A protocol for immunostaining of macrophages in whole-mount mouse cornea, conjunctiva, and lacrimal gland
Summary: The healthy lacrimal functional unit contains a resident macrophage population. Here, we present a protocol for immunofluorescent staining of macrophage markers, CD11b, F4/80, and CD206, in whole-mount mouse cornea, conjunctiva, and 50-μM-thick lacrimal gland section. We describe steps for...
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Elsevier
2024-12-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724006099 |
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| author | Saleh Alfuraih Dhruv Patel Wonsuk Choi Karthikeyan Ramasamy Rais Ansari Ajay Sharma |
| author_facet | Saleh Alfuraih Dhruv Patel Wonsuk Choi Karthikeyan Ramasamy Rais Ansari Ajay Sharma |
| author_sort | Saleh Alfuraih |
| collection | DOAJ |
| description | Summary: The healthy lacrimal functional unit contains a resident macrophage population. Here, we present a protocol for immunofluorescent staining of macrophage markers, CD11b, F4/80, and CD206, in whole-mount mouse cornea, conjunctiva, and 50-μM-thick lacrimal gland section. We describe steps for dissection, fixation, permeabilization, and blocking. We then detail procedures for the detection and spatial localization of macrophages through immunostaining and confocal imaging. This approach circumvents the need to obtain thin tissue sections and acquire macrophage images from each tissue section. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-d3aaa58d808a4022827d4e9ece80af82 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-d3aaa58d808a4022827d4e9ece80af822024-11-20T05:08:14ZengElsevierSTAR Protocols2666-16672024-12-0154103444A protocol for immunostaining of macrophages in whole-mount mouse cornea, conjunctiva, and lacrimal glandSaleh Alfuraih0Dhruv Patel1Wonsuk Choi2Karthikeyan Ramasamy3Rais Ansari4Ajay Sharma5Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Chapman University, Irvine, CA 92618, USA; Department of Pharmaceutical Sciences, Barry and Judy Silverman College of Pharmacy, Health Professions Division, Nova Southeastern University, Fort Lauderdale, FL, USA; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Northern Border University, Rafha, Saudi ArabiaDepartment of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Chapman University, Irvine, CA 92618, USADepartment of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Chapman University, Irvine, CA 92618, USADepartment of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Chapman University, Irvine, CA 92618, USADepartment of Pharmaceutical Sciences, Barry and Judy Silverman College of Pharmacy, Health Professions Division, Nova Southeastern University, Fort Lauderdale, FL, USADepartment of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Chapman University, Irvine, CA 92618, USA; Corresponding authorSummary: The healthy lacrimal functional unit contains a resident macrophage population. Here, we present a protocol for immunofluorescent staining of macrophage markers, CD11b, F4/80, and CD206, in whole-mount mouse cornea, conjunctiva, and 50-μM-thick lacrimal gland section. We describe steps for dissection, fixation, permeabilization, and blocking. We then detail procedures for the detection and spatial localization of macrophages through immunostaining and confocal imaging. This approach circumvents the need to obtain thin tissue sections and acquire macrophage images from each tissue section. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724006099immunologymicroscopysystems biology |
| spellingShingle | Saleh Alfuraih Dhruv Patel Wonsuk Choi Karthikeyan Ramasamy Rais Ansari Ajay Sharma A protocol for immunostaining of macrophages in whole-mount mouse cornea, conjunctiva, and lacrimal gland STAR Protocols immunology microscopy systems biology |
| title | A protocol for immunostaining of macrophages in whole-mount mouse cornea, conjunctiva, and lacrimal gland |
| title_full | A protocol for immunostaining of macrophages in whole-mount mouse cornea, conjunctiva, and lacrimal gland |
| title_fullStr | A protocol for immunostaining of macrophages in whole-mount mouse cornea, conjunctiva, and lacrimal gland |
| title_full_unstemmed | A protocol for immunostaining of macrophages in whole-mount mouse cornea, conjunctiva, and lacrimal gland |
| title_short | A protocol for immunostaining of macrophages in whole-mount mouse cornea, conjunctiva, and lacrimal gland |
| title_sort | protocol for immunostaining of macrophages in whole mount mouse cornea conjunctiva and lacrimal gland |
| topic | immunology microscopy systems biology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724006099 |
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