toomanycells 1.0.2

A python package for spectral clustering.

too-many-cells (à la Python)

It's Scanpy friendly!

**A python package for spectral clustering based on the powerful suite of tools named too-many-cells. In essence, you can use toomanycells to partition a data set in the form of a matrix of integers or floating point numbers into clusters. The rows represent observations and the columns are the features. Initially, toomanycells will partition your data set into two subsets, trying to maximize the differences between the two. Subsequently, it will reapply that same criterion to each subset and will continue bifurcating until the modularity of the parent becomes negative, implying that the current subset is fairly homogeneous, and consequently suggesting that further partitioning is not warranted. Thus, when the process finishes, you end up with a tree structure of your data set, where the leaves represent the clusters. As mentioned earlier, you can use this tool with any kind of data. However, a common application is to classify cells and therefore you can provide an AnnData object. You can read about this application in this Nature Methods paper. **

• Free software: BSD 3-Clause License
• Documentation: https://JRR3.github.io/toomanycells

Dependencies

Make sure you have installed the graph visualization library Graphviz.

Installation

Just type

pip install toomanycells

in your home environment.

Usage

1. First import the module as follows

   from toomanycells import TooManyCells as tmc
   

2. If you already have an AnnData object A loaded into memory, then you can create a TooManyCells object with

   tmc_obj = tmc(A)
   

   However, if you want the output folder to be a directory that is not the current working directory, then you can specify the path as follows

   tmc_obj = tmc(A, output_directory)
   

3. If instead of providing an AnnData object you want to provide the directory where your data is located, you can use the syntax

   tmc_obj = tmc(input_directory, output_directory)
   

4. If your input directory has a file in the matrix market format, then you have to specify this information by using the following flag

   tmc_obj = tmc(input_directory, output_directory, input_is_matrix_market=True)
   

   Under this scenario, the input_directory must contain a .mtx file, a barcodes.tsv file (the observations), and a genes.tsv (the features).
5. Once your data has been loaded successfully, you can start the clustering process with the following command

   tmc_obj.run_spectral_clustering()
   

   In my desktop computer a data set with ~90K cells (observations) and ~30K genes (features) it took a little less than 6 minutes in 1809 iterations. Note that the progress bar only produces an estimate of the total number of iterations.
6. At the end of the clustering process the .obs data frame of the AnnData object should have two columns named ['sp_cluster', 'sp_path'] which contain the cluster labels and the path from the root node to the leaf node, respectively.

   tmc_obj.A.obs[['sp_cluster', 'sp_path']]
   

7. To generate the outputs, just call the function

   tmc_obj.store_outputs()
   

   This call will generate a PDF of the tree and a DOT file for the graph, two CSV files that describe the clusters and the information of each node, and a JSON file that contains the tree structure. If you already have a DOT file and only want to plot the tree and store the information of each node, you can use the following call

   tmc_obj.store_outputs(load_dot_file=True)
   

8. If you want to visualize your results in a dynamic platform, I strongly recommend the tool too-many-cells-interactive. To use it, first make sure that you have Docker Compose and Docker. One simple way of getting the two is by installing Docker Desktop. If you use Nix, simply add the packages pkgs.docker and pkgs.docker-compose to your configuration or home.nix file and run

home-manager switch

9. If you installed Docker Desktop you probably don't need to follow this step. However, under some distributions the following two commands have proven to be essential.

sudo dockerd

to start the daemon service for docker containers and

sudo chmod 666 /var/run/docker.sock

to let Docker read and write to that location.

10. Now clone the repository

git clone https://github.com/schwartzlab-methods/too-many-cells-interactive.git

and store the path to the too-many-cells-interactive folder in a variable, for example path_to_tmc_interactive. Also, you will need to identify a column in your AnnData.obs data frame that has the labels for the cells. Let's assume that the column name is stored in the variable cell_annotations. Lastly, you can provide a port number to host your visualization, for instance port_id=1234. Then, you can call the function

tmc_obj.visualize_with_tmc_interactive(
         path_to_tmc_interactive,
         cell_annotations,
         port_id)

In my case it took a little less than 3 minutes to load the app with ~90K cells (observations).