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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Bibliographic Details
Published in:STAR Protocols
Main Authors: Saleh Alfuraih, Dhruv Patel, Wonsuk Choi, Karthikeyan Ramasamy, Rais Ansari, Ajay Sharma
Format: Article
Language:English
Published: Elsevier 2024-12-01
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724006099
Description
Summary: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.
ISSN:2666-1667