Light Beads Microscopy 2P Calcium Imaging Pipeline.
Project description
LBM-CaImAn-Python
Python implementation of the Light Beads Microscopy (LBM) computational pipeline.
For the MATLAB
implementation, see here
Pipeline Steps:
- Image Assembly
- Extract raw
tiffs
to planar timeseries
- Extract raw
- Motion Correction
- Template creation
- Piecewise-rigid or non-rigid registration
- Segmentation
- Iterative CNMF segmentation
- Deconvolution
- Refinement
- Collation
- Lateral offset correction (between z-planes)
- Collate images and metadata into a single volume
Requirements
- caiman
- mesmerize-core
- scanreader
- numpy
- scipy
- fastplotlib
:exclamation: Note: This package makes heavy use of Fastplotlib for visualizations.
Fastplotlib runs on Jupyter Lab, but is not guarenteed to work with Jupyter Notebook or Visual Studio Code notebook environments.
Installation
This project is tested on Linux and Windows 10 using Python 3.9
and Python 3.10
.
Environment setup is tested using virtualenv
and miniforge
.
We suggest using python virtual environments for the best results.
(Option 1). Python Virtual Environments
Ensure you have a system-wide Python installation.
This project and it's dependencies are tested using Python 3.9
and Python 3.10
.
Note: Make sure you deactivate conda
environments before proceeding (conda deactivate
).
Verify Python
and pip
installations:
- Linux/macOS:
python --version
pip --version
- Windows:
py --version
py - m pip --version
:exclamation: Depending on how Python was installed,
you may need to use python3
or python3.x
and pip3
or pip3.x
instead of python
and pip
.
You should see a Python version output like 3.10.x
and a corresponding pip
version.
If Python is not installed, or an unsupported version is installed (i.e. 3.7),
download and install python.org or refer to this installation guide.
You will also need git
:
git --version
Create a virtual environment
This is normally in a directory dedicated to virtual environments, but can be anywhere you wish:
python -m venv ~/venv/lbm_caiman_python
Activate the virtual environment:
-
Linux/macOS:
source ~/venv/lbm_caiman_python/bin/activate
-
Windows:
source ~/venv/lbm_caiman_python/Scripts/activate
Upgrade core tools in the virtual environment:
pip install --upgrade setuptools wheel pip
Clone and install CaImAn
Create a directory to store the cloned repositories.
Again, this can be anywhere you wish:
cd ~
mkdir repos
cd repos
Use git to clone CaImAn:
git clone https://github.com/flatironinstitute/CaImAn.git
Install CaImAn:
-
CaImAn:
cd CaImAn pip install -r requirements.txt pip install .
:exclamation: Note: If you encounter errors during the installation of
CaImAn
, you may need to install Microsoft Visual C++ Build Tools. You can download them from here. -
Other dependencies:
pip install mesmerize-core pip install lbm_caiman_python pip install git+https://github.com/atlab/scanreader.git
Run ipython to make sure everyting works
import lbm_caiman_python as lcp
import mesmerize_core as mc
import scanreader as sr
scan = sr.read_scan('path/to/data/*.tif', join_contiguous=True)
virtualenv Troubleshooting
Error During pip install .
(CaImAn) on Linux
If you encounter errors during the installation of CaImAn
, install the necessary development tools:
sudo apt-get install python3-dev
(Option 2). Conda
Miniforge is the supported conda
distribution. Anaconda and Miniconda require extra steps and is not covered in this guide.
Note: Sometimes conda or mamba will get stuck at a step, such as creating an environment or installing a package.
Pressing Enter on your keyboard can sometimes help it continue when it pauses.
- Install
mamba
into your base environment:
:exclamation: This step may take 10 minutes and display several messages like "Solving environment: failed with..." but it should eventually install mamba.
conda activate base
conda install -c conda-forge mamba
- Create a new environment and install mesmerize-core
- Here, we use the
-n
flag to name the environmentlbm
, but you can name it whatever you'd like. - This step will install Python, mesmerize-core, CaImAn, and all required dependencies for those packages.
conda create -n lbm -c conda-forge mesmerize-core
If you already have CaImAn
installed:
conda install -n name-of-env-with-caiman mesmerize-core
Activate the environment and install caimanmanager
:
- if you used a name other than
lbm
, be sure to match the name you use here.
conda activate lbm
caimanmanager install
- Install LBM-CaImAn-Python from pip:
pip install lbm_caiman_python
- Install scanreader:
pip install git+https://github.com/atlab/scanreader.git
- (Optional) Install
mesmerize-viz
:
Several notebooks make use of mesmerize-viz for visualizing registration/segmentation results.
pip install mesmerize-viz
:exclamation: Harware requirements The large CNMF visualizations with contours etc. usually require either a dedicated GPU or integrated GPU with access to at least 1GB of VRAM.
https://www.youtube.com/watch?v=GWvaEeqA1hw
For Developers
To get the newest version of this package:
git clone https://github.com/MillerBrainObservatory/LBM-CaImAn-Python.git
cd LBM-CaImAn-Python
pip install ".[docs]"
Troubleshooting
Conda Slow / Stalling
if conda is behaving slow, clean the conda installation and update conda-forge
:
conda clean -a
conda update -c conda-forge --all
Don't forget to press enter a few times if conda is taking a long time.
Recommended Conda Distribution
The recommended conda installer is miniforge.
This is a community-driven conda
/mamba
installer with pre-configured packages specific to conda-forge.
This helps avoid conda-channel
conflicts and avoids any issues with the Anaconda TOS.
You can install the installer from a unix command line:
curl -L -O "https://github.com/conda-forge/miniforge/releases/latest/download/Miniforge3-$(uname)-$(uname -m).sh"
bash Miniforge3-$(uname)-$(uname -m).sh
Or download the installer for your operating system here.
Graphics Driver Issues
If you are attempting to use fastplotlib and receive errors about graphics drivers, see the fastplotlib driver documentation.
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