OC_Finder: Osteoclast Segmentation, Counting, and Classification Using Watershed and Deep Learning

Osteoclasts are multinucleated cells that exclusively resorb bone matrix proteins and minerals on the bone surface. They differentiate from monocyte/macrophage lineage cells in the presence of osteoclastogenic cytokines such as the receptor activator of nuclear factor-κB ligand (RANKL) and are stain...

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Main Authors: Xiao Wang, Mizuho Kittaka, Yilin He, Yiwei Zhang, Yasuyoshi Ueki, Daisuke Kihara
Format: Article
Language:English
Published: Frontiers Media S.A. 2022-03-01
Series:Frontiers in Bioinformatics
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fbinf.2022.819570/full
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author Xiao Wang
Mizuho Kittaka
Mizuho Kittaka
Yilin He
Yiwei Zhang
Yasuyoshi Ueki
Yasuyoshi Ueki
Daisuke Kihara
Daisuke Kihara
Daisuke Kihara
author_facet Xiao Wang
Mizuho Kittaka
Mizuho Kittaka
Yilin He
Yiwei Zhang
Yasuyoshi Ueki
Yasuyoshi Ueki
Daisuke Kihara
Daisuke Kihara
Daisuke Kihara
author_sort Xiao Wang
collection DOAJ
description Osteoclasts are multinucleated cells that exclusively resorb bone matrix proteins and minerals on the bone surface. They differentiate from monocyte/macrophage lineage cells in the presence of osteoclastogenic cytokines such as the receptor activator of nuclear factor-κB ligand (RANKL) and are stained positive for tartrate-resistant acid phosphatase (TRAP). In vitro osteoclast formation assays are commonly used to assess the capacity of osteoclast precursor cells for differentiating into osteoclasts wherein the number of TRAP-positive multinucleated cells is counted as osteoclasts. Osteoclasts are manually identified on cell culture dishes by human eyes, which is a labor-intensive process. Moreover, the manual procedure is not objective and results in lack of reproducibility. To accelerate the process and reduce the workload for counting the number of osteoclasts, we developed OC_Finder, a fully automated system for identifying osteoclasts in microscopic images. OC_Finder consists of cell image segmentation with a watershed algorithm and cell classification using deep learning. OC_Finder detected osteoclasts differentiated from wild-type and Sh3bp2KI/+ precursor cells at a 99.4% accuracy for segmentation and at a 98.1% accuracy for classification. The number of osteoclasts classified by OC_Finder was at the same accuracy level with manual counting by a human expert. OC_Finder also showed consistent performance on additional datasets collected with different microscopes with different settings by different operators. Together, successful development of OC_Finder suggests that deep learning is a useful tool to perform prompt and accurate unbiased classification and detection of specific cell types in microscopic images.
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spelling doaj.art-c76e88be9b594650b3f8d83463694eaf2022-12-21T18:11:44ZengFrontiers Media S.A.Frontiers in Bioinformatics2673-76472022-03-01210.3389/fbinf.2022.819570819570OC_Finder: Osteoclast Segmentation, Counting, and Classification Using Watershed and Deep LearningXiao Wang0Mizuho Kittaka1Mizuho Kittaka2Yilin He3Yiwei Zhang4Yasuyoshi Ueki5Yasuyoshi Ueki6Daisuke Kihara7Daisuke Kihara8Daisuke Kihara9Department of Computer Science, Purdue University, West Lafayette, IN, United StatesDepartment of Biomedical Sciences and Comprehensive Care, Indiana University School of Dentistry, Indianapolis, IN, United StatesIndiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, United StatesSchool of Software Engineering, Shandong University, Jinan, ChinaDepartment of Computer Science, Rensselaer Polytechnic Institute, Troy, NY, United StatesDepartment of Biomedical Sciences and Comprehensive Care, Indiana University School of Dentistry, Indianapolis, IN, United StatesIndiana Center for Musculoskeletal Health, Indiana University School of Medicine, Indianapolis, IN, United StatesDepartment of Computer Science, Purdue University, West Lafayette, IN, United StatesDepartment of Biological Sciences, Purdue University, West Lafayette, IN, United StatesPurdue Cancer Research Institute, Purdue University, West Lafayette, IN, United StatesOsteoclasts are multinucleated cells that exclusively resorb bone matrix proteins and minerals on the bone surface. They differentiate from monocyte/macrophage lineage cells in the presence of osteoclastogenic cytokines such as the receptor activator of nuclear factor-κB ligand (RANKL) and are stained positive for tartrate-resistant acid phosphatase (TRAP). In vitro osteoclast formation assays are commonly used to assess the capacity of osteoclast precursor cells for differentiating into osteoclasts wherein the number of TRAP-positive multinucleated cells is counted as osteoclasts. Osteoclasts are manually identified on cell culture dishes by human eyes, which is a labor-intensive process. Moreover, the manual procedure is not objective and results in lack of reproducibility. To accelerate the process and reduce the workload for counting the number of osteoclasts, we developed OC_Finder, a fully automated system for identifying osteoclasts in microscopic images. OC_Finder consists of cell image segmentation with a watershed algorithm and cell classification using deep learning. OC_Finder detected osteoclasts differentiated from wild-type and Sh3bp2KI/+ precursor cells at a 99.4% accuracy for segmentation and at a 98.1% accuracy for classification. The number of osteoclasts classified by OC_Finder was at the same accuracy level with manual counting by a human expert. OC_Finder also showed consistent performance on additional datasets collected with different microscopes with different settings by different operators. Together, successful development of OC_Finder suggests that deep learning is a useful tool to perform prompt and accurate unbiased classification and detection of specific cell types in microscopic images.https://www.frontiersin.org/articles/10.3389/fbinf.2022.819570/fulldeep learningosteoclast segmentationosteoclast countingautomatic segmentationopen source software
spellingShingle Xiao Wang
Mizuho Kittaka
Mizuho Kittaka
Yilin He
Yiwei Zhang
Yasuyoshi Ueki
Yasuyoshi Ueki
Daisuke Kihara
Daisuke Kihara
Daisuke Kihara
OC_Finder: Osteoclast Segmentation, Counting, and Classification Using Watershed and Deep Learning
Frontiers in Bioinformatics
deep learning
osteoclast segmentation
osteoclast counting
automatic segmentation
open source software
title OC_Finder: Osteoclast Segmentation, Counting, and Classification Using Watershed and Deep Learning
title_full OC_Finder: Osteoclast Segmentation, Counting, and Classification Using Watershed and Deep Learning
title_fullStr OC_Finder: Osteoclast Segmentation, Counting, and Classification Using Watershed and Deep Learning
title_full_unstemmed OC_Finder: Osteoclast Segmentation, Counting, and Classification Using Watershed and Deep Learning
title_short OC_Finder: Osteoclast Segmentation, Counting, and Classification Using Watershed and Deep Learning
title_sort oc finder osteoclast segmentation counting and classification using watershed and deep learning
topic deep learning
osteoclast segmentation
osteoclast counting
automatic segmentation
open source software
url https://www.frontiersin.org/articles/10.3389/fbinf.2022.819570/full
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