A Protocol for Immunostaining of Macrophages in Whole-Mount Mouse Cornea, Conjunctiva, and Lacrimal Gland

Authors

Saleh Alfuraih, Chapman UniversityFollow
Dhruv Patel, Chapman University
Wonsuk Choi, Chapman UniversityFollow
Karthikeyan Ramasamy, Chapman University
Rais Ansari, Nova Southeastern University
Ajay Sharma, Chapman UniversityFollow

Document Type

Article

Publication Date

11-7-2024

Abstract

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.

Comments

This article was originally published in STAR Protocols, volume 5, issue 4, in 2024. https://doi.org/10.1016/j.xpro.2024.103444

Recommended Citation

Alfuraih S, Patel D, Choi W, Ramasamy K, Ansari R, Sharma A. A protocol for immunostaining of macrophages in whole-mount mouse cornea, conjunctiva, and lacrimal gland. STAR Protocols. 2024;5(4):103444. https://doi.org/10.1016/j.xpro.2024.103444

Copyright

The authors

Creative Commons License

This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.