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<article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xmlns:ali="http://www.niso.org/schemas/ali/1.0/" article-type="research-article" dtd-version="1.2" xml:lang="en"><front><journal-meta><journal-id journal-id-type="publisher-id">Morphology</journal-id><journal-title-group><journal-title xml:lang="en">Morphology</journal-title><trans-title-group xml:lang="ru"><trans-title>Морфология</trans-title></trans-title-group></journal-title-group><issn publication-format="print">1026-3543</issn><issn publication-format="electronic">2949-2556</issn><publisher><publisher-name xml:lang="en">Eco-Vector</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="publisher-id">642031</article-id><article-id pub-id-type="doi">10.17816/morph.642031</article-id><article-categories><subj-group subj-group-type="toc-heading" xml:lang="en"><subject>Original Study Articles</subject></subj-group><subj-group subj-group-type="toc-heading" xml:lang="ru"><subject>Оригинальные исследования</subject></subj-group><subj-group subj-group-type="article-type"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title xml:lang="en">Use of combined eosin and methylene green staining for the selective detection of eosinophilic granulocytes in the gastrointestinal mucosa</article-title><trans-title-group xml:lang="ru"><trans-title>Применение комбинированной окраски эозином и метиленовым зелёным для селективного выявления эозинофильных гранулоцитов в слизистой оболочке пищеварительного тракта</trans-title></trans-title-group><trans-title-group xml:lang="zh"><trans-title>结合伊红和亚甲基绿染色法在消化道黏膜中选择性显示嗜酸性粒细胞的应用</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0009-0002-2618-7670</contrib-id><contrib-id contrib-id-type="spin">8573-1606</contrib-id><name-alternatives><name xml:lang="en"><surname>Zabelnikova</surname><given-names>Aleksandra M.</given-names></name><name xml:lang="ru"><surname>Забельникова</surname><given-names>Александра Михайловна</given-names></name><name xml:lang="zh"><surname>Zabelnikova</surname><given-names>Aleksandra M.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><email>zabel.am@yandex.ru</email><xref ref-type="aff" rid="aff1"/><xref ref-type="aff" rid="aff2"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0001-5730-4265</contrib-id><contrib-id contrib-id-type="spin">7796-1107</contrib-id><name-alternatives><name xml:lang="en"><surname>Beznin</surname><given-names>Gleb V.</given-names></name><name xml:lang="ru"><surname>Безнин</surname><given-names>Глеб Владимирович</given-names></name><name xml:lang="zh"><surname>Beznin</surname><given-names>Gleb V.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Cand. Sci. (Medicine)</p></bio><bio xml:lang="ru"><p>канд. мед. наук</p></bio><bio xml:lang="zh"><p>Cand. Sci. (Medicine)</p></bio><email>beznin.gv@iemspb.ru</email><xref ref-type="aff" rid="aff3"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-1010-6768</contrib-id><contrib-id contrib-id-type="spin">5019-3940</contrib-id><name-alternatives><name xml:lang="en"><surname>Abdurasulova</surname><given-names>Irina N.</given-names></name><name xml:lang="ru"><surname>Абдурасулова</surname><given-names>Ирина Николаевна</given-names></name><name xml:lang="zh"><surname>Abdurasulova</surname><given-names>Irina N.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Cand. Sci. (Biology)</p></bio><bio xml:lang="ru"><p>канд. биол. наук</p></bio><bio xml:lang="zh"><p>Cand. Sci. (Biology)</p></bio><email>i_abdurasulova@mail.ru</email><xref ref-type="aff" rid="aff3"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0001-6113-3948</contrib-id><contrib-id contrib-id-type="spin">5725-8742</contrib-id><name-alternatives><name xml:lang="en"><surname>Kirik</surname><given-names>Olga V.</given-names></name><name xml:lang="ru"><surname>Кирик</surname><given-names>Ольга Викторовна</given-names></name><name xml:lang="zh"><surname>Kirik</surname><given-names>Olga V.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Cand. Sci. (Biology)</p></bio><bio xml:lang="ru"><p>канд. биол. наук</p></bio><bio xml:lang="zh"><p>Cand. Sci. (Biology)</p></bio><email>olga_kirik@mail.ru</email><xref ref-type="aff" rid="aff3"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="spin">5414-4122</contrib-id><name-alternatives><name xml:lang="en"><surname>Fedorova</surname><given-names>Elena A.</given-names></name><name xml:lang="ru"><surname>Фeдорова</surname><given-names>Елена Анатольевна</given-names></name><name xml:lang="zh"><surname>Fedorova</surname><given-names>Elena A.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Cand. Sci. (Biology)</p></bio><bio xml:lang="ru"><p>канд. биол. наук</p></bio><bio xml:lang="zh"><p>Cand. Sci. (Biology)</p></bio><email>el-fedorova2014@ya.ru</email><xref ref-type="aff" rid="aff3"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-2456-8165</contrib-id><contrib-id contrib-id-type="spin">3252-3029</contrib-id><name-alternatives><name xml:lang="en"><surname>Korzhevskii</surname><given-names>Dmitrii E.</given-names></name><name xml:lang="ru"><surname>Коржевский</surname><given-names>Дмитрий Эдуардович</given-names></name><name xml:lang="zh"><surname>Korzhevskii</surname><given-names>Dmitrii E.</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>MD, Dr. Sci. (Medicine), Professor</p></bio><bio xml:lang="ru"><p>д-р мед. наук, профессор</p></bio><bio xml:lang="zh"><p>MD, Dr. Sci. (Medicine), Professor</p></bio><email>dek2@yandex.ru</email><xref ref-type="aff" rid="aff3"/></contrib></contrib-group><aff-alternatives id="aff1"><aff><institution xml:lang="en">Institute of Experimental Medicine</institution></aff><aff><institution xml:lang="kk"></institution></aff><aff><institution xml:lang="pt"></institution></aff><aff><institution xml:lang="ru">Институт экспериментальной медицины</institution></aff><aff><institution xml:lang="zh">Institute of Experimental Medicine</institution></aff></aff-alternatives><aff-alternatives id="aff2"><aff><institution xml:lang="en">Saint-Petersburg State University</institution></aff><aff><institution xml:lang="ru">Санкт-Петербургский государственный университет</institution></aff><aff><institution xml:lang="zh">Saint-Petersburg State University</institution></aff></aff-alternatives><aff-alternatives id="aff3"><aff><institution xml:lang="en">Institute of Experimental Medicine</institution></aff><aff><institution xml:lang="ru">Институт экспериментальной медицины</institution></aff><aff><institution xml:lang="zh">Institute of Experimental Medicine</institution></aff></aff-alternatives><pub-date date-type="preprint" iso-8601-date="2025-02-02" publication-format="electronic"><day>02</day><month>02</month><year>2025</year></pub-date><pub-date date-type="pub" iso-8601-date="2024-12-28" publication-format="electronic"><day>28</day><month>12</month><year>2024</year></pub-date><volume>162</volume><issue>4</issue><issue-title xml:lang="en"/><issue-title xml:lang="ru"/><issue-title xml:lang="zh"/><fpage>416</fpage><lpage>424</lpage><history><date date-type="received" iso-8601-date="2024-11-22"><day>22</day><month>11</month><year>2024</year></date><date date-type="accepted" iso-8601-date="2025-01-10"><day>10</day><month>01</month><year>2025</year></date></history><permissions><copyright-statement xml:lang="en">Copyright ©; 2024, Eco-Vector</copyright-statement><copyright-statement xml:lang="ru">Copyright ©; 2024, Эко-Вектор</copyright-statement><copyright-statement xml:lang="zh">Copyright ©; 2024,</copyright-statement><copyright-year>2024</copyright-year><copyright-holder xml:lang="en">Eco-Vector</copyright-holder><copyright-holder xml:lang="ru">Эко-Вектор</copyright-holder><ali:free_to_read xmlns:ali="http://www.niso.org/schemas/ali/1.0/" start_date="2027-12-28"/><license><ali:license_ref xmlns:ali="http://www.niso.org/schemas/ali/1.0/">https://creativecommons.org/licenses/by-nc-nd/4.0/</ali:license_ref></license></permissions><self-uri xlink:href="https://j-morphology.com/1026-3543/article/view/642031">https://j-morphology.com/1026-3543/article/view/642031</self-uri><abstract xml:lang="en"><p><bold>BACKGROUND:</bold><bold> </bold>Eosinophilic granulocytes exhibit antibacterial, antiparasitic, and antiviral activity and play a significant role in the pathogenesis of diseases such as eosinophilic esophagitis and colitis. However, standard methods for their identification have significant limitations, which complicate precise quantification and diagnosis of associated conditions.</p> <p><bold>AIM:</bold><bold><italic> </italic></bold>To develop and evaluate a novel staining method for accurate and selective identification of eosinophilic granulocytes in the gastrointestinal tract tissues.</p> <p><bold>METHODS:</bold><bold><italic> </italic></bold>The study was conducted using histological sections of the stomach, small intestine, and large intestine from male Wistar rats (<italic>n</italic> = 16). Tissue sections were stained with a combination of eosin and methylene green.</p> <p><bold>RESULTS:</bold><bold><italic> </italic></bold>The developed method provided high-contrast staining of eosinophilic granulocytes and demonstrated greater accuracy compared to conventional hematoxylin and eosin staining. Eosinophils were distinctly visible on slides stained with the new method even under low magnification. The technique enabled easy identification, enumeration, and localization of eosinophilic granulocytes within the gastrointestinal tract tissues.</p> <p><bold>CONCLUSION:</bold><bold><italic> </italic></bold>The proposed staining method allows for precise and selective detection of eosinophilic granulocytes in the gastrointestinal mucosa of rats and shows promise for use in diagnosing conditions associated with eosinophilic infiltration.</p></abstract><trans-abstract xml:lang="ru"><p><bold>Обоснование.</bold><bold> </bold>Эозинофильные гранулоциты проявляют антибактериальную, антипаразитарную и противовирусную активность, а также являются важными участниками патогенеза таких заболеваний, как эозинофильный эзофагит и колит. Однако стандартные методы их выявления имеют существенные ограничения, которые затрудняют количественное определение эозинофилов и диагностику ряда заболеваний.</p> <p><bold>Цель</bold><bold><italic> </italic></bold>— разработка и оценка нового метода окрашивания для точного и селективного выявления эозинофильных гранулоцитов в тканях желудочно-кишечного тракта.</p> <p><bold>Методы.</bold> Исследование проведено на срезах желудка, тонкой и толстой кишки самцов крыс линии Вистар (<italic>n</italic>=16). Гистологические срезы тканей окрашивали эозином и метиленовым зелёным.</p> <p><bold>Результаты.</bold><bold> </bold>Разработанный метод обеспечивает высокую контрастность окрашивания эозинофильных гранулоцитов и превосходит традиционный метод окраски гематоксилином и эозином по точности. Эозинофильные гранулоциты на препаратах, окрашенных новым методом, чётко различимы и хорошо выявляются даже при малых увеличениях объектива. Предложенный метод позволяет легко идентифицировать эозинофилы, проводить их подсчёт и определять локализацию в тканях желудочно-кишечного тракта.</p> <p><bold>Заключение.</bold><bold> </bold>Новый метод позволяет точно и селективно выявлять эозинофильные гранулоциты в слизистой оболочке пищеварительного тракта крыс, что делает его перспективным для возможного применения в диагностике заболеваний, связанных с эозинофильной инфильтрацией.</p></trans-abstract><trans-abstract xml:lang="zh"><p>论证嗜酸性粒细胞具有抗菌、抗寄生虫和抗病毒作用，并在嗜酸性食管炎、嗜酸性结肠炎等疾病的发病机制中发挥关键作用。嗜酸性粒细胞具有抗菌、抗寄生虫和抗病毒作用，并在嗜酸性食管炎和嗜酸性结肠炎的发病机制中发挥重要作用。</p> <p>目的。开发并评估一种新的染色方法，用于胃肠道组织中嗜酸性粒细胞的精准且选择性识别。</p> <p>材料与方法。本研究在Wistar系雄性大鼠（n=16）的胃、小肠和大肠组织切片上进行。组织切片采用联合伊红与亚甲基绿的染色方法处理。</p> <p>结果。所建立的染色方法显著提高了嗜酸性粒细胞的染色对比度，其识别准确性优于传统的苏木精-伊红染色法。在采用该新型染色方法处理的切片中，嗜酸性粒细胞在低倍镜下即可清晰可见，易于识别。该方法可有效识别嗜酸性粒细胞，便于计数并确定其在胃肠组织中的定位。</p> <p>结论。该新染色方法能够在大鼠消化道黏膜中实现对嗜酸性粒细胞的精准、选择性识别，具备作为诊断嗜酸性粒细胞浸润相关疾病工具的应用前景。</p></trans-abstract><kwd-group xml:lang="en"><kwd>eosinophils</kwd><kwd>gastrointestinal tract</kwd><kwd>hematoxylin and eosin staining</kwd><kwd>rats</kwd></kwd-group><kwd-group xml:lang="ru"><kwd>эозинофилы</kwd><kwd>желудочно-кишечный тракт</kwd><kwd>гематоксилин и эозин</kwd><kwd>крысы</kwd></kwd-group><kwd-group xml:lang="zh"><kwd>嗜酸性粒细胞</kwd><kwd>胃肠道</kwd><kwd>苏木精-伊红染色</kwd><kwd>大鼠</kwd></kwd-group><funding-group><award-group><funding-source><institution-wrap><institution xml:lang="ru">Министерство науки и высшего образования Российской Федерации</institution></institution-wrap><institution-wrap><institution xml:lang="en">Ministry of Science and Higher Education of the Russian Federation</institution></institution-wrap><institution-wrap><institution xml:lang="zh">俄羅斯聯邦科學與高等教育部</institution></institution-wrap></funding-source><award-id>075-03-2023-657/4</award-id></award-group></funding-group></article-meta></front><body></body><back><ref-list><ref id="B1"><label>1.</label><mixed-citation>Rothenberg ME, Hogan SP. 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