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Final major update before merge
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fercrcode committed May 28, 2021
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36 changes: 22 additions & 14 deletions README.md
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Expand Up @@ -41,7 +41,8 @@ Overview of CyGNAL (dashed blue line) within a standard mass cytometry analysis:

## 1. System requirements

OS info here
CyGNAL has been tested on both macOS (from Catalina onwards) and Debian-based
Linux distributions (including Ubuntu on [WSL](https://github.com/Microsoft/WSL)).

### Dependencies

Expand All @@ -58,14 +59,15 @@ workflow and most computational steps.
* `sklearn`
* `umap-learn`

* R: Tested with v3.6.1 < R <= v4. Mostly used for visualisation, but also for
* R: Tested with v3.6 < R <= v4.0. Mostly used for visualisation, but also for
computing the PCA.
* `ComplexHeatmap`
* `DT`
* `factoextra`
* `FactoMineR`
* `flowCore`
* `Ggally`
* `ggrepel`ma
* `Hmisc`
* `MASS`
* `matrixStats`
Expand All @@ -81,16 +83,16 @@ computing the PCA.

## 2. Using CyGNAL

CyGNAL is distributed as a multilevel directory. The 'code' folder contains the
main steps, with other utility scripts found in 'code/utils/'.
Input data should be added to 'Raw_Data' for pre-processing and processed
CyGNAL is distributed as a set of directories. The 'code' folder contains the
main steps, with other utility scripts found in 'code/utils/', to be run as `python` scripts.
Input data should be added to 'Raw_Data' for pre-processing, and processed
datasets are stored in 'Preprocessed_Data'. Input and output directories for
the analysis and visualisation steps are found in the 'Analysis' directory.

### Input data

Raw data contains sample dataset files. Pipeline can take in both FCS and
.txt files (as tab-separated dataframes).
CyGNAL can take in both FCS and .txt files (as tab-separated dataframes and
without a header). The 'Raw Data' directory contains sample dataset files.

*NOTE*: The toy dataset used in this tutorial is a down-sampled version
(5,000 cells per time point, EpCAM/Pan-CK gated) of the small intestinal
Expand Down Expand Up @@ -129,7 +131,8 @@ file listing all the markers measured in the given experiment.
population(s) of interest, and export events as untransformed text files
(Actions - Export - Export events, with *'Include header with FCS filename'* unchecked).

*Note:* This step is essential for getting the dataset compatible with downstream analysis and has to be performed as the first step in our workflow.
*Note:* This step is essential for getting the dataset compatible with
downstream analysis and has to be performed as the first step in our workflow.

2. **UMAP:** Move the processed data file(s) and panel_marker.csv to 'Analysis/UMAP_input'.
Edit *'panel_markers.csv'* to set all the markers used for UMAP analysis from 'N' to 'Y'.
Expand All @@ -146,11 +149,11 @@ The markers and the indices of the cells used in the analysis will also be saved
3. **EMD:** To perform EMD calculation (using the tools available in the
[scprep](https://github.com/KrishnaswamyLab/scprep) library), copy the input
data files to 'Analysis/EMD_input'. Run `3-emd.py` and follow the instructions.
By default, the denominator of the EMD calculation will be the concatenation
By default, the reference of the EMD calculation will be the concatenation
of all the input data files, but the user is given the option to provide a
specific denominator data file. While EMD scores of all channels can be
calculated by default, by default the user should place the *'panel_markers.csv'*
in the input folder to specifiy which marker are to be used.
specific reference data file. While EMD scores of all channels can be
calculated, the default behaviour requires the user to place the *'panel_markers.csv'*
in the input folder to specifiy which markers are to be used.
The calculated EMD scores will be saved in 'Analysis/EMD_output', within the
'EMD_arc_no_norm' column in the saved file.
* `python 3-emd.py`
Expand Down Expand Up @@ -190,12 +193,17 @@ Ferran Cardoso ([@FerranC96](https://github.com/FerranC96)) and
Dr. Xiao Qin ([@qinxiao1990](https://github.com/qinxiao1990)).
Based also on original work by Pelagia Kyriakidou.

We acknowledge the work of all third-parties whose packages are used in CyGNAL.
### Support

For any queries or issues regarding CyGNAL please check the
[Issues](https://github.com/TAPE-Lab/CyGNAL/issues) section in this repository.

### The group

Repository of the [Cell Communication Lab](http://tape-lab.com/) at UCL's Cancer Institute.
The Cell Communication Lab studies how oncogenic mutations communicate with
stromal and immune cells in the colorectal cancer (CRC) tumour microenvironment (TME).
By understanding how mutations regulate all cell types within a tumour,
we aim to uncover novel approaches to treat cancer.
we aim to uncover novel approaches to treat cancer.

We acknowledge the work of all third-parties whose packages are used in CyGNAL.
2 changes: 1 addition & 1 deletion conda_env.yml
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Expand Up @@ -30,7 +30,7 @@ dependencies:
- r-ggrepel #in conda-forge
- bioconda::bioconductor-complexheatmap #noarch
- bioconda::bioconductor-flowcore=2.2.0
- bioconda::bioconductor-cytolib=2.2.1 #2.2.1 fixed on 01/05/21
- bioconda::bioconductor-cytolib=2.2.1 #2.2.1 set on 01/05/21
- pip:
- fcswrite

1 change: 1 addition & 0 deletions dependency_troubleshoot.py
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Expand Up @@ -30,6 +30,7 @@
"FactoMineR",
"flowCore",
"GGally",
"ggrepel",
"Hmisc",
"MASS",
"matrixStats",
Expand Down

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