Protocol to examine immune subpopulations in murine conjunctiva and lacrimal gland using flow cytometry

Area of Science:

• Immunology
• Ophthalmology
• Cell Biology

Background:

• The ocular surface, including the conjunctiva and lacrimal gland, harbors a unique immune microenvironment.
• Understanding immune cell populations in these tissues is crucial for diagnosing and treating ocular surface diseases.

Purpose of the Study:

• To establish a detailed and reproducible protocol for the flow cytometric analysis of immune cells within the murine conjunctiva and lacrimal gland.
• To provide a standardized method for researchers investigating ocular immunology.

Main Methods:

• Dissection of the murine conjunctiva and lacrimal gland.
• Preparation of high-quality single-cell suspensions from ocular tissues.
• Application of comprehensive antibody staining panels for immune cell identification.
• Optimization of flow cytometry voltage settings, compensation adjustments, and gating strategies.

Main Results:

• The protocol yields high-quality single-cell suspensions suitable for flow cytometry.
• Comprehensive staining panels allow for detailed characterization of immune cell subsets.
• Optimized gating strategies ensure accurate identification and quantification of immune cells.

Conclusions:

• This protocol provides a robust framework for the examination of immune cells in the murine conjunctiva and lacrimal gland.
• The standardized methodology facilitates consistent and comparable results in ocular immunology research.
• This method aids in advancing the understanding of ocular immune responses and related pathologies.