U-FISH 🎣

U-FISH is an advanced FISH spot calling algorithm based on deep learning. The "U" in U-FISH represents both the U-Net architecture and the Unified output of enhanced images, underpinning our design philosophy.

U-FISH has been developed to address the challenges posed by significant variations in experimental conditions, hybridization targets, and imaging parameters across different data sources. These variations result in diverse image backgrounds and varying signal spot features. Conventional algorithms and parameter settings often fall short in accommodating the requirements of all these diverse image types. To overcome this limitation, we have devised a novel image enhancement approach based on the U-Net model, aimed at achieving a standardized output format for images.

Key points about U-FISH:

1. Diverse dataset: 4000+ images with approximately 1.6 million targets from seven sources, including 2 simulated datasets and 5 real datasets.
2. Small model: Achieve state-of-the-art performace with only 160k parameters (ONNX file size: 680kB).
3. 3D support: Support detection FIHS spots in 3D images.
4. Scalability: Support large-scale data storage formats: OME-Zarr and N5.
5. User-friendly interface: API, CLI, Napari plugin, and web application.

Installation

pip install ufish

For inference using GPU, you need to install onnxruntime-gpu:

pip install onnxruntime-gpu

GPU support

For training using GPU, you need to install PyTorch with CUDA support, see PyTorch official website for details.

Windows and AMD GPU

If you are using Windows and AMD GPU, you need to install:

pip install torch-directml

Usage

Fine-tuning example in colab:

CLI usage

# list all sub-commands:
$ python -m ufish

# using --help to see details of each sub-command, e.g.:
$ ufish predict --help

# predict one image
$ ufish predict input.tiff output.csv

# predict all images in a directory
$ ufish predict-imgs input_dir output_dir

# load a trained model and predict
$ ufish load-weights path/to/weights - predict-imgs input.tiff output.csv

# training from scratch
$ ufish train path/to/train_dir path/to/val_dir --model_save_dir path/to/save/model

# training from a pre-trained model (fine-tuning)
$ ufish load-weights path/to/weights - train path/to/train_dir path/to/val_dir --model_save_dir path/to/save/model

API usage

API for inference and evaluation:

from skimage import io
from ufish.api import UFish

ufish = UFish()

# loading model weights
ufish.load_weights()
# or from a file
# ufish.load_weights("path/to/weights")
# or download from huggingface:
# ufish.load_weights_from_internet()

# inference
img = io.imread("path/to/image")
pred_spots, enh_img = ufish.predict(img)

# plot prediction result
fig_spots = ufish.plot_result(img, pred_spots)

# evaluate
true_spots = pd.read_csv("path/to/true_spots.csv")
metrics = ufish.evaluate_result(spots, true_spots)

# plot evaluation result
fig_eval = ufish.plot_evaluate(
    img, pred_spots, true_spots, cutoff=3.0)

API for training:

from ufish.api import UFish

ufish = UFish()

# loading a pre-trained model
# if you want to train from scratch, just skip this step
ufish.load_weights("path/to/weights")

ufish.train(
    'path/to/train_dir',
    'path/to/val_dir',
    num_epochs=100,
    lr=1e-4,
    model_save_path='path/to/save/model',
)

Dataset

The dataset is available at HuggingFace Datasets:

# Make sure you have git-lfs installed (https://git-lfs.com)
git lfs install
git clone https://huggingface.co/datasets/GangCaoLab/FISH_spots

Citation

Xu W, Cai H, Zhang Q, et al. U-FISH: a fluorescent spot detector for imaging-based spatial-omics analysis and AI-assisted FISH diagnosis.[J]. Genome Biology, 2024: 2025.09.

BibTeX

@article{xu2025u,
  title={U-FISH: a fluorescent spot detector for imaging-based spatial-omics analysis and AI-assisted FISH diagnosis},
  author={Xu, Weize and Cai, Huaiyuan and Zhang, Qian and Wang, Zhengze and Yang, Jiajun and Wu, Xiaofeng and Li, Chengwen and Cui, Chenghua and Liu, Changzhi and He, Jin and others},
  journal={Genome Biology},
  volume={26},
  number={1},
  pages={261},
  year={2025},
  publisher={Springer}
}