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Using DeepLabCut for 3D markerless pose estimation across species and behaviors


Noninvasive behavioral tracking of animals during experiments is critical to many scientific pursuits. Extracting the poses of animals without using markers is often essential to measuring behavioral effects in biomechanics, genetics, ethology, and neuroscience. However, extracting detailed poses without markers in dynamically changing backgrounds has been challenging. We recently introduced an open-source toolbox called DeepLabCut that builds on a state-of-the-art human pose-estimation algorithm to allow a user to train a deep neural network with limited training data to precisely track user-defined features that match human labeling accuracy. Here, we provide an updated toolbox, developed as a Python package, that includes new features such as graphical user interfaces (GUIs), performance improvements, and active-learning-based network refinement. We provide a step-by-step procedure for using DeepLabCut that guides the user in creating a tailored, reusable analysis pipeline with a graphical processing unit (GPU) in 1–12 h (depending on frame size). Additionally, we provide Docker environments and Jupyter Notebooks that can be run on cloud resources such as Google Colaboratory.

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Fig. 1: Pose estimation with DeepLabCut.
Fig. 2: DeepLabCut workflow.
Fig. 3: Methods for frame selection.
Fig. 4: Labeling GUI.
Fig. 5: Evaluation of results.
Fig. 6: Refinement tools.
Fig. 7: 3D pose estimation of a cheetah.

Data and code availability

The code is fully available at Other inquiries should be made to the corresponding author (M.W.M.).


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DeepLabCut is an open-source tool on GitHub and has benefited from suggestions and edits by many individuals, including R. Eichler, J. Rauber, R. Warren, T. Abe, H. Wu, and J. Saunders. In particular, the authors thank R. Eichler for input on the modularized version. The authors thank the members of the Bethge Lab for providing the initial version of the Docker container. We also thank M. Li, J. Li, and D. Robson for use of the zebrafish image; B. Rogers for use of the horse images; and K. Cury for the fly images. The authors are grateful to E. Insafutdinov and C. Lassner for suggestions on how to best use the TensorFlow implementation of DeeperCut. We also thank A. Hoffmann, P. Mamidanna, and G. Kane for comments throughout this project. Last, the authors thank the Ann van Dyk Cheetah Centre (Pretoria, South Africa) for kindly providing access to their cheetahs. The authors thank NVIDIA Corporation for GPU grants to both M.W.M. and A.M. A.M. acknowledges a Marie Sklodowska-Curie International Fellowship within the 7th European Community Framework Program under grant agreement no. 622943. A.P. acknowledges an Oppenheimer Memorial Trust Fellowship and the National Research Foundation of South Africa (grant 99380). M.B. acknowledges funding from the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) via the Collaborative Research Center (Projektnummer 276693517–SFB 1233: Robust Vision) and by the German Federal Ministry of Education and Research through the Tübingen AI Center (FKZ 01IS18039A). M.W.M. acknowledges a Rowland Fellowship from the Rowland Institute at Harvard.

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Authors and Affiliations



Conceptualization: A.M., T.N., and M.W.M. A.M., T.N., and M.W.M. wrote the code. A.P. provided the cheetah data; A.C.C. labeled the cheetah data; A.C.C., A.M., and A.P. analyzed the cheetah data. M.W.M., A.M., and T.N. wrote the manuscript with input from all authors. M.W.M. and M.B. supervised the project.

Corresponding author

Correspondence to Mackenzie Weygandt Mathis.

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The authors declare no competing interests.

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Peer review information: Nature Protocols thanks Gonzalo G. de Polavieja and other anonymous reviewer(s) for their contribution to the peer review of this work.

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Mathis, A. et al. Nat. Neurosci. 21, 1281–1289 (2018):

Supplementary information

Supplementary Video 1

A video of the sequence described in Fig. 7c. While the images created in Fig. 7c included 256 labeled images (99% training set split, from different sessions, cameras, and perspectives), the video was created after additional training with data from different videos (908 images, 95% training set split). This network was also used for Fig. 7e.

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Nath, T., Mathis, A., Chen, A.C. et al. Using DeepLabCut for 3D markerless pose estimation across species and behaviors. Nat Protoc 14, 2152–2176 (2019).

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