Xenium

Sample details

Study Name

High resolution mapping of the breast cancer tumor microenvironment using integrated single cell, spatial and in situ analysis of FFPE tissue

WebAtlas

Demo

Tissue

Human breast cancer

Data Source Link

Xenium_FFPE_Human_Breast_Cancer_Rep1_outs.zip

Steps to reproduce

Follow the steps below to reproduce this sample in the pipeline, and visualise the data yourself in your web browser. It can be followed on any POSIX compatible system (Linux, OS X, etc). This example requires you to have already setup your environment first.

1. Download the sample data

Using wget

Input
mkdir -p input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs
wget https://cf.10xgenomics.com/samples/xenium/1.0.1/Xenium_FFPE_Human_Breast_Cancer_Rep1/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs.zip -P input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs
Output
--2023-05-17 15:05:24--  https://cf.10xgenomics.com/samples/xenium/1.0.1/Xenium_FFPE_Human_Breast_Cancer_Rep1/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs.zip
Resolving cf.10xgenomics.com (cf.10xgenomics.com)... 104.18.0.173, 104.18.1.173, 2606:4700::6812:ad, ...
Connecting to cf.10xgenomics.com (cf.10xgenomics.com)|104.18.0.173|:443... connected.
HTTP request sent, awaiting response... 200 OK
Length: 9861155708 (9.2G) [application/zip]
Saving to: ‘input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs.zip’

Xenium_FFPE_Human_Breast_Cancer_Rep1 100%[===================================================================>]   9.18G  14.3MB/s    in 10m 6s

2023-05-17 15:15:31 (15.5 MB/s) - ‘input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs.zip’ saved [9861155708/9861155708]

Using curl

Input
mkdir -p input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs
curl -O --output-dir input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs https://cf.10xgenomics.com/samples/xenium/1.0.1/Xenium_FFPE_Human_Breast_Cancer_Rep1/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs.zip
Output
  % Total    % Received % Xferd  Average Speed   Time    Time     Time  Current
                                 Dload  Upload   Total   Spent    Left  Speed
100 9404M  100 9404M    0     0  9607k      0  0:16:42  0:16:42 --:--:--  9.8M

2. Extract the sample data

Input
unzip input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs.zip -d input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs
Output
Archive:  input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs.zip
   creating: input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/outs/
  inflating: input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/outs/experiment.xenium
   creating: input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/outs/cell_feature_matrix/
  inflating: input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/outs/cell_feature_matrix/barcodes.tsv.gz
         ...
         ...
  inflating: input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/outs/metrics_summary.csv
  inflating: input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/outs/gene_panel.json
  inflating: input/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/outs/analysis_summary.html

3. Run the pipeline

Warning

Users working on newer Silicon-based Macs may encounter problems with the conda environment. Some packages have not yet been compiled for Apple silicon processors therefore, we recommend you install the packages originally compiled for Mac computers with Intel processors. Set an environment variable that specifies the architecture before running the pipeline, like this:

Input
export CONDA_SUBDIR=osx-64
Input
nextflow run main.nf \
      -params-file templates/examples/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs.yaml \
      -entry Full_pipeline \
      -profile conda
Output
 N E X T F L O W  ~  version 22.10.6
 Launching `main.nf` [gigantic_murdock] DSL2 - revision: 1b6a73f4d6
 [fc/782a3f] process > Full_pipeline:Process_files:route_file (xenium, outs)                              [100%] 1 of 1  [b0/f5ff27] process > Full_pipeline:Process_images:Generate_image ([xenium, breast-cancer], label, outs) [100%] 1 of 1  [2b/054048] process > Full_pipeline:Process_images:image_to_zarr (morphology.ome.tif)                    [100%] 2 of 2  [07/5e37c4] process > Full_pipeline:Process_images:ome_zarr_metadata (METADATA.ome.xml)                  [100%] 2 of 2  [c8/f2378c] process > Full_pipeline:Output_to_config:Build_config ([xenium, breast-cancer])              [100%] 1 of 1  Completed at: 17-May-2023 16:40:58
 Duration    : 32m 47s
 CPU hours   : 0.6
 Succeeded   : 7

4. Check execution was successful

The output from the pipeline will indicate if the execution was successful. You can also verify the expected directories are created.

Input
ls -l output/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/0.5.2
Output
 total 3566252
 drwxrwxr-x 11 dh74 dh74       4096 May 17 16:08 xenium-breast-cancer-anndata.zarr
 -rw-r--r--  1 dh74 dh74       4984 May 17 16:40 xenium-breast-cancer-config.json
 -rw-r--r--  1 dh74 dh74 3651814848 May 17 16:12 xenium-breast-cancer-label.tif
 drwxrwxr-x  4 dh74 dh74       4096 May 17 16:13 xenium-breast-cancer-label.zarr
 drwxrwxr-x  4 dh74 dh74       4096 May 17 16:40 xenium-breast-cancer-raw.zarr

5. Serve the data output through a local web server

To browse and explore the data, you need to serve the output data through a web server. You can use your preferred web server, but you must ensure the data is served over port 3000, at http://localhost:3000, and that CORS is enabled via the Access-Control-Allow-Origin header.

Input
npx http-server output/Xenium_FFPE_Human_Breast_Cancer_Rep1_outs/0.5.2 --port 3000 --cors
Output
Starting up http-server, serving ./

http-server version: 14.1.1

http-server settings:
CORS: true
Cache: 3600 seconds
Connection Timeout: 120 seconds
Directory Listings: visible
AutoIndex: visible
Serve GZIP Files: false
Serve Brotli Files: false
Default File Extension: none

Available on:
http://127.0.0.1:3000
http://192.168.0.23:3000
Hit CTRL-C to stop the server

6. Explore data in your browser

Start your web browser and open:

https://webatlas.cog.sanger.ac.uk/latest/index.html?theme=dark&config=http://127.0.0.1:3000/xenium-breast-cancer-config.json