Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept)
Abstract Background There is a growing interest in the scientific community to use computer-based software programs for the quantification of cells during physiological and pathophysiological processes. Drawbacks of computer-based methods currently used to quantify immunohistochemical staining are t...
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| Format: | Article |
| Language: | English |
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BMC
2020-07-01
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| Series: | BMC Veterinary Research |
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| Online Access: | http://link.springer.com/article/10.1186/s12917-020-02444-x |
| _version_ | 1818541893997297664 |
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| author | Valérie Dubuc Sheila Laverty Hélène Richard Monique Doré Christine Theoret |
| author_facet | Valérie Dubuc Sheila Laverty Hélène Richard Monique Doré Christine Theoret |
| author_sort | Valérie Dubuc |
| collection | DOAJ |
| description | Abstract Background There is a growing interest in the scientific community to use computer-based software programs for the quantification of cells during physiological and pathophysiological processes. Drawbacks of computer-based methods currently used to quantify immunohistochemical staining are the complexity of use, expense of software and overly-simplified descriptions of protocol thereby limiting reproducibility. The precise role of mast cells in equine cutaneous wound healing is unknown. Given the contribution of mast cells to the chronic inflammation observed in human keloid, a pathology similar to exuberant granulation tissue (EGT) in horses, mast cells might be present in high numbers in equine limb wounds predisposed to EGT. The main goal of this study was to develop a reliable and reproducible quantification method for immunostained tissues using a computer software that is widely available, at no cost, to the scientific community. A secondary goal was to conduct a proof of concept using the newly-established method to quantify mast cells during wound healing at different anatomical sites (body and limb) in horses to see if a different pattern is observed in limb wounds, which are predisposed to EGT. Results A good intraclass correlation coefficient (ICC, 0.67 p < 0.05) was found between the computer-based ImageJ method and manual counting. An excellent intra-operator ICC of 0.90 (p < 0.01) was found for the ImageJ quantification method while a good interoperator ICC of 0.69 (p < 0.01) was measured. No significant difference was observed between the variation of the ImageJ and that of the manual counting method. Mast cells were localized below the epidermis, around cutaneous appendages and blood vessels. Mast cell numbers did not differ significantly in relation to anatomical location or time of healing. Conclusions The computer-based quantification method developed is reliable, reproducible, available, cost-free and could be used to study different physiological and pathological processes using immunohistochemistry. |
| first_indexed | 2024-12-11T22:15:06Z |
| format | Article |
| id | doaj.art-0589ac26894b413088fe5b767e37255d |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-12-11T22:15:06Z |
| publishDate | 2020-07-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-0589ac26894b413088fe5b767e37255d2022-12-22T00:48:37ZengBMCBMC Veterinary Research1746-61482020-07-011611810.1186/s12917-020-02444-xDevelopment of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept)Valérie Dubuc0Sheila Laverty1Hélène Richard2Monique Doré3Christine Theoret4Department of Veterinary Biomedicine, Faculté de Médecine Vétérinaire, Université de MontréalDepartment of Clinical Sciences, Faculté de Médecine Vétérinaire, Université de MontréalDepartment of Clinical Sciences, Faculté de Médecine Vétérinaire, Université de MontréalDepartment of Pathology and Microbiology, Faculté de Médecine Vétérinaire, Université de MontréalDepartment of Veterinary Biomedicine, Faculté de Médecine Vétérinaire, Université de MontréalAbstract Background There is a growing interest in the scientific community to use computer-based software programs for the quantification of cells during physiological and pathophysiological processes. Drawbacks of computer-based methods currently used to quantify immunohistochemical staining are the complexity of use, expense of software and overly-simplified descriptions of protocol thereby limiting reproducibility. The precise role of mast cells in equine cutaneous wound healing is unknown. Given the contribution of mast cells to the chronic inflammation observed in human keloid, a pathology similar to exuberant granulation tissue (EGT) in horses, mast cells might be present in high numbers in equine limb wounds predisposed to EGT. The main goal of this study was to develop a reliable and reproducible quantification method for immunostained tissues using a computer software that is widely available, at no cost, to the scientific community. A secondary goal was to conduct a proof of concept using the newly-established method to quantify mast cells during wound healing at different anatomical sites (body and limb) in horses to see if a different pattern is observed in limb wounds, which are predisposed to EGT. Results A good intraclass correlation coefficient (ICC, 0.67 p < 0.05) was found between the computer-based ImageJ method and manual counting. An excellent intra-operator ICC of 0.90 (p < 0.01) was found for the ImageJ quantification method while a good interoperator ICC of 0.69 (p < 0.01) was measured. No significant difference was observed between the variation of the ImageJ and that of the manual counting method. Mast cells were localized below the epidermis, around cutaneous appendages and blood vessels. Mast cell numbers did not differ significantly in relation to anatomical location or time of healing. Conclusions The computer-based quantification method developed is reliable, reproducible, available, cost-free and could be used to study different physiological and pathological processes using immunohistochemistry.http://link.springer.com/article/10.1186/s12917-020-02444-xImmunohistochemistryQuantificationImageJHorseSkinWound healing |
| spellingShingle | Valérie Dubuc Sheila Laverty Hélène Richard Monique Doré Christine Theoret Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept) BMC Veterinary Research Immunohistochemistry Quantification ImageJ Horse Skin Wound healing |
| title | Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept) |
| title_full | Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept) |
| title_fullStr | Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept) |
| title_full_unstemmed | Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept) |
| title_short | Development of a computer-based quantification method for immunohistochemically-stained tissues and its application to study mast cells in equine wound healing (proof of concept) |
| title_sort | development of a computer based quantification method for immunohistochemically stained tissues and its application to study mast cells in equine wound healing proof of concept |
| topic | Immunohistochemistry Quantification ImageJ Horse Skin Wound healing |
| url | http://link.springer.com/article/10.1186/s12917-020-02444-x |
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