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Protocol for isolating and characterizing human vitreous immune cell infiltrates by flow cytometry and single-cell
Sandhya Rani Bhanja1, Sayantan Ghosh1, Pankaj Kumar2
1Ocular Immunology Laboratory, Prof Brien Holden Eye Research Centre, LV Prasad Eye Institute, Kallam Anji Reddy Campus, Hyderabad, India.
STAR Protocols
|May 18, 2025
Summary
This study details a new protocol for isolating immune cells from eye vitreous. This method allows for detailed analysis of intraocular immune responses without eye damage.
Area of Science:
- Ophthalmology
- Immunology
- Molecular Biology
Background:
- Immune cells in the eye's vitreous humor offer insights into intraocular immune responses.
- Accessing these cells without harming the eye is crucial for research.
Purpose of the Study:
- To present a customized protocol for isolating and characterizing immune cell infiltrates from human vitreous samples.
- To optimize immune cell yield for downstream analyses.
Main Methods:
- Surgical modifications for enhanced vitreous sample collection.
- Flow cytometry for immune cell characterization.
- Single-cell transcriptomics for in-depth analysis.
Main Results:
- The protocol effectively isolates and characterizes vitreous immune cells.
- Maximized immune cell yield is achieved through specific measures.
- The method preserves the eye's structural and functional integrity.
Conclusions:
- The developed protocol provides a valuable tool for studying intraocular immunity.
- This method enables detailed analysis of immune cell infiltrates in various ocular conditions.
- Further research can utilize this protocol to understand immune-mediated eye diseases.
