin 1492849 -> 1492850 bytes
 ...hreshold_doubletfinder_soupx_tenx_test.rds | Bin 1512039 -> 1512040 bytes
 ...ld_scdblfinder_cellbender_fromraw_test.rds | Bin 1639108 -> 1639109 bytes
 ...hold_scdblfinder_soupx_emptydrops_test.rds | Bin 1697522 -> 1697522 bytes
 ..._threshold_scdblfinder_soupx_tenx_test.rds | Bin 1556052 -> 1556051 bytes
 ...at_scdblfinder_cellbender_fromraw_test.rds | Bin 2543981 -> 2543980 bytes
 ...urat_scdblfinder_soupx_emptydrops_test.rds | Bin 2563374 -> 2563376 bytes
 .../seurat_scdblfinder_soupx_tenx_test.rds    | Bin 2721157 -> 2721158 bytes
 .../filtered_seurat_tenx_test.rds             | Bin 2860599 -> 2860604 bytes
 .../soupx/seurat_soupx_emptydrops_test.rds    | Bin 2586883 -> 2586883 bytes
 .../results/soupx/seurat_soupx_tenx_test.rds  | Bin 2839261 -> 2839260 bytes
 workflow/rules/install_doubletfinder.smk      |   2 +-
 26 files changed, 1 insertion(+), 1 deletion(-)

diff --git a/tests/reference_outputs/testdata/results/cellbender/cellbender_test.h5 b/tests/reference_outputs/testdata/results/cellbender/cellbender_test.h5
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diff --git a/tests/reference_outputs/testdata/results/cellbender/cellbender_test_filtered.h5 b/tests/reference_outputs/testdata/results/cellbender/cellbender_test_filtered.h5
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delta 1537
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diff --git a/tests/reference_outputs/testdata/results/doubletfinder/seurat_doubletfinder_cellbender_fromraw_test.rds b/tests/reference_outputs/testdata/results/doubletfinder/seurat_doubletfinder_cellbender_fromraw_test.rds
index 58f92edf8ff4ab7ceeac5a5a7ae01056c8535e32..69279bdd426e1530f48a685e979387e07c2066ed 100644
GIT binary patch
delta 22594
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diff --git a/tests/reference_outputs/testdata/results/doubletfinder/seurat_doubletfinder_soupx_emptydrops_test.rds b/tests/reference_outputs/testdata/results/doubletfinder/seurat_doubletfinder_soupx_emptydrops_test.rds
index 9ae38ddb9571631a80a4acb3bfe8b3f3932fc91f..43fab3e7cb8f32b63febf0c9a162a284f39a59d5 100644
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diff --git a/tests/reference_outputs/testdata/results/doubletfinder/seurat_doubletfinder_soupx_tenx_test.rds b/tests/reference_outputs/testdata/results/doubletfinder/seurat_doubletfinder_soupx_tenx_test.rds
index 3c544de0a60f9d1ea1d7d2fe39e40213ec293995..9a23888fdc7af598ab4683a1e6e9fd3c605539db 100644
GIT binary patch
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diff --git a/tests/reference_outputs/testdata/results/emptydrops/filtered_seurat_emptydrops_test.rds b/tests/reference_outputs/testdata/results/emptydrops/filtered_seurat_emptydrops_test.rds
index 8ab1f561e29cbfeb29aa8d4aa79f44c6a21265cc..99c3a33842d9b877cc9434dfb86e8eac87504258 100644
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diff --git a/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_mad_doubletfinder_cellbender_fromraw_test.rds b/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_mad_doubletfinder_cellbender_fromraw_test.rds
index 3efd8829ed01e20407ddf53906cdfabc5a70e1e2..b2d49ce50cc9dafba248a20305dcfe276e5b07b1 100644
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diff --git a/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_mad_doubletfinder_soupx_emptydrops_test.rds b/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_mad_doubletfinder_soupx_emptydrops_test.rds
index ddb1bf39b912fa121c7028628eb898da25ed1a80..c115c1356c78612dd66ff8e0d45b2546b810e181 100644
GIT binary patch
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diff --git a/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_mad_scdblfinder_soupx_emptydrops_test.rds b/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_mad_scdblfinder_soupx_emptydrops_test.rds
index 7dc21f83706acb5e9f8317103acfd920e2297e1e..cc10a7ca343c55825cfb9f86ba05c8645532e0dc 100644
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diff --git a/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_mad_scdblfinder_soupx_tenx_test.rds b/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_mad_scdblfinder_soupx_tenx_test.rds
index 7e2992462383a4bc13a8e59417c83cf48dbc24c9..4332e3b42accba3561468a09a33bd369c7556eb1 100644
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diff --git a/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_doubletfinder_cellbender_fromraw_test.rds b/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_doubletfinder_cellbender_fromraw_test.rds
index 897ac4463d615e4a3fd3cfdb34a70387b9781a9b..0b44341821400464bb17dd6385d252bb249ac21f 100644
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diff --git a/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_doubletfinder_soupx_emptydrops_test.rds b/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_doubletfinder_soupx_emptydrops_test.rds
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diff --git a/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_doubletfinder_soupx_tenx_test.rds b/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_doubletfinder_soupx_tenx_test.rds
index 5c888da99f18109abcae88a86d0deaa233a0b369..34111016853f0a3213d0323acb696e066a10dba1 100644
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diff --git a/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_scdblfinder_cellbender_fromraw_test.rds b/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_scdblfinder_cellbender_fromraw_test.rds
index aa3a9a40437c45e4b9f3a26bb78a7dab06ee03d1..67d768cc11f5212aeab6a9514f210c59b00a9b66 100644
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diff --git a/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_scdblfinder_soupx_emptydrops_test.rds b/tests/reference_outputs/testdata/results/posthocfilter/seurat_posthocfilt_threshold_scdblfinder_soupx_emptydrops_test.rds
index 5d65ea07ee66511776077db2b1b10f7a8b1cee0a..b7a3d42382f5777e32112f0238a9d53e68e89c83 100644
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index 5bb0c7490e156d6eae6499073d1cdde75fd646d8..8bfb5a19d31d9c0add7167885c8543407ea7a706 100644
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diff --git a/tests/reference_outputs/testdata/results/scdblfinder/seurat_scdblfinder_cellbender_fromraw_test.rds b/tests/reference_outputs/testdata/results/scdblfinder/seurat_scdblfinder_cellbender_fromraw_test.rds
index 93113aa3e1e41ea008f9d011f9ee8f37c21914f5..e43465d02475337ea1b75be88deab1bac883b222 100644
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diff --git a/tests/reference_outputs/testdata/results/scdblfinder/seurat_scdblfinder_soupx_emptydrops_test.rds b/tests/reference_outputs/testdata/results/scdblfinder/seurat_scdblfinder_soupx_emptydrops_test.rds
index a12609f8fa9eae3ec2894b0665f9dabab061c8dd..2d61abc599d3f7ba3a76aa66c978d1c26044b007 100644
GIT binary patch
delta 1118
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diff --git a/tests/reference_outputs/testdata/results/scdblfinder/seurat_scdblfinder_soupx_tenx_test.rds b/tests/reference_outputs/testdata/results/scdblfinder/seurat_scdblfinder_soupx_tenx_test.rds
index 56d02c4f32caa6e7ebca8cf2883cebd372bcfbba..5c5e72d304069535e7ea2e73584def0e4ce45156 100644
GIT binary patch
delta 4132
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delta 4131
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diff --git a/tests/reference_outputs/testdata/results/seurat_filtered/filtered_seurat_tenx_test.rds b/tests/reference_outputs/testdata/results/seurat_filtered/filtered_seurat_tenx_test.rds
index b7224f7815a0eaef0745007866191626a259f48d..99d55163955cc49a112baf63b25fdf103a8ff0eb 100644
GIT binary patch
delta 1799
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zOuk{u;XsnDj(Ru&oJ7mL6A95puTe`0?d*oi?r^tq4O|Gy?E7!Vmip!PFw|qnaD#+<
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zjQg0?GXK4_EW6b4VL`KynCV6!%2%?B)--7|+$VUD0_I`$W0<i;>{J$`?s&~xQ||0B
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delta 1831
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diff --git a/tests/reference_outputs/testdata/results/soupx/seurat_soupx_emptydrops_test.rds b/tests/reference_outputs/testdata/results/soupx/seurat_soupx_emptydrops_test.rds
index d3403c03cf62647cadbbcf490963cc3983634553..2de8ec6bc8f7ed9743cb0eed1d847437322a71a9 100644
GIT binary patch
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diff --git a/workflow/rules/install_doubletfinder.smk b/workflow/rules/install_doubletfinder.smk
index 4c5b95c..dc4bf7f 100644
--- a/workflow/rules/install_doubletfinder.smk
+++ b/workflow/rules/install_doubletfinder.smk
@@ -9,6 +9,6 @@ rule install_doubletfinder:
         "../envs/doubletfinder.yml"
     shell:
         """
-        Rscript -e "remotes::install_github('chris-mcginnis-ucsf/DoubletFinder', force = TRUE)" > {log} 2>&1
+        Rscript -e 'remotes::install_github("chris-mcginnis-ucsf/DoubletFinder", force = TRUE, dependencies = FALSE, upgrade = "never"); stopifnot(requireNamespace("DoubletFinder", quietly = TRUE), requireNamespace("Seurat", quietly = TRUE), requireNamespace("igraph", quietly = TRUE))' > {log} 2>&1
         touch {output}
         """

From 2da1944ebcb427932610b3ec23f1772093581acc Mon Sep 17 00:00:00 2001
From: Adam Freedman <adamfreedman@fas.harvard.edu>
Date: Thu, 11 Jun 2026 16:45:16 -0400
Subject: [PATCH 6/6] Relax neighborhood purity metadata tolerance

---
 tests/README.md                 | 2 +-
 tests/compare_seurat_metadata.R | 2 +-
 2 files changed, 2 insertions(+), 2 deletions(-)

diff --git a/tests/README.md b/tests/README.md
index 691bb75..35218ba 100644
--- a/tests/README.md
+++ b/tests/README.md
@@ -57,7 +57,7 @@ The GitHub-hosted full workflow action uses `tests/bin/cellbender` as a test shi
 
 ## Reference outputs
 
-The full workflow test compares regenerated files in `testdata/results/` against reference files under `tests/reference_outputs/`. The compared file list is in `tests/test_reference_output_files.txt`. Seurat `.rds` files are compared at the metadata-table level, marker CSVs are compared by columns and `(cluster, genesymbol)` rows with numeric tolerance, emptyDrops matrix files are compared after gzip decompression, CellBender H5 outputs are compared with `h5diff`, and remaining durable outputs are compared byte-for-byte. Seurat metadata numeric columns use a strict default tolerance, except `neighborhood_purity`, which allows a small absolute tolerance because it is a derived nearest-neighbor purity metric that can vary slightly across R/Bioconductor/platform builds even when barcode order and `seurat_clusters` match. Override that tolerance with `SEURAT_METADATA_NEIGHBORHOOD_PURITY_TOLERANCE` if needed.
+The full workflow test compares regenerated files in `testdata/results/` against reference files under `tests/reference_outputs/`. The compared file list is in `tests/test_reference_output_files.txt`. Seurat `.rds` files are compared at the metadata-table level, marker CSVs are compared by columns and `(cluster, genesymbol)` rows with numeric tolerance, emptyDrops matrix files are compared after gzip decompression, CellBender H5 outputs are compared with `h5diff`, and remaining durable outputs are compared byte-for-byte. Seurat metadata numeric columns use a strict default tolerance, except `neighborhood_purity`, which allows an absolute tolerance of `0.025` because it is a derived nearest-neighbor purity metric that can vary slightly across R/Bioconductor/platform builds even when barcode order and `seurat_clusters` match. Override that tolerance with `SEURAT_METADATA_NEIGHBORHOOD_PURITY_TOLERANCE` if needed.
 
 To refresh the reference snapshot after intentionally changing workflow behavior, first run the full test workflow so `testdata/results/` contains the desired outputs, then run:
 
diff --git a/tests/compare_seurat_metadata.R b/tests/compare_seurat_metadata.R
index 249c58b..95f39ac 100644
--- a/tests/compare_seurat_metadata.R
+++ b/tests/compare_seurat_metadata.R
@@ -10,7 +10,7 @@ if (is.na(numeric_tolerance) || length(numeric_tolerance) != 1 || numeric_tolera
   stop("SEURAT_METADATA_NUMERIC_TOLERANCE must be a non-negative number", call. = FALSE)
 }
 
-neighborhood_purity_tolerance <- as.numeric(Sys.getenv("SEURAT_METADATA_NEIGHBORHOOD_PURITY_TOLERANCE", "0.02"))
+neighborhood_purity_tolerance <- as.numeric(Sys.getenv("SEURAT_METADATA_NEIGHBORHOOD_PURITY_TOLERANCE", "0.025"))
 if (is.na(neighborhood_purity_tolerance) || length(neighborhood_purity_tolerance) != 1 || neighborhood_purity_tolerance < 0) {
   stop("SEURAT_METADATA_NEIGHBORHOOD_PURITY_TOLERANCE must be a non-negative number", call. = FALSE)
 }