利用双重染色和可视化技术定量分析全血中的吞噬作用

IF 2.3 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Elena V. Lysakova, Alexander N. Shumeev, Sergei A. Chuvpilo, Viktor S. Laktyushkin, Natalia A. Arsentieva, Mikhail Yu. Bobrov, Stanislav A. Rybtsov
{"title":"利用双重染色和可视化技术定量分析全血中的吞噬作用","authors":"Elena V. Lysakova,&nbsp;Alexander N. Shumeev,&nbsp;Sergei A. Chuvpilo,&nbsp;Viktor S. Laktyushkin,&nbsp;Natalia A. Arsentieva,&nbsp;Mikhail Yu. Bobrov,&nbsp;Stanislav A. Rybtsov","doi":"10.1134/S0006297924050122","DOIUrl":null,"url":null,"abstract":"<p>Phagocytosis is an essential innate immunity function in humans and animals. A decrease in the ability to phagocytize is associated with many diseases and aging of the immune system. Assessment of phagocytosis dynamics requires quantification of bacteria inside and outside the phagocyte. Although flow cytometry is the most common method for assessing phagocytosis, it does not include visualization and direct quantification of location of bacteria. Here, we used double-labeled <i>Escherichia coli</i> cells to evaluate phagocytosis by flow cytometry (cell sorting) and confocal microscopy, as well as employed image cytometry to provide high-throughput quantitative and spatial recognition of the double-labeled <i>E. coli</i> associated with the phagocytes. Retention of pathogens on the surface of myeloid and lymphoid cells without their internalization was suggested to be an auxiliary function of innate immunity in the fight against infections. The developed method of bacterial labeling significantly increased the accuracy of spatial and quantitative measurement of phagocytosis in whole blood and can be recommended as a tool for phagocytosis assessment by image cytometry.</p>","PeriodicalId":483,"journal":{"name":"Biochemistry (Moscow)","volume":"89 5","pages":"923 - 932"},"PeriodicalIF":2.3000,"publicationDate":"2024-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Quantitative Analysis of Phagocytosis in Whole Blood Using Double Staining and Visualization\",\"authors\":\"Elena V. Lysakova,&nbsp;Alexander N. Shumeev,&nbsp;Sergei A. Chuvpilo,&nbsp;Viktor S. Laktyushkin,&nbsp;Natalia A. Arsentieva,&nbsp;Mikhail Yu. Bobrov,&nbsp;Stanislav A. Rybtsov\",\"doi\":\"10.1134/S0006297924050122\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Phagocytosis is an essential innate immunity function in humans and animals. A decrease in the ability to phagocytize is associated with many diseases and aging of the immune system. Assessment of phagocytosis dynamics requires quantification of bacteria inside and outside the phagocyte. Although flow cytometry is the most common method for assessing phagocytosis, it does not include visualization and direct quantification of location of bacteria. Here, we used double-labeled <i>Escherichia coli</i> cells to evaluate phagocytosis by flow cytometry (cell sorting) and confocal microscopy, as well as employed image cytometry to provide high-throughput quantitative and spatial recognition of the double-labeled <i>E. coli</i> associated with the phagocytes. Retention of pathogens on the surface of myeloid and lymphoid cells without their internalization was suggested to be an auxiliary function of innate immunity in the fight against infections. The developed method of bacterial labeling significantly increased the accuracy of spatial and quantitative measurement of phagocytosis in whole blood and can be recommended as a tool for phagocytosis assessment by image cytometry.</p>\",\"PeriodicalId\":483,\"journal\":{\"name\":\"Biochemistry (Moscow)\",\"volume\":\"89 5\",\"pages\":\"923 - 932\"},\"PeriodicalIF\":2.3000,\"publicationDate\":\"2024-05-31\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biochemistry (Moscow)\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://link.springer.com/article/10.1134/S0006297924050122\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochemistry (Moscow)","FirstCategoryId":"99","ListUrlMain":"https://link.springer.com/article/10.1134/S0006297924050122","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

摘要 吞噬是人类和动物的一项基本先天免疫功能。吞噬能力的下降与许多疾病和免疫系统的衰老有关。评估吞噬动态需要对吞噬细胞内外的细菌进行量化。虽然流式细胞术是评估吞噬能力最常用的方法,但它不包括细菌位置的可视化和直接量化。在这里,我们使用双标记大肠杆菌细胞,通过流式细胞术(细胞分拣)和共聚焦显微镜评估吞噬作用,并采用图像细胞术对与吞噬细胞相关的双标记大肠杆菌进行高通量定量和空间识别。病原体滞留在髓细胞和淋巴细胞表面而不被内化被认为是先天性免疫在抗感染中的一种辅助功能。所开发的细菌标记方法大大提高了对全血中吞噬作用进行空间和定量测量的准确性,可推荐用作图像细胞术评估吞噬作用的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Quantitative Analysis of Phagocytosis in Whole Blood Using Double Staining and Visualization

Quantitative Analysis of Phagocytosis in Whole Blood Using Double Staining and Visualization

Phagocytosis is an essential innate immunity function in humans and animals. A decrease in the ability to phagocytize is associated with many diseases and aging of the immune system. Assessment of phagocytosis dynamics requires quantification of bacteria inside and outside the phagocyte. Although flow cytometry is the most common method for assessing phagocytosis, it does not include visualization and direct quantification of location of bacteria. Here, we used double-labeled Escherichia coli cells to evaluate phagocytosis by flow cytometry (cell sorting) and confocal microscopy, as well as employed image cytometry to provide high-throughput quantitative and spatial recognition of the double-labeled E. coli associated with the phagocytes. Retention of pathogens on the surface of myeloid and lymphoid cells without their internalization was suggested to be an auxiliary function of innate immunity in the fight against infections. The developed method of bacterial labeling significantly increased the accuracy of spatial and quantitative measurement of phagocytosis in whole blood and can be recommended as a tool for phagocytosis assessment by image cytometry.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Biochemistry (Moscow)
Biochemistry (Moscow) 生物-生化与分子生物学
CiteScore
4.70
自引率
3.60%
发文量
139
审稿时长
2 months
期刊介绍: Biochemistry (Moscow) is the journal that includes research papers in all fields of biochemistry as well as biochemical aspects of molecular biology, bioorganic chemistry, microbiology, immunology, physiology, and biomedical sciences. Coverage also extends to new experimental methods in biochemistry, theoretical contributions of biochemical importance, reviews of contemporary biochemical topics, and mini-reviews (News in Biochemistry).
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信