Deep Learning-Based Quantification of Pulmonary Hemosiderophages in Cytology Slides.
Abstract: Exercise-induced pulmonary hemorrhage (EIPH) is a common condition in sport horses with negative impact on performance. Cytology of bronchoalveolar lavage fluid by use of a scoring system is considered the most sensitive diagnostic method. Macrophages are classified depending on the degree of cytoplasmic hemosiderin content. The current gold standard is manual grading, which is however monotonous and time-consuming. We evaluated state-of-the-art deep learning-based methods for single cell macrophage classification and compared them against the performance of nine cytology experts and evaluated inter- and intra-observer variability. Additionally, we evaluated object detection methods on a novel data set of 17 completely annotated cytology whole slide images (WSI) containing 78,047 hemosiderophages. Our deep learning-based approach reached a concordance of 0.85, partially exceeding human expert concordance (0.68 to 0.86, mean of 0.73, SD of 0.04). Intra-observer variability was high (0.68 to 0.88) and inter-observer concordance was moderate (Fleiss' kappa = 0.67). Our object detection approach has a mean average precision of 0.66 over the five classes from the whole slide gigapixel image and a computation time of below two minutes. To mitigate the high inter- and intra-rater variability, we propose our automated object detection pipeline, enabling accurate, reproducible and quick EIPH scoring in WSI.
Publication Date: 2020-08-03 PubMed ID: 32747665PubMed Central: PMC7398908DOI: 10.1038/s41598-020-65958-2Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This study reports the use of deep learning methods to automatically identify and classify cells related to Exercise-induced Pulmonary Hemorrhage in horses, a task typically done manually – a labor-intensive process with notable observer variability. The automated method was found to be quick, accurate, and consistent, demonstrating potential to improve the current diagnostic methodology.
Research Context and Motivation
- The study focuses on Exercise-induced pulmonary hemorrhage (EIPH), a common issue in athletic horses that negatively influences their performance.
- The most sensitive method for diagnosing EIPH is the examination of bronchoalveolar lavage fluid (BALF) under a microscope, notably examining macrophages, a type of white blood cell.
- Macrophage classification is based on the degree of hemosiderin – a byproduct of blood degradation – present within the cells. This manual grading method is tiresome and monotonous.
- The researchers aimed to improve this process via modern deep learning techniques, comparing the automated method’s performance to that of nine human experts.
Methodology
- The team tested advanced deep learning techniques, specifically for classifying single cell macrophages.
- They compared these automated techniques against nine cytology experts to assess their effectiveness.
- Object detection methods were also evaluated on a distinct dataset of 17 fully annotated cytology whole slide images (WSI) containing 78,047 hemosiderophages (i.e., macrophages containing hemosiderin).
Findings
- The deep learning approach achieved a concordance (agreement) rate of 0.85, outperforming the average human expert concordance rate of 0.73.
- Intra-observer variability (differences in classification made by the same expert) ranged from 0.68 to 0.88, indicating significant variability in human observation.
- The object detection approach used on WSI had a mean average precision of 0.66 and could complete its computations in less than two minutes.
Conclusion
- The automated object detection pipeline proposed by the researchers has shown potential to serve as an efficient, reliable, and rapid tool for EIPH diagnosis in WSI.
- By mitigating high inter- and intra-observer variabilities, it may substantially enhance and expedite the current approach to diagnosing EIPH in horses.
Cite This Article
APA
Marzahl C, Aubreville M, Bertram CA, Stayt J, Jasensky AK, Bartenschlager F, Fragoso-Garcia M, Barton AK, Elsemann S, Jabari S, Krauth J, Madhu P, Voigt J, Hill J, Klopfleisch R, Maier A.
(2020).
Deep Learning-Based Quantification of Pulmonary Hemosiderophages in Cytology Slides.
Sci Rep, 10(1), 9795.
https://doi.org/10.1038/s41598-020-65958-2 Publication
Researcher Affiliations
- Pattern Recognition Lab, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany. c.marzahl@euroimmun.de.
- Research and Development, EUROIMMUN Medizinische Labordiagnostika AG, Lübeck, Germany. c.marzahl@euroimmun.de.
- Pattern Recognition Lab, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
- Institute of Veterinary Pathology, Freie Universität Berlin, Berlin, Germany.
- VetPath Laboratory Services, Ascot, Western, Australia.
- Laboklin GmbH und Co. KG, Bad Kissingen, Germany.
- Institute of Veterinary Pathology, Freie Universität Berlin, Berlin, Germany.
- Institute of Veterinary Pathology, Freie Universität Berlin, Berlin, Germany.
- Equine Clinic, Freie Universität Berlin, Berlin, Germany.
- Department of Neurosurgery, Universitätsklinikum Erlangen, Erlangen, Germany.
- Institute of Neuropathology, Friedrich Alexander University Erlangen-Nürnberg, Erlangen, Germany.
- Research and Development, EUROIMMUN Medizinische Labordiagnostika AG, Lübeck, Germany.
- Pattern Recognition Lab, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
- Research and Development, EUROIMMUN Medizinische Labordiagnostika AG, Lübeck, Germany.
- VetPath Laboratory Services, Ascot, Western, Australia.
- Institute of Veterinary Pathology, Freie Universität Berlin, Berlin, Germany.
- Pattern Recognition Lab, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
MeSH Terms
- Animals
- Cytological Techniques
- Deep Learning
- Hemorrhage / pathology
- Horses
- Lung Diseases / pathology
- Single-Cell Analysis
Conflict of Interest Statement
The authors declare no competing interests.
References
This article includes 41 references
- Ahmad KA, Bennett MM, Ahmad SF, Clark RH, Tolia VN. Morbidity and mortality with early pulmonary haemorrhage in preterm neonates.. Arch Dis Child Fetal Neonatal Ed 2019 Jan;104(1):F63-F68.
- Maldonado F, Parambil JG, Yi ES, Decker PA, Ryu JH. Haemosiderin-laden macrophages in the bronchoalveolar lavage fluid of patients with diffuse alveolar damage.. Eur Respir J 2009 Jun;33(6):1361-6.
- van Houten J, Long W, Mullett M, Finer N, Derleth D, McMurray B, Peliowski A, Walker D, Wold D, Sankaran K. Pulmonary hemorrhage in premature infants after treatment with synthetic surfactant: an autopsy evaluation. The American Exosurf Neonatal Study Group I, and the Canadian Exosurf Neonatal Study Group.. J Pediatr 1992 Feb;120(2 Pt 2):S40-4.
- Golde DW, Drew WL, Klein HZ, Finley TN, Cline MJ. Occult pulmonary haemorrhage in leukaemia.. Br Med J 1975 Apr 26;2(5964):166-8.
- Martínez-Martínez MU, Oostdam DAH, Abud-Mendoza C. Diffuse Alveolar Hemorrhage in Autoimmune Diseases.. Curr Rheumatol Rep 2017 May;19(5):27.
- Kahn FW, Jones JM, England DM. Diagnosis of pulmonary hemorrhage in the immunocompromised host.. Am Rev Respir Dis 1987 Jul;136(1):155-60.
- Hopkins SR, Schoene RB, Henderson WR, Spragg RG, Martin TR, West JB. Intense exercise impairs the integrity of the pulmonary blood-gas barrier in elite athletes.. Am J Respir Crit Care Med 1997 Mar;155(3):1090-4.
- Epp T. Evidence supporting exercise-induced pulmonary haemorrhage in racing greyhounds. Comp. Exerc. Physiol. 2008;5:21–32.
- Morley PS, Bromberek JL, Saulez MN, Hinchcliff KW, Guthrie AJ. Exercise-induced pulmonary haemorrhage impairs racing performance in Thoroughbred racehorses.. Equine Vet J 2015 May;47(3):358-65.
- Hinchcliff KW, Jackson MA, Morley PS, Brown JA, Dredge AE, O'Callaghan PA, McCaffrey JP, Slocombe RE, Clarke AE. Association between exercise-induced pulmonary hemorrhage and performance in Thoroughbred racehorses.. J Am Vet Med Assoc 2005 Sep 1;227(5):768-74.
- Birks EK, Durando MM, McBride S. Exercise-induced pulmonary hemorrhage.. Vet Clin North Am Equine Pract 2003 Apr;19(1):87-100.
- Hinchcliff KW, Couetil LL, Knight PK, Morley PS, Robinson NE, Sweeney CR, van Erck E. Exercise induced pulmonary hemorrhage in horses: American College of Veterinary Internal Medicine consensus statement.. J Vet Intern Med 2015 May-Jun;29(3):743-58.
- Hoffman AM. Bronchoalveolar lavage: sampling technique and guidelines for cytologic preparation and interpretation.. Vet Clin North Am Equine Pract 2008 Aug;24(2):423-35, vii-viii.
- Depecker M, Couroucé-Malblanc A, Leleu C, Genneviève V, Pitel PH, Richard EA. Comparison of two cytological methods for detecting pulmonary haemorrhage in horses.. Vet Rec 2015 Sep 26;177(12):305.
- Denk H, Künzele H, Plenk H, Rüschoff J, Seller W. Romeis mikroskopische technik. Urban und Schwarzenberg, München-Wien. Baltimore 439–450 (1989).
- Doucet MY, Viel L. Alveolar macrophage graded hemosiderin score from bronchoalveolar lavage in horses with exercise-induced pulmonary hemorrhage and controls.. J Vet Intern Med 2002 May-Jun;16(3):281-6.
- Waithe D. Object detection networks and augmented reality for cellular detection in fluorescence microscopy acquisition and analysis. bioRxiv 544833 (2019).
- Baykal E, Dogan H, Ercin ME, Ersoz S, Ekinci M. Modern convolutional object detectors for nuclei detection on pleural effusion cytology images. Multimedia Tools and Applications 1–20 (2019).
- Aubreville M, Bertram C, Klopfleisch R, Maier A. Field Of Interest Proposal for Augmented Mitotic Cell Count: A Comparison of Two Networks. Proceedings of the 12th International Joint Conference on Biomedical Engineering Systems and Technologies - Volume 2: BIOIMAGING 30–37, 10.5220/0007365700300037 (2019).
- Lowe DG. Object recognition from local scale-invariant features. ICCV 1999;99:1150–1157.
- Ojala T, Pietikäinen M, Harwood D. A comparative study of texture measures with classification based on featured distributions. PATTERN RECOGN. 1996;29:51–59.
- Dalal N, Triggs B. Histograms of oriented gradients for human detection. CVPR vol. 1, 886–893 (IEEE Computer Society, 2005).
- Maier A, Syben C, Lasser T, Riess C. A gentle introduction to deep learning in medical image processing.. Z Med Phys 2019 May;29(2):86-101.
- Ren S, He K, Girshick R, Sun J. Faster r-cnn: Towards real-time object detection with region proposal networks. Adv Neural Inf Process Syst 91–99 (2015).
- Liu W. Ssd: Single shot multibox detector. ECCV 21–37 (Springer, 2016).
- Lin TY, Goyal P, Girshick R, He K, Dollar P. Focal Loss for Dense Object Detection.. IEEE Trans Pattern Anal Mach Intell 2020 Feb;42(2):318-327.
- Everingham M, Van Gool L, Williams CK, Winn J, Zisserman A. The pascal visual object classes (voc) challenge. International journal of computer vision 2010;88:303–338.
- Lin T-Y. Microsoft coco: Common objects in context. ECCV 740–755 (Springer, 2014).
- Zou Z, Shi Z, Guo Y, Ye J. Object detection in 20 years: A survey. arXiv preprint arXiv:1905.05055 (2019).
- Mundhenk TN, Konjevod G, Sakla WA, Boakye K. A large contextual dataset for classification, detection and counting of cars with deep learning. ICCV 785–800 (Springer, 2016).
- Litjens G, Kooi T, Bejnordi BE, Setio AAA, Ciompi F, Ghafoorian M, van der Laak JAWM, van Ginneken B, Sánchez CI. A survey on deep learning in medical image analysis.. Med Image Anal 2017 Dec;42:60-88.
- Ronneberger O, Fischer P, Brox T. U-net: Convolutional networks for biomedical image segmentation. Med Image Comput Comput Assist Interv 234–241 (Springer, 2015).
- Ferlaino M. Towards deep cellular phenotyping in placental histology. arXiv preprint arXiv:1804.03270 (2018).
- Redmon J, Divvala S, Girshick R, Farhadi A. You only look once: Unified, real-time object detection. Proc IEEE Comput Soc Conf Comput Vis Pattern Recognit 779–788 (2016).
- Huang J. Speed/accuracy trade-offs for modern convolutional object detectors. Proc IEEE Comput Soc Conf Comput Vis Pattern Recognit 7310–7311 (2017).
- Aubreville M, Bertram C, Klopfleisch R, Maier A. Sliderunner. Bildverarbeitung für die Medizin 2018 309–314 (Springer, 2018).
- He K, Zhang X, Ren S, Sun J. Deep Residual Learning for Image Recognition. CVPR 770–778 (IEEE, 2016).
- Russakovsky O. Imagenet large scale visual recognition challenge. Int J Comput Vis 2015;115:211–252.
- Lin T-Y. Feature pyramid networks for object detection. Proc IEEE Comput Soc Conf Comput Vis Pattern Recognit 2117–2125 (2017).
- Paszke A. Automatic differentiation in PyTorch. NIPS Autodiff Workshop (2017).
- Bertram CA, Aubreville M, Gurtner C, Bartel A, Corner SM, Dettwiler M, Kershaw O, Noland EL, Schmidt A, Sledge DG, Smedley RC, Thaiwong T, Kiupel M, Maier A, Klopfleisch R. Computerized Calculation of Mitotic Count Distribution in Canine Cutaneous Mast Cell Tumor Sections: Mitotic Count Is Area Dependent.. Vet Pathol 2020 Mar;57(2):214-226.
Citations
This article has been cited 20 times.- Aubreville M, Wilm F, Stathonikos N, Breininger K, Donovan TA, Jabari S, Veta M, Ganz J, Ammeling J, van Diest PJ, Klopfleisch R, Bertram CA. A comprehensive multi-domain dataset for mitotic figure detection. Sci Data 2023 Jul 25;10(1):484.
- Wilm F, Ihling C, Méhes G, Terracciano L, Puget C, Klopfleisch R, Schüffler P, Aubreville M, Maier A, Mrowiec T, Breininger K. Pan-tumor T-lymphocyte detection using deep neural networks: Recommendations for transfer learning in immunohistochemistry. J Pathol Inform 2023;14:100301.
- Mahalingam-Dhingra A, Bedenice D, Mazan MR. Bronchoalveolar lavage hemosiderosis in lightly active or sedentary horses. J Vet Intern Med 2023 May-Jun;37(3):1243-1249.
- Rho J, Shin SM, Jhang K, Lee G, Song KH, Shin H, Na K, Kwon HJ, Son HY. Deep learning-based diagnosis of feline hypertrophic cardiomyopathy. PLoS One 2023;18(2):e0280438.
- Bertram CA, Marzahl C, Bartel A, Stayt J, Bonsembiante F, Beeler-Marfisi J, Barton AK, Brocca G, Gelain ME, Gläsel A, Preez KD, Weiler K, Weissenbacher-Lang C, Breininger K, Aubreville M, Maier A, Klopfleisch R, Hill J. Cytologic scoring of equine exercise-induced pulmonary hemorrhage: Performance of human experts and a deep learning-based algorithm. Vet Pathol 2023 Jan;60(1):75-85.
- Marzahl C, Hill J, Stayt J, Bienzle D, Welker L, Wilm F, Voigt J, Aubreville M, Maier A, Klopfleisch R, Breininger K, Bertram CA. Inter-species cell detection - datasets on pulmonary hemosiderophages in equine, human and feline specimens. Sci Data 2022 Jun 3;9(1):269.
- Sadeghi H, Braun HS, Panti B, Opsomer G, Bogado Pascottini O. Validation of a deep learning-based image analysis system to diagnose subclinical endometritis in dairy cows. PLoS One 2022;17(1):e0263409.
- Bertram CA, Aubreville M, Donovan TA, Bartel A, Wilm F, Marzahl C, Assenmacher CA, Becker K, Bennett M, Corner S, Cossic B, Denk D, Dettwiler M, Gonzalez BG, Gurtner C, Haverkamp AK, Heier A, Lehmbecker A, Merz S, Noland EL, Plog S, Schmidt A, Sebastian F, Sledge DG, Smedley RC, Tecilla M, Thaiwong T, Fuchs-Baumgartinger A, Meuten DJ, Breininger K, Kiupel M, Maier A, Klopfleisch R. Computer-assisted mitotic count using a deep learning-based algorithm improves interobserver reproducibility and accuracy. Vet Pathol 2022 Mar;59(2):211-226.
- Bertram CA, Stathonikos N, Donovan TA, Bartel A, Fuchs-Baumgartinger A, Lipnik K, van Diest PJ, Bonsembiante F, Klopfleisch R. Validation of digital microscopy: Review of validation methods and sources of bias. Vet Pathol 2022 Jan;59(1):26-38.
- Kittichai V, Kaewthamasorn M, Thanee S, Jomtarak R, Klanboot K, Naing KM, Tongloy T, Chuwongin S, Boonsang S. Classification for avian malaria parasite Plasmodium gallinaceum blood stages by using deep convolutional neural networks. Sci Rep 2021 Aug 19;11(1):16919.
- Aubreville M, Bertram CA, Donovan TA, Marzahl C, Maier A, Klopfleisch R. A completely annotated whole slide image dataset of canine breast cancer to aid human breast cancer research. Sci Data 2020 Nov 27;7(1):417.
- Neal SV, Rudmann DG, Corps KN. Artificial Intelligence in Veterinary Clinical Pathology-An Introduction and Review. Vet Clin Pathol 2025 Dec;54 Suppl 2(Suppl 2):S13-S29.
- Williams MG, Faber ZJ, Kelley TJ. Comparison of artificial intelligence image processing with manual leucocyte differential to score immune cell infiltration in a mouse infection model of cystic fibrosis. J Pathol Inform 2025 Apr;17:100438.
- Albuquerque C, Henriques R, Castelli M. Deep learning-based object detection algorithms in medical imaging: Systematic review. Heliyon 2025 Jan 15;11(1):e41137.
- Bauer TW, Hanna MG, Smith KD, Sirintrapun SJ, Hameed MR, Reddi D, Chang BS, Ardon O, Zhou X, Lewis JV, Dayal S, Chiweshe J, Ferber D, Sutcu AE, White M. A multicenter study to evaluate the analytical precision by pathologists using the Aperio GT 450 DX. J Pathol Inform 2024 Dec;15:100401.
- Rumpf S, Zufall N, Rumpf F, Gschwendtner A. A Performance Comparison of Different YOLOv7 Networks for High-Accuracy Cell Classification in Bronchoalveolar Lavage Fluid Utilising the Adam Optimiser and Label Smoothing. J Imaging Inform Med 2025 Aug;38(4):2367-2380.
- Kittichai V, Sompong W, Kaewthamasorn M, Sasisaowapak T, Naing KM, Tongloy T, Chuwongin S, Thanee S, Boonsang S. A novel approach for identification of zoonotic trypanosome utilizing deep metric learning and vector database-based image retrieval system. Heliyon 2024 May 15;10(9):e30643.
- Lapsina S, Riond B, Hofmann-Lehmann R, Stirn M. Comparison of Sysmex XN-V body fluid mode and deep-learning-based quantification with manual techniques for total nucleated cell count and differential count for equine bronchoalveolar lavage samples. BMC Vet Res 2024 Feb 5;20(1):48.
- Fragoso-Garcia M, Wilm F, Bertram CA, Merz S, Schmidt A, Donovan T, Fuchs-Baumgartinger A, Bartel A, Marzahl C, Diehl L, Puget C, Maier A, Aubreville M, Breininger K, Klopfleisch R. Automated diagnosis of 7 canine skin tumors using machine learning on H&E-stained whole slide images. Vet Pathol 2023 Nov;60(6):865-875.
- Marzahl C, Aubreville M, Bertram CA, Maier J, Bergler C, Kröger C, Voigt J, Breininger K, Klopfleisch R, Maier A. EXACT: a collaboration toolset for algorithm-aided annotation of images with annotation version control. Sci Rep 2021 Feb 23;11(1):4343.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists