the code used for the preprint Embedding-based Instance Segmentation of Microscopy Images.

Overview

EmbedSeg

Introduction

This repository hosts the version of the code used for the preprint Embedding-based Instance Segmentation of Microscopy Images. For a short summary of the main attributes of the publication, please check out the project webpage.

We refer to the techniques elaborated in the publication, here as EmbedSeg. EmbedSeg is a method to perform instance-segmentation of objects in microscopy images, based on the ideas by Neven et al, 2019.

teaser

With EmbedSeg, we obtain state-of-the-art results on multiple real-world microscopy datasets. EmbedSeg has a small enough memory footprint (between 0.7 to about 3 GB) to allow network training on virtually all CUDA enabled hardware, including laptops.

Citation

If you find our work useful in your research, please consider citing:

@misc{lalit2021embeddingbased,
      title={Embedding-based Instance Segmentation of Microscopy Images}, 
      author={Manan Lalit and Pavel Tomancak and Florian Jug},
      year={2021},
      eprint={2101.10033},
      archivePrefix={arXiv},
      primaryClass={eess.IV}
}

Dependencies

We have tested this implementation using pytorch version 1.1.0 and cudatoolkit version 10.0 on a linux OS machine.

In order to replicate results mentioned in the publication, one could use the same virtual environment (EmbedSeg_environment.yml) as used by us. Create a new environment, for example, by entering the python command in the terminal conda env create -f path/to/EmbedSeg_environment.yml.

Getting Started

Please open a new terminal window and run the following commands one after the other.

git clone https://github.com/juglab/EmbedSeg.git
cd EmbedSeg
conda env create -f EmbedSeg_environment.yml
conda activate EmbedSegEnv
python3 -m pip install -e .
python3 -m ipykernel install --user --name EmbedSegEnv --display-name "EmbedSegEnv"
cd examples
jupyter notebook

(In case conda activate EmbedSegEnv generates an error, please try source activate EmbedSegEnv instead). Next, look in the examples directory, and try out the dsb-2018 example set of notebooks (to begin with). Please make sure to select Kernel > Change kernel to EmbedSegEnv.

Training & Inference on your data

*.tif-type images and the corresponding masks should be respectively present under images and masks, under directories train, val and test. (In order to prepare such instance masks, one could use the Fiji plugin Labkit as detailed here). These are cropped in smaller patches in the notebook 01-data.ipynb. The following would be a desired structure as to how data should be prepared.

$data_dir
└───$project-name
    |───train
        └───images
            └───X0.tif
            └───...
            └───Xn.tif
        └───masks
            └───Y0.tif
            └───...
            └───Yn.tif
    |───val
        └───images
            └───...
        └───masks
            └───...
    |───test
        └───images
            └───...
        └───masks
            └───...
Comments
  • How can I reduce memory for inference

    How can I reduce memory for inference

    Hi.

    I tried separately ran the notebook[bbbc010-2012] for inference provided by this repo but I had a memory allocation issue. I used batch size as 1.

    Is there any other parameters to reduce memory requirement?

    Also I set normalization_factor = 32767 if data_type=='8-bit' else 255 instead of normalization_factor = 65535 if data_type=='16-bit' else 255.

    But nothing changed.

    bug 
    opened by r-matsuzaka 9
  • [BUG]RuntimeError: result type Byte can't be cast to the desired output type Bool

    [BUG]RuntimeError: result type Byte can't be cast to the desired output type Bool

    Hi, again..

    When I run begin_evaluating(test_configs, verbose = False, avg_bg= avg_bg/normalization_factor) at predict notebook, I got the following error:

    2-D `test` dataloader created! Accessing data from ../../../data/bbbc010-2012/test/
    Number of images in `test` directory is 50
    Number of instances in `test` directory is 50
    Number of center images in `test` directory is 0
    *************************
    Creating branched erfnet with [4, 1] classes
    

    0%| | 0/50 [00:01<?, ?it/s]


    RuntimeError Traceback (most recent call last) /tmp/ipykernel_33/4185926816.py in ----> 1 begin_evaluating(test_configs, verbose = False, avg_bg= avg_bg/normalization_factor)

    /kaggle/input/embedsegv1/EmbedSeg/test.py in begin_evaluating(test_configs, verbose, mask_region, mask_intensity, avg_bg) 62 test(verbose = verbose, grid_x = test_configs['grid_x'], grid_y = test_configs['grid_y'], 63 pixel_x = test_configs['pixel_x'], pixel_y = test_configs['pixel_y'], ---> 64 one_hot = test_configs['dataset']['kwargs']['one_hot'], avg_bg = avg_bg, n_sigma=n_sigma) 65 elif(test_configs['name']=='3d'): 66 test_3d(verbose=verbose,

    /kaggle/input/embedsegv1/EmbedSeg/test.py in test(verbose, grid_y, grid_x, pixel_y, pixel_x, one_hot, avg_bg, n_sigma) 126 127 center_x, center_y, samples_x, samples_y, sample_spatial_embedding_x, sample_spatial_embedding_y, sigma_x, sigma_y,
    --> 128 color_sample_dic, color_embedding_dic = prepare_embedding_for_test_image(instance_map = instance_map, output = output, grid_x = grid_x, grid_y = grid_y, pixel_x = pixel_x, pixel_y =pixel_y, predictions =predictions, n_sigma = n_sigma) 129 130 base, _ = os.path.splitext(os.path.basename(sample['im_name'][0]))

    /kaggle/input/embedsegv1/EmbedSeg/utils/utils.py in prepare_embedding_for_test_image(instance_map, output, grid_x, grid_y, pixel_x, pixel_y, predictions, n_sigma) 483 sample_spatial_embedding_y[id.item()] = add_samples(samples_spatial_embeddings, 1, grid_y - 1, pixel_y) 484 center_image = predictions[id.item() - 1]['center-image'] # predictions is a list! --> 485 center_mask = in_mask & center_image.byte() 486 487

    RuntimeError: result type Byte can't be cast to the desired output type Bool

    bug 
    opened by r-matsuzaka 5
  • dsb-2018/01-data.ipynb ValueError: numpy.ndarray size changed, may indicate binary incompatibility. Expected 88 from C header, got 80 from PyObject

    dsb-2018/01-data.ipynb ValueError: numpy.ndarray size changed, may indicate binary incompatibility. Expected 88 from C header, got 80 from PyObject

    Hi,

    I am trying to run the firs example notebook, and I am failing at the very first cell...

    miniconda installation, creating the environment from your directions.

    conda env create -f EmbedSeg_environment.yml
    conda activate EmbedSegEnv
    python3 -m pip install -e .
    python3 -m ipykernel install --sys-prefix  --name EmbedSegEnv --display-name "EmbedSegEnv"
    

    (instead of --user to install it into the virtualenv instead of $HOME/.local)

    (EmbedSegEnv) [[email protected] EmbedSeg]$ pip3 list |grep numpy
    numpy                             1.19.4
    (EmbedSegEnv) [[email protected] EmbedSeg]$ pip3 list |grep hdm
    hdmedians                         0.14.1
    
    from tqdm import tqdm
    
    from glob import glob
    
    import tifffile
    
    import numpy as np
    
    import os
    
    from EmbedSeg.utils.preprocess_data import extract_data, split_train_val
    
    from EmbedSeg.utils.generate_crops import *
    
    ---------------------------------------------------------------------------
    ValueError                                Traceback (most recent call last)
    <ipython-input-1-54e5f42b447e> in <module>
          5 import os
          6 from EmbedSeg.utils.preprocess_data import extract_data, split_train_val
    ----> 7 from EmbedSeg.utils.generate_crops import *
    
    ~/git/github/juglab/EmbedSeg/EmbedSeg/utils/generate_crops.py in <module>
          5 from scipy.ndimage.morphology import binary_fill_holes
          6 from scipy.spatial import distance_matrix
    ----> 7 import hdmedians as hd
          8 from numba import jit
          9 
    
    /c7/home/tru/miniconda3/envs/EmbedSegEnv/lib/python3.7/site-packages/hdmedians/__init__.py in <module>
          4 
          5 from .medoid import medoid, nanmedoid
    ----> 6 from .geomedian import geomedian, nangeomedian
    
    hdmedians/geomedian.pyx in init hdmedians.geomedian()
    
    ValueError: numpy.ndarray size changed, may indicate binary incompatibility. Expected 88 from C header, got 80 from PyObject
    
    
    opened by truatpasteurdotfr 5
  • Where is cmap_60.npy?

    Where is cmap_60.npy?

    Hello again.

    I have a question about your elaborate notebook. I get stacked one section when loading cmap_60.npy.

    When I tried to load it, I got FileNotFoundError: [Errno 2] No such file or directory: '../../../cmaps/cmap_60.npy'.

    How can I prepare it?

    bug 
    opened by r-matsuzaka 4
  • Where is medoid used?

    Where is medoid used?

    Hi.

    I have a question about implementation about medoid which is mentioned in the paper. I found the calculation of it is done at https://github.com/juglab/EmbedSeg/blob/50f23233cf9564ff443c67c45a611ce665571c12/EmbedSeg/utils/generate_crops.py#L84

    But I could not found the clue that this funtion is called from any other python scripts.

    Could you tell me how the medoid is used in the code?

    bug 
    opened by r-matsuzaka 3
  • creating prediction without having val files

    creating prediction without having val files

    Hi, I am trying to create/generate prediction (part 3) but my dataset lacks validation files which prevent me from going further. I was wondering is there a specific function or code that can be implemented to tackle the issue or by default, validation files are required to generate prediction?

    opened by aminrezaei-img 2
  • License and general questions

    License and general questions

    embedseg seems promising,

    • why not use bsd or apache for license
    • how does embedseg compares to DenoiSeg in segmenting connected components, performance, efficiency, etc...
    opened by seekingdeep 2
  • [BUG] `workers` Parameter not Respected by DataLoaders

    [BUG] `workers` Parameter not Respected by DataLoaders

    Describe the bug Only 1 thread (core) is used for the dataloaders.

    To Reproduce Steps to reproduce the behavior:

    1. Spin up any of the training examples
    2. Set batch_size to something respectable, like 512
    3. Adjust workers dataloader parameter
    4. Examine CPU utilization

    Expected behavior Multiple cores get engaged and are used to feed the GPU(s).

    Screenshots Only 1 CPU Core Engaged

    Desktop (please complete the following information):

    • OS: Ubuntu 20.04.2 LTS
    • Graphics 2x GeForce GTX 3090

    Additional context

    train_dataset_dict = create_dataset_dict(
    	data_dir = data_dir, 
    	project_name = project_name,  
    	center = center, 
    	size = train_size, 
    	batch_size = train_batch_size, 
    	virtual_batch_multiplier = virtual_train_batch_multiplier, 
    	normalization_factor= normalization_factor,
    	one_hot = one_hot,
    	workers=16,
    	type = 'train'
    )
    

    To help debug, from the same virtual environment I put together this dummy script:

    import random
    import numpy as np
    from torch.utils.data import Dataset
    import torch
    from tqdm.auto import tqdm
    
    class TestDS(Dataset):
        def __len__(self):
            return 5000
    
        def __getitem__(self, index):
            z = np.zeros((256*256))
            for i in range(256*256): z[i] = i
            return z
            
    
    val_dataset = TestDS()
    val_dataset_it = torch.utils.data.DataLoader(
        val_dataset,
        batch_size=32,
        shuffle=True,
        drop_last=True,
        num_workers=12,
        pin_memory=True
    )
    
    while True:
        for i, sample in enumerate(tqdm(val_dataset_it)):
            sample = sample.to('cuda:1')
    

    Running the above results in proper core utilization: Cores Properly Engaged

    Even adding the following code at the head of EmbSeg training script does not help:

    import os
    os.environ["MKL_NUM_THREADS"] = "20"
    os.environ["OMP_NUM_THREADS"] = "20"
    
    bug 
    opened by authman 1
  • V0.2.5 - tag (d)

    V0.2.5 - tag (d)

    • Add Arabidopsis-Cells-CAM notebooks
    • Add stitch_2d and stitch_3d functions
    • Introduce num_workers while creating test_configs_dict
    • Correct path to labkit wiki
    opened by lmanan 0
  • V0.2.5 - tag (b)

    V0.2.5 - tag (b)

    • Fix resume path
    • Add updated docstrings
    • Hide display tags, save_images and virtual_batch_multiplier from 2d notebooks
    • Set drop_last=False while creating val_dataset_it (this helps if number of val crops is less than val_batch_size)
    opened by lmanan 0
  • v0.2.5 - tag (a)

    v0.2.5 - tag (a)

    • Make min-max-percentile normalization default
    • Update README
    • Better Visualization of crops and model predictions
    • Reduce text in train notebooks
    • Take away virtual_batch_multiplier as a user-defined attribute
    opened by lmanan 0
  • Pretrained models not found

    Pretrained models not found

    Hello,

    I found your links of pretrained models in this project page are 404. Do they still available? I want to try your models on our private dataset of 3D nuclei instance segmentation.

    Thank you! Best wishes.

    bug 
    opened by Chrisa142857 0
  • cublas Run time error

    cublas Run time error

    Describe the bug I am trying the example notebooks and successfully ran 01-data However, when I try the training notebook and being training the model, it takes a long time to initialise and then I get the following error: cublas runtime error : the GPU program failed to execute at C:/w/1/s/tmp_conda_3.7_044431/conda/conda-bld/pytorch_1556686009173/work/aten/src/THC/THCBlas.cu:259

    Desktop (please complete the following information):

    • OS: Tried this on Window 10 and Windows 11
    • Graphics NVIDIA RTX 3080

    Additional context Not sure if its a compatibility issue with RTX 30 series cards. I found a similar error for RTX 2080 cards on older pytorch https://github.com/pytorch/pytorch/issues/17334

    bug 
    opened by pr4deepr 1
  • TypeError: forward() missing 4 required positional arguments: 'prediction', 'instances', 'labels', and 'center_images'[BUG]

    TypeError: forward() missing 4 required positional arguments: 'prediction', 'instances', 'labels', and 'center_images'[BUG]

    Hello. I tried tutorial of bbbc010-2012 Jupyter notebooks, but this error happend and I don't know solution. Could you tell me what I should do ?

    I ran 01-data.ipynb and 02-train.ipynb. When I ran 「begin_training(train_dataset_dict, val_dataset_dict, model_dict, loss_dict, configs, color_map=new_cmap)」, the following error happend. image image

    Environment

    • OS: Ubuntu 18.04
    • GPU:Tesla -python3.7 torch 1.1.0 torchvision 0.3.0 cuda=10.0
    bug 
    opened by kenta-takizawa 2
  • RuntimeError: CUDA out of memory.

    RuntimeError: CUDA out of memory.

    I have 4 images, and batch size is only 1. but when I start the begin_training(train_dataset_dict, val_dataset_dict, model_dict, loss_dict, configs), I have RuntimeError: CUDA out of memory. Tried to allocate 128.00 MiB (GPU 0; 31.75 GiB total capacity; 30.71 GiB already allocated; 62.50 MiB free; 12.93 MiB cached). Please let me know how can I solve it. Thanks

    opened by Saharkakavand 8
Releases(v0.2.4-tag)
  • v0.2.4-tag(Apr 18, 2022)

    This release was used to compute numbers for the MIDL publication and is stable.

    • The normalization of the image intensities was done by dividing pixel intensities by 255 (for 8-bit images) and 65535 (for unsigned 16-bit images). While this normalization strategy lead to a faster training, it lead to a sometimes, poorer OOD performance. In the future releases, the default will be set to min-max-percentile (takes model longer to reach the same val IoU but leads to a better inference performance).
    Source code(tar.gz)
    Source code(zip)
  • v0.2.3-tag(Jun 15, 2021)

    A minor update since release v0.2.2. This includes:

    • Add display_zslice parameter and save_checkpoint_frequency parameter to configs dictionary here
    1. Support for visualization for setups when virtual_batch_multiplier > 1 is still missing.
    2. Also hardcoded install version of tifffile in setup.py here because latest version currently (2021.6.14) generates a warning message with imsave command while generating crops with bbbc010-2012 dataset. Will relax this version specification in release v0.2.4

    TODOs include:

    1. Plan to update pytorch version to 1.9.0 in release v0.2.4 (currently pytorch version used is 1.1.0)
    2. Plan to add tile and stitch capability in release v0.2.4 for handling in large 2d and 3d images during inference
    3. Plan to add a parameter max_crops_per_image in release v0.2.4 to set an optional upper bound on number of crops extracted from each image
    4. Plan to save all instance crops and center crops as RLE files in release v0.2.4
    5. Plan to add an optional mask parameter during training which ignores loss computation from certain regions of the image in release v0.2.4
    6. Plan to deal with bug while evaluating var_loss and to have crops of desired size by additional padding.
    7. Plan to include support for more classes.
    8. Normalization for 3d ==> (0,1, 2)
    9. Make normalization as default option for better extensibility
    10. Parallelize operations like cropping
    11. Eliminate the specification of grid size in notebooks -set to some default value
    12. Simplify notebooks further
    13. Make colab versions of the notebooks
    14. Test center=learn capability for learning the center freely
    15. Add the ILP formulation for stitching 2d instance predictions
    16. Add the code for converting predictions from 2d model on xy, yz and xz slices to generate a 3D instance segmentation
    17. Add more examples from medical image datasets
    18. Add threejs visualizations of the instance segmentations. Explain how to generate these meshes, smoothen them and import them with threejs script.
    19. Padding with reflection instead of constant mode
    20. Include cluster_with_seeds in case nuclei or cell detections are additionally available
    Source code(tar.gz)
    Source code(zip)
  • v0.2.2-tag(May 5, 2021)

  • v0.2.0(Apr 17, 2021)

    Major changes:

    • Add 3d example notebooks for two datasets
    • Correct min_object_size (evaluated now from looking at the train and validation masks)
    • Save tif images with datatype np.uint16 (in the prediction notebooks )
    • Provide support in case evaluation GT images are not available (during prediction)

    Some things which are still incorrect in v0.2.0:

    • n_y should be set to n_x for equal pixel/voxel sizes in y and x dimension. This is fixed in v0.2.1
    • anisotropy_factor is wrongly calculated for the 3d notebooks (it was calculated as the reciprocal). This is fixed in v0.2.1
    • train_size was set to 600 for the bbbc012-2010 dataset. This is raised to 1200 in v0.2.1
    Source code(tar.gz)
    Source code(zip)
Owner
JugLab
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JugLab
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