Cefa Karabağ
Current Organizations / Institutes
The organizations/institutes from the scholar's latest article
Previous Organizations / Institutes
The organizations / institutes from the scholar's other articles in the past ten years
Research Subjects
Cefa Karabağ has a total of 30 co-authors. Here are the top co-authors by number of shared publications:
- Constantino Carlos Reyes-Aldasoro
City St. George’s, University of London
19 shared publications - Martin L. Jones
Electron Microscopy Science Technology Platform Francis Crick Institute London UK
7 shared publications Christopher J. Peddie
Affiliation Electron Microscopy Science Technology Platform, The Francis Crick Institute, London, United Kingdom
4 shared publications- Lucy M. Collinson
Electron Microscopy Science Technology Platform, The Francis Crick Institute, London, NW1 1AT, UK
4 shared publications Anne E. Weston
Affiliation Electron Microscopy Science Technology Platform, The Francis Crick Institute, London, United Kingdom
3 shared publications- Carlos Brito-Loeza
Universidad Autonoma de Yucatan
3 shared publications
Latest Publications
- Open Access
- 12 November 2025
- Published by MDPI AG in Journal of Imaging
- Vol. 11 (11) , 409
Image Matching for UAV Geolocation: Classical and Deep Learning Approaches
Today, unmanned aerial vehicles (UAVs) are heavily dependent on Global Navigation Satellite Systems (GNSSs) for positioning and navigation. However, GNSS signals are vulnerable to jamming and spoofing attacks. This poses serious security risks, especially for military operations and critical civilian missions. In order to solve this problem, an image-based geolocation system has been developed that eliminates GNSS dependency. The proposed system estimates the geographical location of the UAV by matching the aerial images taken by the UAV with previously georeferenced high-resolution satellite images. For this purpose, common visual features were determined between satellite and UAV images and matching operations were carried out using methods based on the homography matrix. Thanks to image processing, a significant relationship has been established between the area where the UAV is located and the geographical coordinates, and reliable positioning is ensured even in cases where GNSS signals cannot be used. Within the scope of the study, traditional methods such as SIFT, AKAZE, and Multiple Template Matching were compared with learning-based methods including SuperPoint, SuperGlue, and LoFTR. The results showed that deep learning-based approaches can make successful matches, especially at high altitudes.
- 12 November 2025
- Published by MDPI AG in Journal of Imaging
- Vol. 11 (11) , 409
Open AccessImage Matching for UAV Geolocation: Classical and Deep Learning Approaches
Today, unmanned aerial vehicles (UAVs) are heavily dependent on Global Navigation Satellite Systems (GNSSs) for positioning and navigation. However, GNSS signals are vulnerable to jamming and spoofing attacks. This poses serious security risks, especially for military operations and critical civilian missions. In order to solve this problem, an image-based geolocation system has been developed that eliminates GNSS dependency. The proposed system estimates the geographical location of the UAV by matching the aerial images taken by the UAV with previously georeferenced high-resolution satellite images. For this purpose, common visual features were determined between satellite and UAV images and matching operations were carried out using methods based on the homography matrix. Thanks to image processing, a significant relationship has been established between the area where the UAV is located and the geographical coordinates, and reliable positioning is ensured even in cases where GNSS signals cannot be used. Within the scope of the study, traditional methods such as SIFT, AKAZE, and Multiple Template Matching were compared with learning-based methods including SuperPoint, SuperGlue, and LoFTR. The results showed that deep learning-based approaches can make successful matches, especially at high altitudes.
- Open Access
- 14 September 2025
- Published by MDPI AG in Journal of Imaging
- Vol. 11 (9) , 315
From Detection to Motion-Based Classification: A Two-Stage Approach for T. cruzi Identification in Video Sequences
Chagas disease, caused by Trypanosoma cruzi (T. cruzi), remains a significant public health challenge in Latin America. Traditional diagnostic methods relying on manual microscopy suffer from low sensitivity, subjective interpretation, and poor performance in suboptimal conditions. This study presents a novel computer vision framework integrating motion analysis with deep learning for automated T. cruzi detection in microscopic videos. Our motion-based detection pipeline leverages parasite motility as a key discriminative feature, employing frame differencing, morphological processing, and DBSCAN clustering across 23 microscopic videos. This approach effectively addresses limitations of static image analysis in challenging conditions including noisy backgrounds, uneven illumination, and low contrast. From motion-identified regions, patches were extracted for classification. MobileNetV2 achieved superior performance with 99.63% accuracy, 100% precision, 99.12% recall, and an AUC-ROC of 1.0. Additionally, YOLOv5 and YOLOv8 models (Nano, Small, Medium variants) were trained on 43 annotated videos, with YOLOv5-Nano and YOLOv8-Nano demonstrating excellent detection capability on unseen test data. This dual-stage framework offers a practical, computationally efficient solution for automated Chagas diagnosis, particularly valuable for resource-constrained laboratories with poor imaging quality.- 14 September 2025
- Published by MDPI AG in Journal of Imaging
- Vol. 11 (9) , 315
Open AccessFrom Detection to Motion-Based Classification: A Two-Stage Approach for T. cruzi Identification in Video Sequences
Chagas disease, caused by Trypanosoma cruzi (T. cruzi), remains a significant public health challenge in Latin America. Traditional diagnostic methods relying on manual microscopy suffer from low sensitivity, subjective interpretation, and poor performance in suboptimal conditions. This study presents a novel computer vision framework integrating motion analysis with deep learning for automated T. cruzi detection in microscopic videos. Our motion-based detection pipeline leverages parasite motility as a key discriminative feature, employing frame differencing, morphological processing, and DBSCAN clustering across 23 microscopic videos. This approach effectively addresses limitations of static image analysis in challenging conditions including noisy backgrounds, uneven illumination, and low contrast. From motion-identified regions, patches were extracted for classification. MobileNetV2 achieved superior performance with 99.63% accuracy, 100% precision, 99.12% recall, and an AUC-ROC of 1.0. Additionally, YOLOv5 and YOLOv8 models (Nano, Small, Medium variants) were trained on 43 annotated videos, with YOLOv5-Nano and YOLOv8-Nano demonstrating excellent detection capability on unseen test data. This dual-stage framework offers a practical, computationally efficient solution for automated Chagas diagnosis, particularly valuable for resource-constrained laboratories with poor imaging quality. - 24 July 2024
- Published by Springer Nature
- p. 202-216
Image Processing and Machine Learning Techniques for Chagas Disease Detection and Identification
- 24 July 2024
- Published by Springer Nature
- p. 202-216
Image Processing and Machine Learning Techniques for Chagas Disease Detection and Identification
- Open Access
- 13 March 2024
- Published by Institute of Electrical and Electronics Engineers (IEEE) in IEEE Access
- Vol. 12, 40412-40424
DRD-UNet, a UNet-Like Architecture for Multi-Class Breast Cancer Semantic Segmentation
Staining of histological slides with Hematoxylin and Eosin is widely used in clinical and laboratory settings as these dyes reveal nuclear structures as well as cytoplasm and collagen. For cancer diagnosis, these slides are used to recognize tissues and morphological changes. Tissue semantic segmentation is therefore important and at the same time a challenging and time-consuming task. This paper describes a UNet-like deep learning architecture called DRD-UNet , which adds a novel processing block called DRD (Dilation, Residual, and Dense block) to a UNet architecture. DRD is formed by the combination of dilated convolutions (D), residual connections (R), and dense layers (D). DRD-UNet was applied to the multi-class (tumor, stroma, inflammatory, necrosis, and other) semantic segmentation of histological images from breast cancer samples stained with Hematoxylin and Eosin. The histological images were released through the Breast Cancer Semantic Segmentation (BCSS) Challenge. DRD-UNet outperformed the original UNet architecture and 15 other UNet-based architectures on the segmentation of 12,930 image patches extracted from regions of interest that ranged in size between 1036 × 1222 to 6813 × 7360 pixels. DRD-UNet obtained the best performance as measured with Jaccard similarity index, Dice coefficient, in a per-class comparison and accuracy for overall segmentation.
- 13 March 2024
- Published by Institute of Electrical and Electronics Engineers (IEEE) in IEEE Access
- Vol. 12, 40412-40424
Open AccessDRD-UNet, a UNet-Like Architecture for Multi-Class Breast Cancer Semantic Segmentation
Staining of histological slides with Hematoxylin and Eosin is widely used in clinical and laboratory settings as these dyes reveal nuclear structures as well as cytoplasm and collagen. For cancer diagnosis, these slides are used to recognize tissues and morphological changes. Tissue semantic segmentation is therefore important and at the same time a challenging and time-consuming task. This paper describes a UNet-like deep learning architecture called DRD-UNet , which adds a novel processing block called DRD (Dilation, Residual, and Dense block) to a UNet architecture. DRD is formed by the combination of dilated convolutions (D), residual connections (R), and dense layers (D). DRD-UNet was applied to the multi-class (tumor, stroma, inflammatory, necrosis, and other) semantic segmentation of histological images from breast cancer samples stained with Hematoxylin and Eosin. The histological images were released through the Breast Cancer Semantic Segmentation (BCSS) Challenge. DRD-UNet outperformed the original UNet architecture and 15 other UNet-based architectures on the segmentation of 12,930 image patches extracted from regions of interest that ranged in size between 1036 × 1222 to 6813 × 7360 pixels. DRD-UNet obtained the best performance as measured with Jaccard similarity index, Dice coefficient, in a per-class comparison and accuracy for overall segmentation.
- Open Access
Relationship Between Irregularities of the Nuclear Envelope and Mitochondria in HeLa cells Observed with Electron Microscopy
This paper describes a methodology to analyse the complexity of HeLa cells as observed with electron microscopy, in particular the relationship between mitochondria and the roughness of the nuclear envelope as reflected by the invaginations of the surface. For this purpose, several segmentation mitochondria algorithms were quantitatively compared, namely: Topology, Image Processing, Topology and Image Processing, and Deep Learning, which provided the highest accuracy. The invaginations were successfully segmented with one image processing algorithm. Metrics were extracted for both structures and correlations between the mitochondria and invaginations were explored for 25 segmented cells. It was found that there was a positive correlation between the volume of invaginations and the volume of mitochondria, and negative correlations between the number and the mean volume of mitochondria, and between the volume of the cytoplasm and the aspect ratio of mitochondria. These results suggest that there is a relationship between the shape of a cell, its nucleus and its mitochondria; as well as a relationship between the number of mitochondria and their shapes. Whilst these results were obtained from a single cell line and a relatively small number of cells, they encourage further study as the methodology proposed can be easily applied to other cells and settings. Code and data are freely available. HeLa images are available from http://dx.doi.org/10.6019/EMPIAR-10094 , code from https://github.com/reyesaldasoro/MitoEM , and segmented nuclei, cells, invaginations and mitochondria from https://github.com/reyesaldasoro/HeLa_Cell_Data .
Open AccessRelationship Between Irregularities of the Nuclear Envelope and Mitochondria in HeLa cells Observed with Electron Microscopy
This paper describes a methodology to analyse the complexity of HeLa cells as observed with electron microscopy, in particular the relationship between mitochondria and the roughness of the nuclear envelope as reflected by the invaginations of the surface. For this purpose, several segmentation mitochondria algorithms were quantitatively compared, namely: Topology, Image Processing, Topology and Image Processing, and Deep Learning, which provided the highest accuracy. The invaginations were successfully segmented with one image processing algorithm. Metrics were extracted for both structures and correlations between the mitochondria and invaginations were explored for 25 segmented cells. It was found that there was a positive correlation between the volume of invaginations and the volume of mitochondria, and negative correlations between the number and the mean volume of mitochondria, and between the volume of the cytoplasm and the aspect ratio of mitochondria. These results suggest that there is a relationship between the shape of a cell, its nucleus and its mitochondria; as well as a relationship between the number of mitochondria and their shapes. Whilst these results were obtained from a single cell line and a relatively small number of cells, they encourage further study as the methodology proposed can be easily applied to other cells and settings. Code and data are freely available. HeLa images are available from http://dx.doi.org/10.6019/EMPIAR-10094 , code from https://github.com/reyesaldasoro/MitoEM , and segmented nuclei, cells, invaginations and mitochondria from https://github.com/reyesaldasoro/HeLa_Cell_Data .
- Open Access
- 1 March 2023
- Published by MDPI AG in Journal of Imaging
- Vol. 9 (3) , 59
Impact of Training Data, Ground Truth and Shape Variability in the Deep Learning-Based Semantic Segmentation of HeLa Cells Observed with Electron Microscopy
This paper investigates the impact of the amount of training data and the shape variability on the segmentation provided by the deep learning architecture U-Net. Further, the correctness of ground truth (GT) was also evaluated. The input data consisted of a three-dimensional set of images of HeLa cells observed with an electron microscope with dimensions . From there, a smaller region of interest (ROI) of was cropped and manually delineated to obtain the ground truth necessary for a quantitative evaluation. A qualitative evaluation was performed on the slices due to the lack of ground truth. Pairs of patches of data and labels for the classes nucleus, nuclear envelope, cell and background were generated to train U-Net architectures from scratch. Several training strategies were followed, and the results were compared against a traditional image processing algorithm. The correctness of GT, that is, the inclusion of one or more nuclei within the region of interest was also evaluated. The impact of the extent of training data was evaluated by comparing results from 36,000 pairs of data and label patches extracted from the odd slices in the central region, to 135,000 patches obtained from every other slice in the set. Then, 135,000 patches from several cells from the slices were generated automatically using the image processing algorithm. Finally, the two sets of 135,000 pairs were combined to train once more with 270,000 pairs. As would be expected, the accuracy and Jaccard similarity index improved as the number of pairs increased for the ROI. This was also observed qualitatively for the slices. When the slices were segmented with U-Nets trained with 135,000 pairs, the architecture trained with automatically generated pairs provided better results than the architecture trained with the pairs from the manually segmented ground truths. This suggests that the pairs that were extracted automatically from many cells provided a better representation of the four classes of the various cells in the slice than those pairs that were manually segmented from a single cell. Finally, the two sets of 135,000 pairs were combined, and the U-Net trained with these provided the best results.- 1 March 2023
- Published by MDPI AG in Journal of Imaging
- Vol. 9 (3) , 59
Open AccessImpact of Training Data, Ground Truth and Shape Variability in the Deep Learning-Based Semantic Segmentation of HeLa Cells Observed with Electron Microscopy
This paper investigates the impact of the amount of training data and the shape variability on the segmentation provided by the deep learning architecture U-Net. Further, the correctness of ground truth (GT) was also evaluated. The input data consisted of a three-dimensional set of images of HeLa cells observed with an electron microscope with dimensions . From there, a smaller region of interest (ROI) of was cropped and manually delineated to obtain the ground truth necessary for a quantitative evaluation. A qualitative evaluation was performed on the slices due to the lack of ground truth. Pairs of patches of data and labels for the classes nucleus, nuclear envelope, cell and background were generated to train U-Net architectures from scratch. Several training strategies were followed, and the results were compared against a traditional image processing algorithm. The correctness of GT, that is, the inclusion of one or more nuclei within the region of interest was also evaluated. The impact of the extent of training data was evaluated by comparing results from 36,000 pairs of data and label patches extracted from the odd slices in the central region, to 135,000 patches obtained from every other slice in the set. Then, 135,000 patches from several cells from the slices were generated automatically using the image processing algorithm. Finally, the two sets of 135,000 pairs were combined to train once more with 270,000 pairs. As would be expected, the accuracy and Jaccard similarity index improved as the number of pairs increased for the ROI. This was also observed qualitatively for the slices. When the slices were segmented with U-Nets trained with 135,000 pairs, the architecture trained with automatically generated pairs provided better results than the architecture trained with the pairs from the manually segmented ground truths. This suggests that the pairs that were extracted automatically from many cells provided a better representation of the four classes of the various cells in the slice than those pairs that were manually segmented from a single cell. Finally, the two sets of 135,000 pairs were combined, and the U-Net trained with these provided the best results. - Open Access
Evaluation of training data, ground truth and shape variability in the semantic segmentation of HeLa cells observed with electron microscopy
This paper describes the quantitative and qualitative evaluation of training data, ground truth and the variability of the shape of HeLa cells as observed with electron microscopy in the process of semantic segmentation. HeLa cells are a widely used immortal cell line, but the principles described here could apply to other cells. From a data set of 8, 000 × 8, 000 × 517 voxels, a region of interest (ROI) of 2, 000 × 2, 000 × 300 was cropped for quantitative evaluation with the qualitative evaluation performed on a larger section of interest. Patches of data and labels were used to train a U-Net architecture from scratch, which was compared also with a traditional image processing algorithm. The correctness of ground truth, that is, the inclusion of one or more nuclei within a region of interest was evaluated with a hand-drawn ground truth of the ROI. The impact of the extent of training data was evaluated by comparing results from 36,000 data and label patches extracted from the odd slices in the central region, to 135,000 patches obtained from every other slice in the set. Accuracy and Jaccard similarity index were higher for the larger training set with multiple nuclei in the ground truth. However, when the U-Nets were used to segment the section of interest beyond the ROI, results were not as good as expected, suggesting that the very complicated nature of the shapes of HeLa cells cannot be learned from a single cell. Data, code and ground truth have been made publicly released through Empair, GitHub and Zenodo.
Open AccessEvaluation of training data, ground truth and shape variability in the semantic segmentation of HeLa cells observed with electron microscopy
This paper describes the quantitative and qualitative evaluation of training data, ground truth and the variability of the shape of HeLa cells as observed with electron microscopy in the process of semantic segmentation. HeLa cells are a widely used immortal cell line, but the principles described here could apply to other cells. From a data set of 8, 000 × 8, 000 × 517 voxels, a region of interest (ROI) of 2, 000 × 2, 000 × 300 was cropped for quantitative evaluation with the qualitative evaluation performed on a larger section of interest. Patches of data and labels were used to train a U-Net architecture from scratch, which was compared also with a traditional image processing algorithm. The correctness of ground truth, that is, the inclusion of one or more nuclei within a region of interest was evaluated with a hand-drawn ground truth of the ROI. The impact of the extent of training data was evaluated by comparing results from 36,000 data and label patches extracted from the odd slices in the central region, to 135,000 patches obtained from every other slice in the set. Accuracy and Jaccard similarity index were higher for the larger training set with multiple nuclei in the ground truth. However, when the U-Nets were used to segment the section of interest beyond the ROI, results were not as good as expected, suggesting that the very complicated nature of the shapes of HeLa cells cannot be learned from a single cell. Data, code and ground truth have been made publicly released through Empair, GitHub and Zenodo.
- Open Access
Classification and Visualisation of Normal and Abnormal Radiographs; A Comparison between Eleven Convolutional Neural Network Architectures
This paper investigates the classification of radiographic images with eleven convolutional neural network (CNN) architectures (GoogleNet, VGG-19, AlexNet, SqueezeNet, ResNet-18, Inception-v3, ResNet-50, VGG-16, ResNet-101, DenseNet-201 and Inception-ResNet-v2). The CNNs were used to classify a series of wrist radiographs from the Stanford Musculoskeletal Radiographs (MURA) dataset into two classes—normal and abnormal. The architectures were compared for different hyper-parameters against accuracy and Cohen’s kappa coefficient. The best two results were then explored with data augmentation. Without the use of augmentation, the best results were provided by Inception-ResNet-v2 (Mean accuracy = 0.723, Mean kappa = 0.506). These were significantly improved with augmentation to Inception-ResNet-v2 (Mean accuracy = 0.857, Mean kappa = 0.703). Finally, Class Activation Mapping was applied to interpret activation of the network against the location of an anomaly in the radiographs.
Open AccessClassification and Visualisation of Normal and Abnormal Radiographs; A Comparison between Eleven Convolutional Neural Network Architectures
This paper investigates the classification of radiographic images with eleven convolutional neural network (CNN) architectures (GoogleNet, VGG-19, AlexNet, SqueezeNet, ResNet-18, Inception-v3, ResNet-50, VGG-16, ResNet-101, DenseNet-201 and Inception-ResNet-v2). The CNNs were used to classify a series of wrist radiographs from the Stanford Musculoskeletal Radiographs (MURA) dataset into two classes—normal and abnormal. The architectures were compared for different hyper-parameters against accuracy and Cohen’s kappa coefficient. The best two results were then explored with data augmentation. Without the use of augmentation, the best results were provided by Inception-ResNet-v2 (Mean accuracy = 0.723, Mean kappa = 0.506). These were significantly improved with augmentation to Inception-ResNet-v2 (Mean accuracy = 0.857, Mean kappa = 0.703). Finally, Class Activation Mapping was applied to interpret activation of the network against the location of an anomaly in the radiographs.
- Open Access
Classification and Visualisation of Normal and Abnormal Radiographs; a comparison between Eleven Convolutional Neural Network Architectures
This paper investigates the classification of radiographic images with eleven convolutional neural network (CNN) architectures ( GoogleNet, VGG-19, AlexNet, SqueezeNet, ResNet-18, Inception-v3, ResNet-50, VGG-16, ResNet-101, DenseNet-201 and Inception-ResNet-v2 ). The CNNs were used to classify a series of wrist radiographs from the Stanford Musculoskeletal Radiographs (MURA) dataset into two classes - normal and abnormal. The architectures were compared for different hyper-parameters against accuracy and Cohen’s kappa coefficient. The best two results were then explored with data augmentation. Without the use of augmentation, the best results were provided by Inception-Resnet-v2 (Mean accuracy = 0.723 , Mean kappa = 0.506 ). These were significantly improved with augmentation to Inception-Resnet-v2 (Mean accuracy = 0.857 , Mean kappa = 0.703 ). Finally, Class Activation Mapping was applied to interpret activation of the network against the location of an anomaly in the radiographs.
Open AccessClassification and Visualisation of Normal and Abnormal Radiographs; a comparison between Eleven Convolutional Neural Network Architectures
This paper investigates the classification of radiographic images with eleven convolutional neural network (CNN) architectures ( GoogleNet, VGG-19, AlexNet, SqueezeNet, ResNet-18, Inception-v3, ResNet-50, VGG-16, ResNet-101, DenseNet-201 and Inception-ResNet-v2 ). The CNNs were used to classify a series of wrist radiographs from the Stanford Musculoskeletal Radiographs (MURA) dataset into two classes - normal and abnormal. The architectures were compared for different hyper-parameters against accuracy and Cohen’s kappa coefficient. The best two results were then explored with data augmentation. Without the use of augmentation, the best results were provided by Inception-Resnet-v2 (Mean accuracy = 0.723 , Mean kappa = 0.506 ). These were significantly improved with augmentation to Inception-Resnet-v2 (Mean accuracy = 0.857 , Mean kappa = 0.703 ). Finally, Class Activation Mapping was applied to interpret activation of the network against the location of an anomaly in the radiographs.
- Open Access
- 1 June 2021
- Published by MDPI AG in Journal of Imaging
- Vol. 7 (6) , 93
Volumetric Semantic Instance Segmentation of the Plasma Membrane of HeLa Cells
In this work, an unsupervised volumetric semantic instance segmentation of the plasma membrane of HeLa cells as observed with serial block face scanning electron microscopy is described. The resin background of the images was segmented at different slices of a 3D stack of 518 slices with 8192 × 8192 pixels each. The background was used to create a distance map, which helped identify and rank the cells by their size at each slice. The centroids of the cells detected at different slices were linked to identify them as a single cell that spanned a number of slices. A subset of these cells, i.e., the largest ones and those not close to the edges were selected for further processing. The selected cells were then automatically cropped to smaller regions of interest of 2000 × 2000 × 300 voxels that were treated as cell instances. Then, for each of these volumes, the nucleus was segmented, and the cell was separated from any neighbouring cells through a series of traditional image processing steps that followed the plasma membrane. The segmentation process was repeated for all the regions of interest previously selected. For one cell for which the ground truth was available, the algorithm provided excellent results in Accuracy (AC) and the Jaccard similarity Index (JI): nucleus: JI , AC , cell including nucleus JI , AC , cell excluding nucleus JI , AC . A limitation of the algorithm for the plasma membrane segmentation was the presence of background. In samples with tightly packed cells, this may not be available. When tested for these conditions, the segmentation of the nuclear envelope was still possible. All the code and data were released openly through GitHub, Zenodo and EMPIAR.- 1 June 2021
- Published by MDPI AG in Journal of Imaging
- Vol. 7 (6) , 93
Open AccessVolumetric Semantic Instance Segmentation of the Plasma Membrane of HeLa Cells
In this work, an unsupervised volumetric semantic instance segmentation of the plasma membrane of HeLa cells as observed with serial block face scanning electron microscopy is described. The resin background of the images was segmented at different slices of a 3D stack of 518 slices with 8192 × 8192 pixels each. The background was used to create a distance map, which helped identify and rank the cells by their size at each slice. The centroids of the cells detected at different slices were linked to identify them as a single cell that spanned a number of slices. A subset of these cells, i.e., the largest ones and those not close to the edges were selected for further processing. The selected cells were then automatically cropped to smaller regions of interest of 2000 × 2000 × 300 voxels that were treated as cell instances. Then, for each of these volumes, the nucleus was segmented, and the cell was separated from any neighbouring cells through a series of traditional image processing steps that followed the plasma membrane. The segmentation process was repeated for all the regions of interest previously selected. For one cell for which the ground truth was available, the algorithm provided excellent results in Accuracy (AC) and the Jaccard similarity Index (JI): nucleus: JI , AC , cell including nucleus JI , AC , cell excluding nucleus JI , AC . A limitation of the algorithm for the plasma membrane segmentation was the presence of background. In samples with tightly packed cells, this may not be available. When tested for these conditions, the segmentation of the nuclear envelope was still possible. All the code and data were released openly through GitHub, Zenodo and EMPIAR.