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Label-free quantitative imaging of conjunctival goblet cells.
Noseong Park1, Suil Jeon2, Seonghan Kim2
1Division of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-gu, Pohang, Gyeongbuk, 37673, Republic of Korea.
The Ocular Surface
|January 25, 2025
Summary
Quantitative oblique back-illumination microscopy (qOBM) offers high-contrast, label-free visualization of conjunctival goblet cells (GCs). This technique successfully assesses GC function and changes, showing promise for ocular surface disease diagnostics.
Area of Science:
- Ophthalmology
- Microscopy
- Cell Biology
Background:
- Conjunctival goblet cells (GCs) are crucial for ocular surface health.
- Assessing GC function is vital for diagnosing and managing ocular surface diseases.
- Existing imaging techniques may lack the contrast or label-free capabilities for detailed GC analysis.
Purpose of the Study:
- Introduce and validate quantitative oblique back-illumination microscopy (qOBM) as a novel imaging technique.
- Demonstrate qOBM's ability to visualize conjunctival goblet cells (GCs) without labels.
- Assess the functional changes of GCs using qOBM.
Main Methods:
- Developed qOBM and validated it against moxifloxacin-based fluorescence microscopy (MBFM) and PAS staining.
- Tested qOBM on polystyrene beads for initial calibration and accuracy.
- Performed longitudinal ex-vivo and in-vivo imaging of mouse conjunctiva under hyperosmotic stress and normal conditions.
Main Results:
- qOBM accurately imaged beads, aligning with theoretical phase delay predictions.
- High-contrast visualization of GCs was achieved, with an average phase delay of 0.59 ± 0.25 radians in normal mouse conjunctiva.
- Hyperosmotic stress significantly reduced GC phase delays, consistent with MBFM and PAS staining, indicating functional changes.
Conclusions:
- qOBM is a validated, high-contrast, label-free imaging method for GCs.
- The technique enables effective functional assessment of GCs.
- qOBM shows significant potential for advancing ocular surface disease research and clinical diagnostics.
