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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.
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http://dx.doi.org/10.1016/j.xpro.2024.103444 | DOI Listing |
Nat Immunol
September 2025
Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO, USA.
Crohn's disease pathology is modeled in TNF mice that overproduce tumor necrosis factor (TNF) to drive disease through TNF receptors. An alternative ligand for TNF receptors, soluble LTα, is produced by B cells, but has received scarce attention because LTα also partners with LTβ to generate membrane-tethered LTαβ that promotes tertiary lymphoid tissue-another feature of Crohn's disease. We hypothesized that B cell-derived LTαβ would critically affect ileitis in TNF mice.
View Article and Find Full Text PDFPigment Cell Melanoma Res
September 2025
Department of Dermatology, Beijing Tongren Hospital, Capital Medical University, Beijing, China.
The diagnostic approaches for Hermansky-Pudlak Syndrome (HPS) include genetic sequencing, immunoblotting, electron microscopy (EM), and flow cytometry with mepacrine staining. However, these methods are often impractical for routine clinical use due to high cost, technical complexity, and limited availability. In this study, we evaluated dense granules (DGs) function in HPS mouse models using flow cytometry with mepacrine and FluoZin-3 staining.
View Article and Find Full Text PDFJ Neurosci Methods
September 2025
The Schepens Eye Research Institute of Massachusetts Eye and Ear, Boston, MA 02114, United States; Department of Ophthalmology, Harvard Medical School, Boston, MA 02114, United States; Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53706, United States. Electronic
Background: Gene and cell therapies hold promise for restoring vision in hereditary and advanced optic neuropathies. Accurate evaluation of these therapies requires advanced imaging methods that can visualize transplanted cells within intact retinal tissue.
New Method: We present a whole-mount tissue-clearing workflow optimized for the mouse retina and optic nerve to improve visualization of donor neuron integration following cell transplantation.
Bio Protoc
August 2025
The Scripps Research Institute, Neuroscience Department and Dorris Neuroscience Center, La Jolla, CA, USA.
The process of moving proteins and organelles along the axon is essential for neuronal survival and function, ensuring proper communication between the cell body and distant synapses. The efficient and precise delivery of proteins via axon transport is critical for processes ranging from synaptic plasticity and neurotransmission to neuronal growth and maintenance. However, the identities of all the transported proteins have only recently begun to be investigated.
View Article and Find Full Text PDFMethods Mol Biol
August 2025
Department of Physiology & Neurobiology, University of Connecticut, Storrs, CT, USA.
The mouse retina is a widely used model to study sprouting angiogenesis. Most blood vessel staining protocols focus on the superficial vascular layer. Here, we describe a method to obtain thick sections of retina whole-mounts that allow for detailed visualization of diving vascular sprouts, and deep and intermediate-layer angiogenesis.
View Article and Find Full Text PDF