Van Dijk, David, et al. "Recovering Gene Interactions from Single-Cell Data Using Data Diffusion." Cell (2018).
https://www.ncbi.nlm.nih.gov/pubmed/29961576
MAGIC has been implemented in Python3 and Matlab. See below for installation and Usage.
MAGIC is an unsupervised non-parametric algorithm to impyte and de-noise biological single-cell RNA-seq data sets. MAGIC achieves this objective by sharing information across similar cells via data diffusion. Algorithmically, MAGIC performs the following operations in the sequential order:
1). Constructs a nearest-neighbor graph
2). Converts the distance matrix to an affinity matrix, via a Gaussian kernel
3). Converts the affinity matrix to a Markov (or Transition) matrix, via row normalization
4). Raises the Markov matrix to a power t, to emulate data diffusion
5). The original data is then right-multiplied by the powered Markov matrix
MAGIC has a few important parameters that play important rolw to determine the quality of results. They are listed below:
1). k : This defines the number of nearest neighbors used to construct the graph. We recomment this to be small enough that only local neighborhood of each cell is considered but big enough that the graph remains connected. By default, this is set to k = 30.
2). ka : This dictates the standard deviation to be used in the Gaussian kernel. To elaborate, the standard deviation in the Gaussian kernel for a given cell is set to be the distance to it's ka-th nearest neighbor. By default, this is set to ka = k/3 = 10.
3). t : This defines the power to which the Markov matrix is to be raised. This is arguably the most important parameter. A very high t can lead to over-smoothed results while a low t can lead to noisy results. We provide an automatic way to detect t in the paper. For this, we compute the degree of change between the imputed data at time t and time t-1 and stop after this value stabilizes. With the increase in t, the data goes through a rapidly changing imputation regime followed by a smoothing regime. In the imputation regime, the diffusion learns the manifold structure and removes noise. At larger values of t, diffusing further would smooth out real biology. The knee-point determines an optimal t. However, the choice of t can be context-dependent. While the automatic method can act as a good guide to choose optimal t, the underlying mathematics may not always recapitulate the true biology. Therefore, we always recommend to spend time looking at the data and to vet the values of t accordingly. For more details on the method please view run_magic.m file inside matlab directory above.
For example Python notebooks, please go inside the notebooks directory and open: Magic_single_cell_RNAseq.ipynb
For example MATLAb usage, please go inside the notebooks directory and open: test_magic.m
For a tutorial on how to use the GUI, please go inside docs and open: magic_tutorial.pptx
-
The Python3 version of MAGIC can be installed using:
$> git clone git://github.com/pkathail/magic.git $> cd magic $> sudo -H pip3 install . -
MAGIC depends on a number of
python3packages available on pypi and these dependencies are listed insetup.pyAll the dependencies will be automatically installed using the above commands -
After pulling updates to MAGIC from github, the package must be uninstalled and reinstalled:
$> sudo -H pip3 uninstall magic $> sudo -H pip3 install .
- run_magic.m -- MAGIC imputation function
- test_magic.m -- Shows how to run MAGIC. Also included is a function for loading 10x format data (load_10x.m)
A tutorial on MAGIC usage and results visualization for single cell RNA-seq data can be found in this notebook: http://nbviewer.jupyter.org/github/pkathail/magic/blob/develop/notebooks/Magic_single_cell_RNAseq.ipynb
A python GUI is now available for MAGIC. After following the installation steps listed below, the GUI can be invoked using
$> magic_gui.py
MAGIC can be run using the command line script MAGIC.py with the following parameters:
$> MAGIC.py -h
usage: MAGIC.py [-h] -d D -o O [-g G] [--gene-name-file GN]
[--use-ensemble-ids] [--cell-axis CA] [--skip-rows SKIP_ROWS]
[--skip-columns SKIP_COLUMNS] [-n] [-l L]
[--mols-per-cell-min MOLS_PER_CELL_MIN]
[--mols-per-cell-max MOLS_PER_CELL_MAX] [-p P]
[--pca-non-random] [-t T] [-k K] [-ka KA] [-e E] [-r R]
{csv,10x,10x_HDF5,mtx}
run MAGIC
positional arguments:
{csv,10x,mtx} what is the file type of your original data?
optional arguments:
-h, --help show this help message and exit
data loading parameters:
-d D, --data-file D File path of input data file.
-o O, --output-file O
File path of where to save the MAGIC imputed data (in
csv format).
-g G, --genome G Genome must be specified when loading 10x_HDF5 data.
--gene-name-file GN Gene name file must be specified when loading mtx
data.
--use-ensemble-ids Use ensemble IDs instead of gene names.
--cell-axis CA When loading a csv, specify whether cells are on rows
or columns (Default = 'rows').
--skip-rows SKIP_ROWS
When loading a csv, number of rows to skip after the
header row (Default = 0).
--skip-columns SKIP_COLUMNS
When loading a csv, number of columns to skip after
the header columns (Default = 0).
normalization/filtering parameters:
-n, --no-normalize Do not perform library size normalization on the data
-l L, --log-transform L
Log-transform data with the specified pseudocount.
--mols-per-cell-min MOLS_PER_CELL_MIN
Minimum molecules/cell to use in filtering.
--mols-per-cell-max MOLS_PER_CELL_MAX
Maximum molecules/cell to use in filtering.
MAGIC parameters:
-p P, --pca-components P
Number of pca components to use when running MAGIC
(Default = 20).
--pca-non-random Do not used randomized solver in PCA computation.
-t T t parameter for running MAGIC (Default = 6).
-k K Number of nearest neighbors to use when running MAGIC
(Default = 30).
-ka KA knn-autotune parameter for running MAGIC (Default =
10).
-e E, --epsilon E Epsilon parameter for running MAGIC (Default = 1).
-r R, --rescale R Percentile to rescale data to after running MAGIC
(Default = 99).
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