Lumos

Lumos is a program that generates full-plate images from the separate site/field pictures obtained through the Cell Painting assay.

It has 2 operation modes that allow it to render images for specific channels of a plate, as well as colorful images that combine all channels together:

In this document, you will find information related to:

• The installation of the package
• How to use Lumos
• Detailed examples

To learn more about the command-line interface of the program, refer to the Wiki pages.

You can also find instructions for developers in this other documentation.

Installing Lumos

Prerequisites:

• You need to have Python 3.8 installed on a Windows or Linux machine.
• You also need to have the raw site images of your plates on your machine/mounted storage drive.

Then, to install Lumos, choose one of the following:

• Install Lumos using PIP with:

  pip install lumos-cli
  

• Install Lumos from the source files:
  • Download the source files, either by downloading the ZIP file or cloning the repository.
  • Install the dependencies using PIP: pip install -r .\requirements.txt
  • Install Lumos: python setup.py install

Usage

Configuring Lumos

To configure Lumos to work with your specific plate formats, you must first create a configuration file. To do so, enter the command below:

lumos --generate-config-file ./my_config.yaml

Then, in the generated file, you should change the following entries to match the organization of your images:

• path_from_run_folder_to_plates
• path_from_plate_folder_to_images
• input_file_naming_scheme
• well_grid
• site_grid
• image_dimensions

Then, also make sure that you update the following items with the correct ID of your channels (i.e. how they appear in your file names):

• the keys/sub-entries of channel_info
• default_channels_to_render
• brightfield_channels
• cp_channels_to_use

You can view an example of this here.

Once this is done, you are ready to use Lumos on your images.

Simple commands

A first command you can run is the following:

lumos --config-file ./my_config.yaml qc --scope plate --source-path <path_to_a_plate_folder> --output-path ./output/

where <path_to_a_plate_folder> is a path to the root folder of a plate. In this example, this would be one of the PLATE-XXX folders.

This will generate 1 greyscale image per channel of the plate (excluding brightfield channels) and allow you to check easily that the quality of the images that you have is good:

Note:

We could have also used argument aliases in order to make the command shorter, as follows:

lumos -cf ./my_config.yaml qc -s plate -sp <path_to_a_plate_folder> -op ./output/

To see a list of all available arguments and their aliases, use lumos qc --help.

This first command relied on the qc, or "Quality Control" mode of Lumos (hence the qc keyword present in the command). Similarly, Lumos also has a cp, or "Cell Painting" mode, that allows you to generate 1 combined image of all the channels of a plate (in color).

To use it, type the following in your terminal:

lumos -cf my_config.yaml cp --scope plate --source-path <path_to_a_plate_folder> --output-path ./output/

This will generate an image looking similar to this:

(zooming in: )

Note:

To see a list of all available Cell Painting arguments and their aliases, use lumos cp --help.

In addition to those first examples, Lumos QC can also generate images for only a specific channel of a plate, or for a whole run (i.e. a collection of plates). And similarly, Lumos CP can generate separate images for each well of a plate, just for a single well, or even for each site of those wells.

This, as well as other functionalities of Lumos, can be chosen through the arguments used in your commands. To see a full list of the available arguments and options you have for each operation mode, type lumos qc --help and lumos cp --help respectively.

You can also explore some of our more detailed examples or refer to our Wiki pages to learn more.

Detailed examples

Example configuration file

If you had the following folder structure:

JUMPCP_images/
    ├ run_1/
        └ plates/
            ├ PLATE-101/
              └ Images/
                  ├ r01c01f01p01-ch1sk1fk1fl1.tiff
                  ├ r01c01f01p01-ch2sk1fk1fl1.tiff
                  ├ ...
                  └ r32c48f04p01-ch8sk1fk1fl1.tiff
            ├ PLATE-102/
              └ Images/
                  ├ r01c01f01p01-ch1sk1fk1fl1.tiff
                  ├ r01c01f01p01-ch2sk1fk1fl1.tiff
                  ├ ...
                  └ r32c48f04p01-ch8sk1fk1fl1.tiff
            └ ...
    ├ run_2/
        └ plates/
            ├ PLATE-201/...
            ├ PLATE-202/...
            └ ...
    └ run_3/
        └ plates/
            ├ PLATE-301/...
            ├ PLATE-302/...
            └ ...

with wells in a grid of 32 rows by 48 columns, sites in a grid of 2 by 2, and images of 1080 by 1080 pixels.

You would edit the following entries in your configuration file as follows:

path_from_run_folder_to_plates: 'plates/'
path_from_plate_folder_to_images: 'Images/'
input_file_naming_scheme: rows_and_columns
well_grid: 32x48
site_grid: 2x2
image_dimensions: 1080x1080
channel_info:
  ch1: [...]
  ch2: [...]
  ch3: [...]
  ch4: [...]
  ch5: [...]
  ch6: [...]
  ch7: [...]
  ch8: [...]
default_channels_to_render:
- ch1
- ch2
- ch3
- ch4
- ch5
brightfield_channels:
- ch6
- ch7
- ch8
cp_channels_to_use:
- ch1
- ch2
- ch3
- ch4
- ch5

Example 1: Generating the image of a specific channel of a plate

To do this, we can use the 'channel' scope with the additional --channel argument to select which channel should be rendered by specifying its channel ID. Assuming the above example configuration, to render the channel "ch3" of our plate, we would get:

lumos --config-file my_config.yaml qc --scope channel --channel ch3 --source-path <path_to_a_plate_folder> --output-path ./output/

And alternatively, using argument aliases, we can use the shorter following command:

lumos -cf my_config.yaml qc -s channel -c ch3 -sp <path_to_a_plate_folder> -op ./output/

Example 2: Generating an image for all channels of a plate, including brightfields

To do this, we can add the --brightfield argument to our regular command. With this argument, we can either choose to include a specific Brightfield channel by writing its ID, or include all of them by writing 'all'. In this example, we will do the latter:

lumos --config-file my_config.yaml qc --scope plate --source-path <path_to_a_plate_folder> --output-path ./output/ --brightfield all

And alternatively, using argument aliases, we can use the shorter following command:

lumos -cf my_config.yaml qc -s plate -sp <path_to_a_plate_folder> -op ./output/ -b all

Example 3: Generating images for all channels of multiple plates (i.e. a "run")

To do this, we can use the 'run' scope. Then, instead of putting a path to a plate as our --source-path, we put the path to our "run" (i.e. our collection of plate).

lumos --config-file my_config.yaml qc --scope run --source-path <path_to_a_run_folder> --output-path ./output/

Note:

Make sure that your configuration file contains the correct path to follow between your run folder to your plate folders in path_from_run_folder_to_plates

And alternatively, using argument aliases, we can use the shorter following command:

lumos -cf my_config.yaml qc -s run -sp <path_to_a_run_folder> -op ./output/

Example 4: Generating the Cell Painted image of a specific well in a plate

This can be done using the 'wells' scope of the cp mode of Lumos. Then, we can restrict the generation of images to a single well using the --single-well argument followed by the well name (e.g. "r03c21"):

lumos --config-file my_config.yaml cp --scope wells --single-well r03c21 --source-path <path_to_a_plate_folder> --output-path ./output/

And alternatively, using argument aliases, we can use the shorter following command:

lumos -cf my_config.yaml cp -s wells -w r03c21 -sp <path_to_a_plate_folder> -op ./output/