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Cell Biology JoVE (Open Access) Citable · DOI

A Non-invasive Way to Isolate and Phenotype Cells from the Conjunctiva

DOI: 10.3791/55591-v
What you'll learn
  • ✓ Perform non-invasive impression cytology on human conjunctival tissue
  • ✓ Prepare and analyze conjunctival cells using flow cytometry
  • ✓ Identify and phenotype immune cell subsets in ocular surface epithelium
Protocol

Biopharma Insights The exposed normal ocular surface consists of cornea and conjunctiva. Epithelial cells, goblet cells and immune cells are present in the conjunctiva. Here, a non-invasive, technique of impression cytology is described using an impression cytology device and flow cytometry to analyze immune cells in the conjunctiva.

Difficulty
intermediate
Total time
~45 min per subject (collection + flow cytometry analysis)
Model organism
Human conjunctival epithelium
Biosafety
BSL-1

Steps

1
Collect conjunctival cells using impression cytology device

Apply an impression cytology device to the exposed conjunctival surface of the eye to non-invasively harvest epithelial cells, goblet cells, and immune cells.

▶ 01:14
2
Prepare cells and perform flow cytometric phenotyping

Process collected conjunctival cells and use flow cytometry to identify and characterize immune cell populations and subsets present in the sample.

▶ 05:48
3
Analyze representative T cell subset distributions

Examine flow cytometry data to identify T cell populations and immune phenotypes characteristic of healthy human ocular surface tissue.

▶ 06:35
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